Cell. Mol. Life Sci.

65 (2008) 2001 – 2018

1420-682X/08/132001-18 Cellular and Molecular Life Sciences
DOI 10.1007/s00018-008-8001-x

Birkhuser Verlag, Basel, 2008

Review

Melatonin, hormone of darkness and more – occurrence, control

mechanisms, actions and bioactive metabolites
R. Hardeland

Johann Friedrich Blumenbach Institute of Zoology and Anthropology, University of Gçttingen, Berliner Str. 28,
37073 Gçttingen (Germany), Fax: + 49 551 395438, e-mail: rhardel@gwdg.de

Received 2 January; received after revision 6 February 2008; accepted 8 February 2008
Online First 17 March 2008

Abstract. In its role as a pineal hormone, melatonin is levels of gene expression and/or enzyme stability
a pleiotropic, nocturnally peaking and systemically influenced by phosphorylation and interaction with
acting chronobiotic. These effects are largely ex- 14-3-3 proteins. Melatonin is not only a hormone but is
plained by actions via G protein-coupled membrane also synthesized in numerous extrapineal sites, in
receptors found in the suprachiasmatic nucleus, but which it sometimes attains much higher quantities
also in numerous other sites. Nuclear (ROR/RZR), than in the pineal and the circulation. It is also present
cytoplasmic (quinone reductase-2, calmodulin, calre- in many taxonomically distant groups of organisms,
ticulin) and mitochondrial binding sites and radical- including bacteria, fungi, and plants. Moreover, mel-
scavenging properties contribute to the actions of atonin is a source of bioactive metabolites, such as 5-
melatonin. Regulation of pineal melatonin biosyn- methoxytryptamine, N1-acetyl-N2-formyl-5-methoxy-
thesis is largely explained by control mechanisms kynuramine and N1-acetyl-5-methoxykynuramine.
acting on arylalkylamine N-acetyltransferase, at the

Keywords. AFMK, AMK, cinnoline, indoleamine, melatonin-binding site, 5-methoxytryptamine, nitric oxide,
pineal gland.

Introduction and the circulation with prominent nocturnal peaks,

which were suppressed by light [3 – 7]. The chrono-
Melatonin was discovered as a pineal hormone, which biological roles of melatonin as a mediator of dark
lightens skin of fish and amphibia by concentrating signals regulating both circadian oscillators and sea-
mobile melanosomes of melanocytes (melanophores) sonality have been reviewed many times and would
involved in physiological color change [1]. Even this exceed the frame of this article.
frequently cited observation from a pioneering work is Meanwhile, melatonin has turned out to be a ubiq-
not without exception. In pencil fish, melanocytes uitous compound, detected in all major taxa studied so
showed area-specific melanosome concentration or far, including bacteria, dinoflagellates and other
dispersion [2]. Therefore, melatonin is not just anoth- eukaryotic protists, macroalgae, plants, fungi, and
er example of the various skin-lightening hormones various groups of invertebrate animals [for details see
present in several animal taxa, but rather transmits refs 8 – 10]. In brief, the following lessons can be
signals eliciting different site-specific responses. This deduced from these findings: (i) melatonin is not
view was confirmed by the discovery of high-ampli- generally associated with darkness, but can sometimes
tude circadian melatonin rhythms in the pineal gland also be elevated during photophase; (ii) in some
2002 R. Hardeland Melatonin, hormone and more

species, it may even be arrhythmic; (iii) where it is retina and parietal organ, also being extrusions of the
found to exhibit circadian oscillations, these are not intermediate brain, may appear as variants of the
necessarily implicated in seasonality; (iv) in plants, same theme known from the pineal gland. They can
morphogenetic and photoprotective actions may be of likewise generate robust, nocturnally peaking circa-
importance; (v) the concentrations of non-vertebrate dian rhythms. However, with the exception of am-
melatonin can vary extremely between species or taxa phibian and some avian retinas, melatonin is only
and sometimes attain values in the upper micromolar poorly released from these organs. Consequently,
range. Therefore, melatonin can no longer be seen melatonin serves additional functions and is not only
solely as a trace compound, nor generally as a acting as a hormone. Retinal melatonin efficiently
hormone, and its functions have to be highly divergent downregulates dopamine formation and release [11,
in the various organisms. 13]. Despite the similar rhythmicities of pineal and
Although this indoleamine has been perceived for retinal melatonin, their regulation can be entirely
decades in a mainly chronobiological context, even different. While melatonin in the mammalian pineal is
vertebrate melatonin is not exclusively a hormone largely regulated by norepinephrine of sympathetic
present in only minute quantities and acting via the G origin, retinal melatonin biosynthesis is stimulated via
protein-coupled membrane receptors MT1 and MT2. GABAA and, partially, GABAB receptors [13]. The
High-amplitude circadian oscillations, which mediate light-dependent decrease of retinal melatonin is
the signal darkness, are not generally found. Tissue caused by dopamine via D1 or D4 receptors [18].
melatonin enters the circulation poorly is and thus Differences in metabolism exist, too, because retinal
only partially removed by hepatic first-pass metabo- melatonin is largely degraded by deacetylation to 5-
lism and not quantitatively excreted as 6-sulfatoxy- methoxytryptamine (5-MT) [12, 19], a compound
melatonin. Therefore, the role as a mediator of the which can be further converted to 5-methoxytrypto-
signal darkness only reflects a part of the wider phol (5-ML) or 5-methoxyindoleacetic acid (5-
spectrum of actions, presence, dynamics and metab- MIAA) [8, 10, 20]. In the retina, deacetylation to 5-
olism exhibited by this remarkably pleiotropic and MT can be catalyzed by a specific melatonin deace-
ubiquitous indoleamine. This review will deal with tylase, but in other organs or organisms by less specific
both the considerable progress made in understanding aryl acylamidases (AAAs) or eserine-sensitive ace-
the cell biology of the classic functions of melatonin tylcholinesterase [20]. 5-MT is a bioactive compound
and its roles beyond them. too [8, 10, 20] and may play a role of its own in the
retina. In cultured retinal cells, it reportedly prevented
the forskolin-induced rise in cyclic AMP, independ-
Extrapineal melatonin: remarkable quantities and ently of the melatonin receptors [21]. Where not
non-classic roles released from an organ, melatonin cannot not be
degraded in the liver or appear as urinary 6-sulfatox-
Extrapineal melatonin is somehow reminiscent of ymelatonin. In the brain, substituted kynuramines
cosmological dark matter. There is by far more were identified as major metabolites [22].
melatonin outside than inside the pineal gland and While robust circadian rhythmicity is typical for
circulation, but its extrapineal functions are poorly pineal, parietal and retina, this is not so with the
understood, whereas the hormone secreted by the rodent Harderian gland. Diurnal changes are poorly
pineal gland and its transmission of dark signals is part expressed or almost absent, except for a transient drop
of general knowledge and awareness. The much after onset of light [14, 16]. Therefore, melatonin is not
higher quantities of extrapineal melatonin are fre- always associated with darkness in vertebrates. Var-
quently not perceived by researchers or are consid- iations of Harderian melatonin were observed within
ered irrelevant. However, extrapineal melatonin will sexual or seasonal cycles, but these are related to
substantially widen our understanding of the numer- changes in gonadosteroids [14, 23]. Thus, annual
ous roles of this molecule. fluctuations appear as a consequence rather than a
Pineal melatonin is formed from serotonin by two cause of reproductive cycles.
steps catalyzed by arylalkylamine N-acetyltransferase Melatonin biosynthesis has been reported, or as-
(AA-NAT) and hydroxyindole O-methyltransferase sumed, for various additional tissues and cells, and, in
(HIOMT) [6, 7]. In extrapineal sites and outside most cases, a transmission of dark signals seems highly
vertebrates, other enzymes can be involved and unlikely or impossible. Circadian rhythms, if present,
regulation mechanisms may be different. Extrapineal frequently exhibit small amplitudes or are at the
melatonin synthesis has been identified, e,g. in the borderline of detection. In the mammalian brain,
retina [11 – 13], the Harderian gland [14 – 16] and later other sites of melatonin formation may exist, although
in the parietal organ of reptiles [17]. At first glance, the evidence is relatively weak to date, whereas more
Cell. Mol. Life Sci. Vol. 65, 2008 Review Article 2003

information exists on the synthesis of the precursor, N- 30 mM melatonin, under consideration of an erratum)
acetylserotonin (NAS). O-methylation of NAS is not [36]. A clarification of this discrepancy is urgently
necessarily an action of HIOMT, as in pineal, retina required, to decide between dysregulation in cultured,
and Harderian gland, but can be alternately catalyzed immortalized cells and inefficiency of safe extraction
by less specific methyltransferases. Relatively high from biopsies, as caused by oxidants in biological
concentrations (up to 0.7 mM) of melatonin in the material [9, 20].
brain have been reported [24], a finding which would Melatonin formation has been reported for several
be in favor of local melatonin synthesis in some areas. other vertebrate tissues and cells, such as bone marrow
Abundance and sites of formation of 5-hydroxylated [37] – along with assumptions concerning hemato-
and 5-methoxylated indoles in the central nervous poietic activity of melatonin [38] – and various types of
system have been recently summarized [20]. leukocytes [39, 40], platelets, erythrocytes and the
Gastrointestinal melatonin has been reviewed several membranous cochlea [summarized in ref. 39]. Ele-
times [e.g., 25 – 27]. The indoleamine is produced in vated concentrations in other tissues [40, 41] may be
enterochromaffin cells, but circulating melatonin can explained by uptake from the circulation. In some
be additionally taken up from the blood. Although cases, melatonin formation may result from inevitable
concentrations remain moderate, the entire gastro- side reactions by unspecific N-acetyl and O-methyl-
intestinal tract contains, owing to its size, about 400 – transferases, e.g., in erythrocytes.
500 times more melatonin than the pineal gland [26,
27]. Its fate is complex, because the gut acts both as
source and sink. Uptake of circulating melatonin has Regulation of melatonin biosynthesis
been studied by elevating its plasma level during
daytime to nighttime values [28]. Substantial amounts The classic pathway of melatonin formation involves
were subsequently found unmetabolized in the intes- four steps, starting with tryptophan 5-hydroxylase,
tinal lumen [28]. This is in good agreement with high followed by 5-hydroxytryptophan decarboxylation by
quantities of melatonin in the bile fluid [29] and aromatic amino acid decarboxylase, N-acetylation of
enterohepatic cycling [29, 30]. Additionally, melato- serotonin by AA-NAT, and O-methylation of NAS by
nin is taken up from food. Diurnal plasma levels were HIOMT. Alternately, homologs or structurally differ-
increased by natural diets rich in melatonin, or ent N-acetyl- and O-methyltransferases can be in-
decreased by melatonin-depleted food [27, 31]. A volved. In principle, the steps could also take place in
contribution of intestinal bacteria to melatonin in the different sequential orders, e.g., serotonin ! 5-
gut seems possible, but requires further investigation MT ! melatonin, or 5-hydroxytryptophan ! 5-me-
[9]. Gastrointestinal melatonin can be released to the thoxytryptophan ! 5-MT ! melatonin, but, in all
circulation, in terms of a post-prandial response [26, cases studied so far, the alternate sequences were of
27], in particular, by elevated tryptophan [32]. Mel- minor importance. In organs or organisms which
atonin surges elicited by this amino acid are consid- deacetylate melatonin [42], re-acetylation of 5-MT
erable in height, but of relatively short duration. may be of some relevance [10]. In the classic pathway,
During daytime, they remain chronobiologically al- AA-NAT is usually believed to be the rate-limiting
most ineffective, since they appear in the silent zone of enzyme, although tryptophan 5-hydroxylase can rep-
the circadian phase-response curve [27]. Gastrointes- resent an additional control step, e.g., in amphibia.
tinal melatonin can also exhibit a circadian rhythmic- Exceptions to this rule have been described. In
ity of relatively low amplitude (maximum/minimum Drosophila, NAS concentrations are three orders of

2 : 1) [27], which is sometimes barely detectable [26, magnitude higher than those of melatonin [43], and
27]. thus O-methylation is limiting, a finding explained by
Mammalian skin has been reported to be another site the loss of the HIOMT gene, so that methyl transfer
of melatonin biosynthesis. The formation and pres- has to be catalyzed by other, presumably less specific
ence of the enzymes required have been demonstrated enzymes [20]. A functionally similar situation seems
[33]. Serotonin N-acetylation is catalyzed by either to be present in the retinas of primates and ungulates,
AA-NAT or by an arylamine N-acetyltransferase in which HIOMT is not or poorly expressed at both
subform [34]. An important and obviously unsettled mRNA and protein levels [7]. Whether this leads to a
point concerns melatonin levels in vivo. Earlier complete lack of retinally derived melatonin or only to
attempts to quantify this indoleamine in skin samples a reduction, remains to be clarified. In other mamma-
by liquid chromatography failed to demonstrate lian taxa, such as rodents, and in non-mammalian
melatonin, and HIOMT activity remained below vertebrates, HIOMT is sufficiently expressed in the
detection levels [35], contrary to cultured HaCaT retina, and melatonin exhibits a nocturnally peaking
keratinocytes, which contained very high amounts (ca circadian rhythm. However, the rate limitation by
2004 R. Hardeland Melatonin, hormone and more

AA-NAT was also disputed for the nocturnal rise of of this process can explain a rapid enzyme inactivation
melatonin in the rat pineal [44]. Because of nocturnal observed upon exposure to nocturnal light, a major
NAS concentrations exceeding those of melatonin component of the photic turnoff mechanism, which
and findings in a low-activity AA-NAT mutant may, however, be accompanied by a shutoff at the
(H28Y), these investigators concluded that HIOMT level of gene transcription.
becomes rate-limiting at night. The two mechanisms of regulation, which act on either
The premier significance of AA-NAT – recently called gene expression or enzyme stability, are effective to a
the timezyme [7] – as the rate-limiting enzyme of different extent among species, and additional varia-
melatonin biosynthesis, e.g., in pinealocytes, deserves tions become apparent between melatonin-synthesiz-
an outline of its regulation. AA-NAT can be regulated ing organs. The AA-NAT phosphorylation/14-3-3
at different levels, that of gene expression and that of mechanism is present in all vertebrates, at least in
enzyme activation and stability (Fig. 1). Which of the pineal gland, whereas this cannot be the case in
these mechanisms is responsible in a specific case, or non-vertebrate organisms which lack the N- and C-
even both of them, depends on species and organ. Two terminal flanking regions [7]. Other post-translational
different types of control have to be distinguished, modifications and interactions with different proteins
which can complement each other under natural that may shield modified amino acid residues have not
conditions: (i) a circadian regulation, either via been studied outside the vertebrates. Data on tran-
components of the oscillator or by norepinephrine- scriptional control of non-vertebrate AA-NAT ho-
dependent rhythms of protein kinases, and (ii) a photic mologs or paralogs are largely missing (except of
turnoff mechanism. Transcriptional regulation is af- processes related to cuticle formation), and the wide
forded by respective control elements in the AA-NAT distribution of N-acetyltransferases indicates other
promoter, especially photoreceptor conserved ele- functions not related to melatonin, but, e.g., to
ments (PCEs; RCS in Drosophila) [45, 46], which sclerotization. Even within the mammals, fundamen-
respond to transcription factors of the CRX/OTX tal differences exist, so that findings obtained in
family [46, 47], and E-boxes, which can be under direct rodents are not generally applicable to humans. In the
control of an oscillator, e.g. by binding of BMAL1/ pineal gland of primates and ungulates, AA-NAT
CLOCK or, in humans, BMAL1/MOP4 heterodimers seems to be only regulated by formation and decay of
[7, 48]. E-boxes can also be accessible to other the phospho-AA-NAT/14-3-3 complex [7], as far as is
transcription factors or modulated by additional known. This is sufficient for a considerable nocturnal
control elements [48]. In mammalian pineals, calci- rise in active enzyme protein as well as the photic
um/cAMP response elements (CREs) can be involved shutoff taking place as soon as cAMP decreases and,
in sympathetic stimulation [7, 49 – 51]. consequently, PKA activity. In rhesus monkeys, a 4-
Regulation at the level of the translated enzyme fold nocturnal increase in AA-NAT activity was
protein comprises phosphorylation and interaction of observed, whereas its mRNA did not show substantial
the phosphorylated AA-NAT with a 14-3-3 protein variations [7]. Similar findings were made in sheep
(isoforms z or e). Both cAMP and Ca2+ signaling can and cattle [7, 54]. However, rodents exhibit a rhythm
contribute to this mechanism, involving either protein in pineal AA-NAT mRNA of remarkable amplitude,
kinase A (PKA) [7, 51, 52] or protein kinase C (PKC) with 100- or even 150-fold nocturnal rises [7, 49, 55,
[53]. Phosphorylation depends on N- and C-terminal 56].
flanking regions, which are typical for the vertebrate The rhythm of AA-NAT mRNA as well as an
AA-NAT, but not found in homologs or paralogs of additional shutoff mechanism at the expression level
urochordates, invertebrate animals or other organims have turned out to be multifaceted phenomena, which
[7]. These regions contain PKA- and PKC-specific do not seem to be solely explained by the rate of
motifs. The association of AA-NAT with a 14-3-3 transcription, although this is, of course, an important
protein (usually studied with subform z) results in basis. Transcription is, again, subject to both positive
stabilization of the enzyme protein [7, 52]. Scaffolding and negative control. Expression can be regulated
with 14-3-3 protein may also favor the stability of the either directly by a circadian oscillator, via tran-
enzymes active center [7]. The stoichiometry of the scription factors mentioned above, or indirectly via a
complex (one or two AA-NAT molecules with one 14- neuronal pathway which is typical for mammals, but
3-3z dimer) is still uncertain. It is of particular contributes to a certain extent also to the pineal
functional significance that the stability of the com- rhythm in birds. In the directly light-sensitive pineal
plex remains moderate, so that it regularly dissociates glands of non-mammalian species and in retinal
and AA-NAT becomes accessible to protein phos- photoreceptors, an autonomous, primary circadian
phatases [52]. The dephosphorylated enzyme is prone oscillator is present in AA-NAT-expressing cells [7,
to rapid proteasomal degradation [7, 54]. The velocity 57, 58], so that this enzyme can be under direct control
Cell. Mol. Life Sci. Vol. 65, 2008 Review Article 2005

Figure 1. Regulation of AA-

NAT expression and activity.
The scheme combines different
pathways which are not collec-
tively present in any one system.
Black arrows and lettering: path-
ways present in pineal glands of
many mammals; violet color:
pathways present in rodents, but
not (or not demonstrated) in
primates and ungulates; brown
color: pathways present in sys-
tems containing endogenous,
self-sustained oscillators (non-
mammalian systems and some
mammalian retinas). Pathways
of particular importance are em-
phasized by larger letters. AA-
NAT, arylalkylamine N-acetyl-
transferase; ai, as, aq, a subunits
of respective G proteins; a1AdR,
a1-adrenergic receptor; b1AdR,
b1-adrenergic receptor; CaMK,
calcium/calmodulin-dependent
kinase; CRE, calcium/cAMP re-
sponse element; CREB, CRE-
binding protein; Glu, glutamate;
hnRNP, heterogeneous nuclear
ribonucleoprotein; ICER, indu-
cible cAMP early repressor;
IRES, internal ribosome entry
site; mGlu3, metabotropic gluta-
mate receptor type 3; NE, nor-
epinephrine; NPY, neuropeptide
Y; PACAP, pituitary adenylyl
cyclase activating peptide; PCE,
photoreceptor conserved ele-
ment; PKA, protein kinase A;
PKC, protein kinase C; PLCb,
phospholipase Cb; TFs, tran-
scription factors; utr, untranslat-
ed sequence; VIP, vasoactive in-
testinal polypeptide; VPAC1,
VIP/PACAP receptor-1; Y1,
NPY receptor-1.

of the oscillator. In addition to transcription factors innervates the pineal [59]. Norepinephrine released
that are part of the clockwork, the oscillator can also from the post-ganglionic sympathetic fibers acts via
regulate AA-NAT expression by modulating cAMP two signaling pathways, which complement and re-
levels, as shown for avian retinas [57, 58]. inforce each other. Binding to b1-adrenergic receptors
The mammalian pineal gland is controlled by the leads to activation of adenylyl cyclase and, conse-
suprachiasmatic nucleus (SCN), a hypothalamic struc- quently, PKA [7, 51, 55], whereas a concomitant
ture acting as a circadian master clock. The photic action via a1B-adrenergic receptors causes, via Go and
input is mediated via the retinohypothalamic tract, the phospholipase Cb, a rise in Ca2+[i] , membrane trans-
paraventricular nucleus and a sympathetic connection location and activation of PKC [51, 53]. Moreover,
from the intermediolateral cell column of the upper elevated cAMP and Ca2+[i] are thought to open Ca2+
thoracic cord to the superior cervical ganglion, which channels – although pineal L-channels were shown to
2006 R. Hardeland Melatonin, hormone and more

be shut by cAMP or adrenergic stimulation [60] – and maximum/minimum ratio of 7 [56]. This indicates a
to further stimulate cAMP production via activated control of AA-NAT mRNA degradation especially by
CaM kinase [51], which can be additionally upregu- hnRNP L. However, the physiological meaning of the
lated via a1B-adrenergic signaling. Both b1- and a1B- IRES-dependent translational upregulation by
adrenergic effects jointly regulate the phosphoryla- hnRNP Q, which contrasts with the negative effect
tion of AA-NAT and, thus, the stabilization by 14-3- on mRNA stability, requires further elucidation.
3z. In systems with transcriptional control, they also
stimulate AA-NAT expression by phosphorylation of
CREB (Ca2+/cAMP response element-binding pro- Melatoninergic actions via membrane receptors
tein), in particular, via PKA type II [7, 51, 61].
The adrenergic cAMP/Ca2+ regulatory system is Since the pioneering work by S. M. Reppert and
further modulated by several neuropeptides and colleagues, who first cloned a melatonin receptor from
glutamate. Pituitary adenylyl cyclase-activating pep- Xenopus melanocytes [64] and who found homologs
tide (PACAP), originating from a trigeminal innerva- in other vertebrate species [65 – 67], high-affinity
tion, and vasoactive intestinal peptide (VIP), from a membrane receptors have been studied in detail. In
pterygopalatine connection, stimulate adenylyl cy- mammals, two receptors are present, MT1 and MT2 (in
clase via a common VPAC1 receptor [51, 60]. Inhib- earlier terminology called Mel1a and Mel1b) [66 – 68].
itory actions are mediated by neuropeptide Y (NPY) The original terminology is sometimes maintained in
from both sympathetic nerve endings and the inter- studies on non-mammalian vertebrates or in compa-
geniculate leaflet, via Gi-coupled pinealocyte Y1 and rative work, because a third melatonin receptor type is
presynaptic Y2 receptors, and from intrapineal gluta- present in these organisms [69], Mel1c, to which no
mate via the metabotropic receptor mGlu3 [51]. denomination corresponding to MT1 or MT2 is
Transcription of the Aa-nat gene, as induced by allowed, for formal reasons of nomenclature. Func-
phospho-CREB (pCREB), is subject to two kinds of tionally different isoforms of Mel1c have been descri-
inhibition. The first represents a negative feedback bed, which lead to decreases in cyclic GMP [70]. Mel1c
mechanism. pCREB also induces a truncated CREM must not to be confused with a binding site formerly
(CRE modulator) variant, ICER (inducible cAMP called MT3, which is an entirely different protein and
early repressor), which interferes with pCREB bind- not a receptor in the strict sense.
ing to its response element [51, 55, 61]. Through this As far as can be judged to date, all chronobiotic
mechanism, Aa-nat expression becomes temporally effects, i.e. all actions directly influencing a circadian
limited. The other type of inhibition seems to be master clock, are mediated by the membrane recep-
specific for the photophase when pineal melatonin has tors mentioned, in mammals by MT1 and MT2 [67 –
to be suppressed. The transcription factors DREAM 69]. In the SCN, melatonin affects, under both in vivo
[downstream regulatory element (DRE) antagonist and in vitro conditions, the phase as well as the
modulator] and the related KChIP (potassium chan- amplitude of the circadian oscillation. Phase shifting is
nel-interacting protein) are capable of downregulat- preferentially exerted via MT2, whereas neuronal
ing the expression of genes containing DRE in their firing is acutely suppressed through MT1 [67, 71, 72].
promoter, in particular Aa-nat, but also Icer [51, 62]. The two receptor subtypes are complementary in their
AA-NAT expression is not only controlled at the actions and can, to a limited extent, mutually sub-
transcriptional level, but in addition post-transcrip- stitute for each other. This seems important insofar as,
tionally by heterogeneous nuclear ribonucleoproteins, exceptionally, a functionally active MT2 (Mel1b) re-
hnRNPs R, Q and L. The effects of these proteins are ceptor can be missing, as found in Siberian hamsters,
dual, depending on their binding sites. When binding Phodopus sungorus and P. campbelli [73]. Since the
to loops in the 3’-untranslated region (3’UTR) region mammalian pineal gland is under control of the SCN,
of the rat AA-NAT mRNA, all three of them strongly the action of melatonin on this circadian pacemaker
favor mRNA degradation [56]. However binding of represents a feedback mechanism involved in the
hnRNP Q to the IRES motif (internal ribosome entry readjustment of the oscillator.
site) in the 5’UTR region stimulates the rate of In seasonal breeders, high densities of the membrane-
translation [63]. In rat pineals, all three hnRNPs bound receptors are also found in the median
exhibited robust circadian rhythms [56]. Levels of eminence and pars tuberalis. While many earlier
hnRNP R and Q were shown to peak (maximum/ reports deal with binding of 2-[125I]iodomelatonin,
minimum ratio ca 2) around the maximum of pCREB specific MT1 expression was demonstrated in various
[zeitgeber time (ZT) 19], about 2 h after the maximum species, whereas the evidence for the presence of MT2
of AA-NAT mRNA (ZT 17), whereas hnRNP L is mostly indirect, based on immortalized cell lines or
expression was maximal 4 h later (ZT 21), with a antagonist experiments [68, 74 – 76]. Melatonin re-
Cell. Mol. Life Sci. Vol. 65, 2008 Review Article 2007

ceptors in the median eminence/pituitary system play Go [79]. PLCb activation has been observed in SCN
a pivotal role in seasonal control, especially of sexual slices, too [83], but this seems to be a more general
activity [65, 68, 75, 76]. This includes an influence of phenomenon of either parallel or alternate melatonin
daytime information by regulation of clock genes in signaling, observed in various target tissues, involving
the pars tuberalis [75]. While melatonin regulates the either pertussis toxin-sensitive (Gi/Go) or insensitive
secretion of hypothalamic and adenohypophyseal (e.g., Gq) G proteins [79, 84, 85]. Actions via PLCb can
hormones, melatonin receptor expression can also have numerous consequences, from activation of PKC
be subject to control by gonadotrophin-releasing subforms, CaM kinases, the opening of Ca2+-activated
hormone, at least in a developmental context. De- K+ channels and modulation of various other protein
pending on vertebrate taxa or species, the effects of kinases of the MAP and JNK pathways. However, it
melatonin on reproductive functions are, sometimes, should be emphasized (i) that the signaling mecha-
much more complex and involve actions at various nisms are strongly cell type-dependent because of
levels of the hormonal axis, including the gonads [68]. differences in G protein subforms and in coupling of
While MT1 and MT2, or – in birds – Mel1c, are highly MT1 or MT2, (ii) that findings obtained in the SCN are
expressed in the SCN or in the median eminence/ not always applicable to peripheral organs and vice
anterior pituitary system, they are also found – versa, and (iii) that the signaling via MT1 or MT2 can
sometimes in a species-specific manner – in various even have opposite effects. An impressive example is
other tissues, where they can exert numerous effects, vasomotor control by melatonin. While actions via
which may be chronobiological in nature, owing to the MT1 cause a pertussis toxin-sensitive vasoconstriction
cyclicity of circulating melatonin, but not necessarily by opening of BKCa channels, those via MT2 result in
chronobiotic, to the extent that master clocks are not vasodilation [20, 68].
involved. The receptors have been detected in many Several melatoninergic agonists and antagonists have
tissues, such as retina, other brain areas, choroid been developed and used as investigative drugs or, in
plexus, cerebral and peripheral vasculature, Harder- the case of ramelteon, as an FDA-approved sleeping
ian gland, reproductive organs including myometrium pill. For reasons of space, this important aspect cannot
and adrenal cortex [68, 76]. Although expression be discussed here in detail.
levels remained low in some of these tissues, their
ample distribution indicates melatonin responsive-
ness of numerous peripheral organs [76]. Other binding sites
The – demonstrated or assumed – functional signifi-
cance of melatonin signaling via membrane receptors In addition to the G protein-coupled receptors, other
cannot be outlined here in any detail for all these binding sites of melatonin exist. Their binding affin-
organs. Instead, with a few selected cases in focus, ities are mostly lower than those of MT1 and MT2, but
signal transduction pathways will be discussed. MT1, could suffice for physiological responses at elevated
MT2 and likewise Mel1c, are G protein-coupled concentrations that seem to be present in several
receptors [64 – 66, 68]. Their classic mode of action is tissues. In at least one case, this extends beyond the
that of a Gi-mediated inhibition of adenylyl cyclase, animals, so that high concentrations of melatonin
resulting in decreases of PKA activity and CREB found in unicells, fungi and some plants may act via
phosphorylation. This holds undoubtedly for SCN non-membrane binding sites.
neurons, but is, even there, not an exclusive mecha- One of the melatonin-binding proteins was originally
nism. As with many other G protein-coupled recep- believed to be another membrane receptor and named
tors, the presence of G protein subforms in a particular MT3, but it turned out to be a mainly cytosolic enzyme,
cell type can substantially change the specific re- quinone reductase 2 (= QR2 = NRH:quinone oxido-
sponse. Co-activation or alternate activation of Go or ACHTUNGREreductase 2 = NQO2; NRH = dihydronicotinamide
Gq has been repeatedly observed [77 – 79]. In some riboside) [86 – 88]. The enzyme is expressed in several
cases, including studies in transfected cells, other G tissues, including the brain [87]. Some of its polymor-
proteins such as Gz or G16 were reported to couple phic subforms have been related to Parkinsons
differentially to melatonin receptors [79 – 81]. While disease [89]. Disruption of the NQO2 gene leads to
coupling to Gi protein subforms causes decreases in bone marrow myeloid hyperplasia [90]. This indicates
cAMP, rises in cAMP were also described, e.g., for a role beyond detoxification of xenobiotics. The
Mel1c signaling via az coupling to adenylyl cyclase type frequently proposed assumption of a role in redox
II [82]. Gi-dependent mechanisms may affect not only metabolism, eventually in terms of protection, is
cAMP levels, but may also modulate, in some cells, K+ actually nothing more than an idea. Even though a
conductance, and additionally stimulate, via bg, phos- function in ubiquinone reduction has been suggested,
pholipase Cb (PLCb), which may also be the case with the precise role of this enzyme is not really understood
2008 R. Hardeland Melatonin, hormone and more

[91]. Recently, melatonin was assumed to be a co- Although a considerable amount of clarification is
substrate serving as a hydrogen/electron donor to required concerning the binding sites different from
other redox co-factors such as FAD [92], a concept the G protein-coupled receptors, the findings men-
still requiring further experimental support. It should tioned in this section collectively indicate that mela-
also be noted that NAS has an affinity for NQO2 tonin is much more pleiotropic than previously
comparable to that of melatonin [cf. discussion in ref. believed, not only with regard to the numerous target
20]. organs, G proteins and G protein-regulated proteins
A melatonin-binding protein of considerable regula- involved, but also to additional signaling mechanisms.
tory significance is calmodulin. Its affinity to melato-
nin is sufficient for mediating effects at elevated
physiological concentrations, especially those at- Tissue versus circulating melatonin
tained in tissues [93 – 95]. Melatonin binding results
in inhibitions of CaM kinase II [94] and of neuronal The consequences of extrapineal melatonin biosyn-
NO synthase [96]. Moreover, melatonin causes thesis merit particular consideration because they
PKCa-dependent phosphorylation of calmodulin profoundly change our view of the biological role of
[97], presumably by signaling mechanisms described this molecule. Absence of robust melatonin rhythms
in the preceding section, but this effect is important or low-amplitude variations imply roles different from
insofar as it perpetuates CaM-dependent inhibitions. the transmission of dark signals. In tissues containing
Interacting calmodulin and kinase effects are relevant considerably higher levels than in the circulation, a
to rearrangements of the cytoskeleton [95], which protective role may be assumed. Numerous antiox-
represent some of the earliest effects described for idant, antiinflammatory, antiexcitatory/antiexcitotox-
melatonin, including ciliates and plants [8, 96]. An ic and oncostatic effects have been reported in
additional facette of melatonin/Ca2+ interactions is countless publications and have been frequently
the binding to calreticulin, and, perhaps, to two reviewed [8, 20, 27, 31, 91, 96, 102, 104, 105]. Although
nuclear proteins, one of which had high homology to many of these investigations have been conducted at
calreticulin, whereas the other was structurally differ- high doses, they are not collectively physiologically
ent [98]. irrelevant, especially when tissue concentrations are
Nuclear binding sites of melatonin have been a matter approaching the micromolar range, or when protec-
of considerable debate. Meanwhile, many publica- tion by melatonin has been demonstrated at levels
tions have dealt with transcription factors belonging to found in the circulation [20, 96].
the retinoic acid receptor superfamily, in particular, In organs which contain melatonin above plasma
RORa1, RORa2 and RZRb [99 – 102], and their levels, but poorly release the indoleamine, an answer
classification as nuclear receptors seems justified, has to be given as to why melatonin is not – or only in
although their affinity to melatonin is lower. A low quantities – entering the circulation, although it is
synthetic ligand, CGP 52608 [99], has been repeatedly usually believed to cross any membrane because of its
used for identifying effects by these nuclear proteins. amphiphilicity [27, 91, 105]. If tissue concentrations
RORa1 and RORa2 seem to be involved in some have been determined correctly, the answer can only
aspects of immune modulation, and RZRb is ex- be that of sequestration by non-receptor binding sites
pressed in the central nervous system, including the [20]. It cannot be said with certainty whether the
pineal gland [20, 102]. Moreover, RORa was assumed major proteins involved have already been identified,
to mediate upregulations of antioxidant enzymes but the binding to calreticulin [98] and to abundant
[101]. Still, the full spectrum and physiological mean- nuclear proteins [106] may contribute to melatonin
ing of these receptors remains to be clarified. retention.
A further melatonin binding site seems to exist in rat Low-amplitude rhythms or absence of (robust)
brain mitochondria, for which a dissociation constant rhythms in some tissues can resolve another paradox,
of 150 pM and a total number of specific binding sites namely, why nocturnally peaking melatonin should
of 30 fmol/mg have been determined [20]. The protein exert protective effects in both day- and night-active
is assumed to be localized at the amphipathic ramp of animals, whereas reactive oxygen and nitrogen species
complex I in the mitochondrial electron transport are preferentially produced in circadian phases of
chain. At elevated concentrations, melatonin was also motor and neuronal activity. If rhythmicity is poorly
shown to directly inhibit the opening of the mitochon- expressed in an organ or almost absent, the circadian
drial permeability transition pore [103], a finding that phase loses its relevance for tissue melatonin, and only
would imply an additional, low-affinity mitochondrial the generation of reactive intermediates becomes
binding site. decisive for a phase of detoxification [91, 105].
Cell. Mol. Life Sci. Vol. 65, 2008 Review Article 2009

High amounts of tissue melatonin – cf. the several butions, CYP1A1 and CYP1B1 [110], are present in
hundred-fold quantities in the gastrointestinal tract the liver, but are found in other tissues too. The major
compared to the pineal – has another important demethylating subform, CYP2C19, is also expressed
consequence, which is at variance with the frequently in the liver [110]. After oral melatonin administration
read statement that melatonin is almost quantitatively to mice, NAS amounted to only about 3 % of the
metabolized in the liver to 6-hydroxymelatonin, with urinary metabolites [113].
subsequent conjugation and excretion as 6-sulfatox- Deacetylation to 5-MT leads to another bioactive
ymelatonin. This may be true for the circulating molecule. Contrary to melatonin and NAS, this
hormone and for single intravenous injections, but has molecule is a substrate of monoamine oxidase A
already turned out to be wrong in the infusion (MAO A), and the resulting oxidation product, 5-
experiment [28]. Release of non-metabolized mela- methoxyindole-3-acetaldehyde, is converted by alde-
tonin to the intestinal lumen has been shown repeat- hyde dehydrogenase to 5-MIAA, or by alcohol
edly [26, 28 – 30]. With regard to the manyfold higher dehydrogenase to 5-ML (Fig. 2), two compounds
quantities outside pineal and circulation, hepatic 6- which are also formed by O-methylation of 5-
hydroxylation as an almost exclusive pathway appears hydroxylated analogs [20]. These metabolites are
extremely illogical. That statement also negates long- not restricted to vertebrates, but are also produced
known alternate pathways of melatonin metabolism in dinoflagellates and, 5-ML at least, in brown algae,
(Fig. 2), such as demethylation by the P450 isoforms red algae and yeast [8, 10, 42]. While 5-MIAA seems
CYP2C19 and, to a much smaller extent, CYP1A2; to be an end product, which is excreted, 5-ML has
deacetylation by aryl acylamidases, melatonin deace- been considered as another bioactive molecule. A
tylases or eserine-sensitive acetylcholinesterase, and further 5-MT metabolite, the b-carboline pinoline,
pyrrole-ring cleavage [20, 102]. Their relevance in the known as a psychotropic drug interfering with sero-
central nervous system was recently reviewed [20]. In tonin availability, will not be discussed here because a
the spinal cord, an inhibitory effect of melatonin on considerable fraction is artificially formed during
nociceptive transmission was maintained by eserine extraction.
[107]. In Xenopus melanophores, a loss of responsive- N,N-dimethyl-5-methoxytryptamine, which is either
ness to melatonin was prevented by the same drug produced from 5-MT or by O-methylation of bufote-
[108]. Other recent estimations of quantitative mela- nin, has properties of an endogenous hallucinogen
tonin metabolism came to the conclusion that about [20]. O-acetyl-5-methoxytryptophol, a structural mel-
one-third of the indoleamine is converted by pyrrole- atonin homolog, is formed from 5-ML in organs
ring cleavage to methoxylated kynuramines (Fig. 2) containing high melatonin concentrations, such as the
[109]. The rates may be even higher in some tissues in pineal gland. Several pharmacological effects com-
which P450 enzymes are poorly expressed [27]. These prise inhibition of nicotinic and muscarinic acetylcho-
kynuramines had already been shown decades ago to line receptors as well as decreases in pituitary
represent major melatonin metabolites in the brain, prolactin and luteinizing hormone [20].
whereas no 6-hydroxymelatonin was detected in that The metabolic route of pyrrole-ring cleavage (Fig. 2)
study [22], although 6-hydroxylation should be possi- leads to the 5-methoxylated kynuramines, N1-acetyl-
ble in the brain because of the presence of CYP1B1 N2-formyl-5-methoxykynuramine (AFMK) and N1-
[110] and may have been overlooked at that time. acetyl-5-methoxykynuramine (AMK). AFMK can be
These findings are of particular importance insofar as formed either directly from melatonin or indirectly
melatonin is released from the pineal gland not only to from a tricyclic metabolite, cyclic 3-hydroxymelatonin
the circulation, but also, at much higher concentra- (c3OHM). c3OHM is a product of melatonin oxida-
tions, via the pineal recess into the third ventricle [111, tion by free radicals, in particular, by sequential
112]. Since melatonin concentrations in the cerebro- interactions with two hydroxyl radicals [114, 115]. It
spinal fluid obtained from lumbal puncture are in the is found in rodent urine after melatonin administra-
range of serum levels, most of the compound must tion [113], and is strongly elevated after exposure to
have been either taken up or metabolized by the brain ionizing radiation [114]. It is a remarkable fact that
tissue before entering the spinal cord. numerous reactions of melatonin lead to the same
product, AFMK, and that this kynuramine is fre-
quently the major product in various oxidation
Complexity of metabolism systems, especially when they are designed in a way
not to generate a single radical species, but to consider
P450 enzymes are organ specifically expressed and the physiological prevalence of superoxide anions
catalyze hydroxylation or dealkylation. The 6-hydrox- [115, 116], which can serve as terminators of radical
ylating subforms, CYP1A2, and with smaller contri- reaction chains [8, 115, 116]. The spectrum of reac-
2010 R. Hardeland Melatonin, hormone and more

Figure 2. The complexity of mel-

atonin metabolism. AAA, aryl
acylamidase; AA-NAT, arylal-
kylamine N-acetyltransferase;
AChE, acetylcholinesterase;
ADH, alcohol dehydrogenase;
AldDH, aldehyde dehydrogen-
ase; CYP, cytochrome P450 iso-
form (monoxygenase or dealky-
lase); HNO, nitroxyl; MelDA,
melatonin deacetylase; NATs,
N-acetyltransferases (other than
AA-NAT); ·NO, nitric oxide rad-
ical; OAT, O-acetyltransferase;
·OH, hydroxyl radical; ROS, re-
active oxygen species. *Full
name: N-(1-formyl-2-hydroxy-5-
methoxy-3-oxo-2,3-dihydro-1H-
indol-2-ylmethyl)-acetamide; ad-
ditional non-hydroxylated and
deformylated 3-indolinones
have been identified as AFMK
metabolites and chemically char-
acterized [123]. Additional reac-
tions and metabolites, which also
exist, have not been included.

tions leading to AFMK is exceptional. It comprises experiments or in untreated animals should be seen in
enzymes, such as indoleamine 2,3-dioxygenase or, relation to administration routes, formation in tissues
quantitatively important, myeloperoxidase [109], and the metabolic fate of the kynuramines, which may
pseudoenzymatic catalysis by hypervalent oxyferryl- not become apparent in body fluids. High concen-
hemoglobin or hemin, and various photochemical and trations of AFMK have been found in HaCaT
radical reactions [10, 105, 117]. keratinocytes [36]. One might also note high amounts
The quantitative relevance of AFMK and AMK and circadian rhythmicity of AFMK found in a plant,
seems to have been underrated for quite some time. the water hyacinth, Eichhornia crassipes [118]. At
Apart from the widely neglected demonstration of lower physiological concentrations of melatonin, the
these kynuramines as major melatonin metabolites in role of its oxidation by free radicals may largely be
the brain [22], it may be a misconception to judge their seen in the formation of AFMK and its secondary
significance on the basis of urinary or plasma concen- metabolites [91, 105].
trations. Only small amounts of AFMK and AMK Other products deriving from melatonin by radical
were found in the urine upon oral administration of reactions are hydroxylated indoles, whereas dimers
melatonin [113], but, in stark contrast, high quantities are relatively rare because of the methoxy group,
were found after injection into the cisterna magna which largely prevents the formation of O-centered
[22]. The low urinary quantities detected in some and C-centered indolyl radicals [22, 105]. Preferential
Cell. Mol. Life Sci. Vol. 65, 2008 Review Article 2011

sites of hydroxylations have been identified [119]. of hydroxyl radicals by this indoleamine [128]. How-
Apart from c3OHM, 6-hydroxy- and 2-hydroxymela- ever, antioxidative protection has turned out to be
tonin can be formed, the latter being in equilibrium more than radical scavenging. Under physiological
with its tautomer, N-acetyl-5-methoxy-2-indolinone conditions, the capability of donating electrons to free
[120]. radicals – which is present in this molecule without any
Deformylation of AFMK to AMK is catalyzed by two doubt – may have other biological meanings than to
enzymes, arylamine formamidase [27, 102, 105] and detoxify a certain number of free radicals, already
hemoperoxidase (catalase) [105, 121]. Recently, from the viewpoint of stoichiometry. At pharmaco-
another photochemical mechanism by UV light logical concentrations, the value of melatonin as a
(Fig. 2) has been described [122]. AMK formation direct scavenger has been repeatedly demonstrated
may not be an exclusive route of AFMK metabolism, (cf. reviews mentioned). Certain cells or organisms
since free-radical reactions also led to a couple of C2- containing melatonin concentrations in the (some-
substituted 3-indolinones (Fig. 2), representing a times upper) micromolar range may also profit by
novel class of oxidation products [123]. direct scavenging. Additional roles may be sought in
While AMK was regarded for decades as an end the non-enzymatic formation of other bioactive com-
product because of its appearance in the urine, this pounds and in the capability of undergoing reactions
seems rather unlikely, since AMK was shown to be with electron exchanging or transporting systems,
easily oxidized [124] and to readily interact with such as the respiratory chain [91, 105]. Upregulation
reactive nitrogen species. Even the dry solid forms of antioxidant and downregulation of – a few –
new products with trace gases present in the air, when prooxidant enzymes [91, 104, 105] certainly contrib-
exposed on the large surface of a silica gel. Two utes to the antioxidant balance, but these effects
products are formed in solution by reactive inter- should not be generally overrated. Induction of
mediates present in biological material, a nitrosated hemoperoxidase (catalase), superoxide dismutase
derivative, 3-acetamidomethyl-6-methoxycinnolinone subforms, glucose-6-phosphate dehydrogenase and
(AMMC) (Fig. 2), and a nitrated compound, N1- g-glutamylcysteine synthase are organ-specific, and
acetyl-5-methoxy-3-nitrokynuramine (AMNK = 3- the meaning of upregulations of just a few percent,
nitro-AMK) [125]. AMMC is generated by all three sometimes only demonstrated at the mRNA level, is
different NO congeners, ·NO, NO+ and HNO, al- questionable. A most frequently reported and reliable
though the reaction with NO+ is physiologically less effect is the stimulation of glutathione peroxidase, as
likely, because of the short half-life of the cation in particularly found in the central nervous system. The
aqueous solution at pH 7.4 [126]. AMMC represents a cellular mechanism of upregulation has only been
stable compound, contrary to the majority of other N- tentatively addressed [101], but is not really elucidat-
nitrosated substances [127], including N-nitrosomela- ed in its details.
tonin, which easily redonate NO. AMNK was found to Since detoxification of free radicals and other oxi-
be produced by the combination of peroxynitrite and dants did not fully explain the antioxidative efficacy of
CO2, a physiological nitrating mixture leading to melatonin, radical avoidance was brought into focus
carbonate radicals (CO3·) and ·NO2 [125]. [27, 91, 105]. Formation of reactive oxygen and
nitrogen species can be reduced by several actions of
melatonin. First, appropriate timing and coordination
The non-classic actions of melatonin of rhythms, as favored by the indoleamine, should
diminish oxidative stress, because oxidative damage
Countless publications have dealt with protective was elevated in the clock mutants, per0 and pers in
actions of melatonin. Although the number of such Drosophila, tau in Syrian hamster [91]. Second, the
reports is meanwhile higher than that on its chrono- antiexcitatory/antiexcitotoxic effects of melatonin
biological role, the understanding in mechanistic should reduce radical formation as well, by attenuat-
terms still awaits further deepening under many ing their metabolism-related generation and the
aspects. Nevertheless, the high potential of such a prevention of Ca2+- and NO-dependent cellular stress
function seems worthy of considerable efforts. [91, 105]. In this context, mitochondria seem to be a
Melatonin participates in various lines of defense. A particular target. These organelles represent in many
first one is that of antioxidative protection, a phenom- cells a major source of superoxide anions, due to
enon which has been frequently reviewed [e.g., 8, 27, electron leakage, especially from complexes I and III
91, 96, 101, 102, 104, 105, 119, 121] and will be of the electron transport chain, radicals which are not
discussed here only in its general traits. Investigation quantitatively eliminated by mitochondrial and cyto-
of the antioxidant actions of melatonin was strongly solic superoxide dismutases (MnSOD and Cu,Zn-
stimulated by the finding of potent direct scavenging SOD). Superoxide anions are sources of peroxynitrite
2012 R. Hardeland Melatonin, hormone and more

from cytosolic or mitochondrially generated NO and natural killer (NK) cells and several leukocyte-de-
lead, via combination with CO2 or protons, to rived cell lines [37, 40, 140]. It was also found in
carbonate (CO3·), hydroxyl (·OH) and ·NO2 radicals. thymocytes and epithelial cells [40], but the origin is
H2O2 formed by the SODs is a source of hydroxyl not clear in these cases. Immunomodulation by
radicals. Electron overflow at the iron-sulfur cluster melatonin has recently been reviewed several times
N2 of complex I, i.e., at the bottleneck of the transport [e.g., 40, 102, 131, 132]. Main findings are the
chain, seems to be an important cause of enhanced activation of various cell types, such as T, B and NK
superoxide generation. This site is assumed to be cells, monocytes and splenocytes, and modulation of
modulated by melatonin [27, 91, 105] and may be cytokine release. Melatonin was shown to enhance the
associated with a mitochondrial high-affinity binding production of interleukin (IL)-2, IL-6 and IL-12,
site [20] (cf. section on Other binding sites). Although whereas levels of interferon-8 or tumor necrosis-
many details of this concept of mitochondrial radical factor-a were sometimes decreased but in other cases
avoidance remain to be elucidated, pertinent effects of increased [40, 143]. Melatonin also counteracted
melatonin at the level of this organelle have been inhibitory effects of PGE2 on IL-2 production [68,
repeatedly described, such as support of mitochon- 144]. With regard to the complexity of the immune
drial electron flux, stimulation of complex I and IV system, some divergence in effects depending on cell
activities, prevention of mitochondrial calcium over- types, differentiation state and mixtures of cells in test
load and maintenance of mitochondrial membrane systems should not be surprising. In the immune
potential and of ATP formation [91, 105, 129, 130]. system, different melatonin receptors are involved.
These findings gain particular significance because of MT1 was found to mediate effects concerning IL-2, but
normalizations in mitochondrial functions achieved the presence of this receptor was demonstrated in
by melatonin in senescence-accelerated mice [131, numerous leukocyte subtypes [40, 68]. Signaling via
132]. MT2 was, e.g. shown to stimulate splenocyte prolifer-
Mitochondrial hypoactivity and dysfunction are phe- ation and to decrease leukocyte rolling [68, 145]. In
nomena associated with aging, but also with numerous avian splenocytes, growth stimulation is mediated by
diseases. These organelles also play a pivotal role in the Mel1c receptor [146]. Based on pharmacological
the induction of apoptosis. Melatonin has been criteria, inhibition of leukotriene B4-induced endo-
frequently shown to antagonize or prevent apoptosis thelial leukocyte adhesion was ascribed to the binding
by modulating mitochondrial functions [133, 134], site previously named MT3 [145], now known to be
including effects on calcium homeostasis and mito- quinone reductase 2, but, in the absence of demon-
chondrial membrane potential [105, 134, 135], and strated signaling pathways, this should be judged with
also by directly inhibiting the mitochondrial perme- caution. Expression of ROR and RZR subforms has
ability transition pore [103]. been reported for various leukocytes and related cells
Antiexcitatory and antiexcitotoxic effects are partially such as splenocytes, thymocytes and Jurkat cells, and
related to mitochondria, in terms of avoidance of some of the actions of melatonin seem to be mediated
calcium overload and elevated radical formation in by these transcription factors [100, 141, 142, 147].
these organelles. These actions, which extend to High densities of nuclear binding sites, as sometimes
anticonvulsant and anxiolytic properties [102], go observed, should not be immediately taken as a sign
beyond the chronobiotic and sleep-inducing actions for involvement of ROR/RZR receptors, because of
and, again, are highly complex. In mammals, modu- other nuclear binding proteins [98, 106]. In melatonin-
lations of GABA and glutamate signaling are involved synthesizing leukocytes, concentrations may be suffi-
and include secondary effects through decreases in cient for nuclear receptors and mediate autocrine
cytosolic Ca2+ via GABAc [136] or metabotropic effects. The interplay of membrane and nuclear
mGlu3 receptors [137], interference with neuronal NO receptors in the immune system remains an intriguing
synthase, directly, or indirectly by AMK [102, 104 field.
105], effects on K+ currents, as studied in the
cerebellum [138], and potentiation of strychnine-
sensitive glycine-induced currents [139]. Actions of metabolites
Immunomodulation by melatonin represents another
line of defense. This includes antiiflammatory proper- Melatonin may also be regarded as a prodrug leading
ties of the indoleamine, in which antioxidant, NO- to other bioactive molecules [27]. Earlier studies were
attenuating, and mitochondrial effects act in concert mostly oriented at actions already known from
with the attenuation of proinflammatory signaling. melatonin, with the frequent outcome that the metab-
Moreover, melatonin is produced by various leuko- olites were only effective at higher concentrations
cytes, such as monocytes, eosinophils, mast cells, [20]. Substances like 5-MT and NAS, being both
Cell. Mol. Life Sci. Vol. 65, 2008 Review Article 2013

metabolites and precursors, possess high affinities for studied. In cases of viral meningitis, considerably
some 5-HT receptor subforms found in both the enhanced levels of AFMK in the cerebrospinal fluid
central nervous system and the periphery [reviewed in correlated with inflammatory markers such as tumor
ref. 20]. It is, however, difficult to judge a putative role necrosis factor-a, IL-8 and IL-1b [153]. This may only
of these compounds as secondary mediators of reflect rises in cerebral melatonin oxidation. How-
melatonin in the CNS, in part because of concentra- ever, AFMK was also reported to be a more efficient
tions, especially in the case of the easily metabolized 5- inhibitor of lipopolysaccharide (LPS)-induced pro-
MT, and because NAS is apparently formed without duction of TNF-a and IL-8 in neutrophils, compared
being converted to melatonin and exerts independent to melatonin [154], an effect which cannot be ex-
actions. The actions of 5-MT in brain and retina have plained by affinity or selectivity to free radicals
been recently reviewed [20]. More detailed informa- released in response to LPS.
tion on profound and ecophysiologically relevant AFMK was also used to antagonize oxidative stress, at
actions by 5-MT in dinoflagellates can be found pharmacological concentrations. It was shown to
elsewhere [10, 42]. protect DNA from oxidative damage by hydroxyl
Actions of non-indolic metabolites have only been radicals generated by a chromium(III)-based Fenton-
decribed for mammals, except for some unpublished, analog reaction [155], or by a d-aminolevulinic acid/
preliminary data on life extension by AFMK in a Fe2+ system [156], but it remained less efficient than
rotifer, Philodina acuticornis [B. Poeggeler, personal melatonin. This is not surprising because AFMK
communication]. In the first decades after discovery exhibits a preference for two-electron transfer reac-
of AFMK and AMK, some effects were described tions [117] and is, therefore, a poorer radical scavenger
concerning prolactin release, retardation of testicular than melatonin or its deformylated metabolite, AMK,
growth, binding to benzodiazepine and melatonin which easily undergoes single-electron transfer reac-
receptors [summarized in ref. 148]. Binding affinities tions [105, 124]. Thus, protective effects by AFMK
were not sufficient to explain actions of the indole- observed in living cells, such as inhibition of toxicity by
amine by conversion to AFMK or AMK. However, an glutamate, H2O2, or amyloid b25 – 35 peptide in hippo-
interesting chronobiological effect of AFMK was campal neurons [117], might be the consequence of its
described, which was, unfortunately, never followed conversion to AMK. The same may hold for protec-
up in other comparable systems. In rats, AFMK was tion against oxidative damage to DNA, proteins and
shown to promote the re-entrainment of the melato- lipids by X-rays [157]. Protective actions by AMK are
nin rhythm [149]. In the taxonomically very distant more than just radical scavenging, because of addi-
Lingulodinium, no phase shifting by AFMK was tional effects already found at very low, nanomolar or
observed [R. Hardeland, unpublished data]. How- even lower concentrations. Support of mitochondrial
ever, the life cycles of malaria parasites (Plasmodium functions was observed in the nanomolar range [130].
chabaudi and Plasmodium falciparum) were AMK, being an amphiphilic compound with a slightly
synchronized by AFMK in the upper nanomolar higher lipophilicity than melatonin, has been suggest-
range, an effect which was associated with rises in ed to participate in an electron shuttle, which may
cytosolic calcium and which was blocked by luzindole bridge bottlenecks in the electron transport chain and,
[150], otherwise being an MT1/MT2 melatonin recep- thereby, diminish electron leakage [91, 105]. Atten-
tor antagonist. The inhibition by luzindole may raise tion may also be directed to the structural similarities
questions concerning conclusions on melatonin sig- between ubiquinones and AMK [91]. Moreover,
naling via MT1 and/or MT2 in other cases, or on a AMK was shown to be a highly efficient inhibitor of
cryptic, perhaps indirect action of AFMK at these neuronal NO synthase, with an IC50 in the nanomolar
receptors. range, but demonstrable efficacy already at 1011 M
AMK was shown to efficiently inhibit prostaglandin [158].
synthesis [151], a property of, perhaps, considerable With regard to specific actions of kynuramines, it
pharmacological interest. At the time of discovery, should be remembered that this chemical family
cyclooxygenases (COX) 1 and 2 were not distinguish- represents its own, though frequently forgotten, class
ed, but the efficacy of AMK was reported to be far of biogenic amines, including 5-hydroxylated, C5-
higher than that of acetylsalicylic acid (aspirin). unsubstituted, and N,N-dimethylated compounds, for
Recently, AMK was reported to downregulate which various effects have been described [148].
COX-2 – but not COX-1 – expression in macrophages, Other products from the kynuric pathway of melato-
an effect shared by its precursors AFMK and mela- nin metabolism may additionally contribute to a
tonin [152]. Whether antiinflammatory or other complex action spectum. The group of cinnolines, to
immunological actions by 5-methoxylated kynura- which AMMC belongs, contains pharmacologically
mines may be of medicinal relevance, remains to be active compounds, which have been used as medica-
2014 R. Hardeland Melatonin, hormone and more

ments or investigative drugs, because of their anti- 3 Quay, W. B. (1964) Circadian and estrous rhythms in pineal
melatonin and 5-hydroxy indole-3-acetic acid. Proc. Soc. Exp.
allergic, antitumor, anxiolytic and other neurotropic Biol. Med. 115, 710 – 713.
properties [125]. 4 Axelrod, J., Wurtman, R. J. and Winget, C. M. (1964)
Melatonin synthesis in the hen pineal and its control by
light. Nature 201, 1134.
5 Lewy, A. J., Wehr, T. A., Goodwin, F. K., Newsome, D. A. and
Markey, S. P. (1980) Light suppresses melatonin secretion in
Conclusions humans. Science 210, 1267 – 1269.
6 Binkley, S. (1993) Structures and molecules involved in
generation and regulation of biological rhythms in vertebrates
The diversity of melatonin actions, the number of and invertebrates. Experientia 49, 648 – 653.
demonstrated or putative binding sites, differential G 7 Klein, D. C. (2007) Arylalkylamine N-acetyltransferase: the
protein coupling, the existence of numerous target timezyme. J. Biol. Chem. 282, 4233 – 4237.
8 Hardeland, R. and Fuhrberg, B. (1996) Ubiquitous melatonin
cells and organs, pineal and extrapineal formation and – presence and effects in unicells, plants and animals. Trends
the possibility of additional actions by its metabolites Comp. Biochem. Physiol. 2, 25 – 45.
demonstrate an exceptional pleiotropy. This may 9 Hardeland, R. and Poeggeler, B. (2003) Non-vertebrate
reflect an orchestrating function, but may also become melatonin. J. Pineal Res. 34, 233 – 241.
10 Hardeland, R., Pandi-Perumal S. R. and Poeggeler, B. (2007)
a problem when specific actions are desired, in terms Melatonin in plants – focus on a vertebrate night hormone
of both experimental and applied approaches. In with cytoprotective properties. Funct. Plant Sci. Biotech-
practice, multiple effects can hardly be avoided. nol. 1, 32 – 45.
11 Dubocovich, M. L. (1983) Melatonin is a potent modulator of
Whether they are really disadvantagous for a treated dopamine release in the retina. Nature 306, 782 – 784.
organism, or dangerous, remains to be studied. 12 Cahill, G. M. and Besharse, J. C. (1989) Retinal melatonin is
As a substance found in numerous phylogenetically metabolized within the eye of Xenopus laevis. Proc. Natl.
Acad. Sci. USA 86, 1098 – 1102.
very distant organisms, at levels differing considerably 13 Kazula, A., Nowak, J. Z. and Iuvone, P. M. (1993) Regulation
from taxa to taxa, a uniform spectrum of actions of melatonin and dopamine biosynthesis in chick retina: the
should not be expected, although some themes, such role of GABA. Vis. Neurosci. 10, 621 – 629.
14 Buzzell, G. R., Pangerl, A., Pangerl, B., Menndez-Pelez,
as calmodulin signaling or antioxidant actions, may go A., Vaughan, M. K., Little, J. C., Hill, S. M., Vaughan, G. M.
across the multitude of species synthesizing this and Reiter, R. J. (1990) Melatonin and porphyrin in the
indoleamine. Various findings indicate that it is not harderian glands of the Syrian hamster: circadian patterns and
generally a compound found only in traces – as in the response to autumnal conditions. Int. J. Biochem. 22, 1465 –
1469.
circulation. Micromolar levels can be present outside 15 Payne, A. P. (1994) The harderian gland: a tercentennial
the vertebrates, and some findings indicate that tissue review. J. Anat. 185, 1 – 49.
melatonin may be sometimes much higher than the 16 Djeridane, Y. and Touitou, Y. (2001) Melatonin synthesis in
the rat Harderian gland: age- and time-related effects. Exp.
blood level. Additional complexity results from differ- Eye Res. 72, 487 – 492.
ent sites of formation in a vertebrate body, which, 17 Tosini, G. and Menaker, M. (1998) Multioscillatory circadian
again, exceeds the classic sites of biosynthesis. The organization in a vertebrate, Iguana iguana. J. Neurosci. 18,
1105 – 1114.
consequences of tissue melatonin and its different fate 18 Zawilska, J. B., Berezinska, M., Rosiak, J., Skene, D. J.,
from that of the circulating hormone deserves further Vivien-Roels, B. and Nowak, J. Z. (2004) Suppression of
attention. Actions of melatonin metabolites other melatonin biosynthesis in the chicken pineal gland by retinally
perceived light – involvement of D1-dopamine receptors. J.
than 6-hydroxy-/6-sulfatoxymelatonin are a promising Pineal Res. 36, 80 – 86.
field awaiting further research, especially with regard 19 Grace, M. S. and Besharse, J. C. (1993) Solubilization and
to the central nervous system. This holds, in particular, biochemical characterization of the melatonin deacetylase
for major brain metabolites like AMK and its from Xenopus laevis retina. J. Neurochem. 60, 990 – 999.
20 Hardeland, R. and Poeggeler, B. (2007) Actions of melatonin,
derivatives, perhaps also for 5-MT. Investigators its structural and functional analogs in the central nervous
should not simply direct their attention to effects system and the significance of metabolism. Cent. Nerv. Syst.
already known for melatonin, but rather seek for Agents Med. Chem. 7, 289 – 303.
21 Iuvone, P. M., Gan, J. and Alonso-Gmez, A. L. (1995) 5-
independent actions. As melatonin is not only a Methoxytryptamine inhibits cyclic AMP accumulation in
hormone, its levels and those of its metabolites should cultured retinal neurons through activation of a pertussis
not only and primarily be determined in body fluids toxin-sensitive site distinct from the 2-[125I]iodomelatonin
binding site. J. Neurochem. 64, 1892 – 1895.
but rather in tissues, and differences in administration 22 Hirata, F., Hayaishi, O., Tokuyama, O. and Senoh, S (1974) In
routes deserve particular consideration. vitro and in vivo formation of two new metabolites of
melatonin. J. Biol. Chem. 249, 1311 – 1313.
23 Menendez-Pelaez, A, Rodriguez, C. and Dominguez, P.
1 Lerner, A. B., Case, J. D., Takahashi, Y., Lee, T. H. and Mori, (1991) 5-aminolevulinate synthase mRNA levels in the
W. (1958) Isolation of melatonin, the pineal gland factor that Harderian gland of Syrian hamsters: correlation with por-
lightens melanocytes. J. Am. Chem. Soc. 80, 2587 – 2592. phyrin concentrations and regulation by androgens and
2 Ruffin, N. E., Reed, B. L. and Finnin, B. C. (1969) The melatonin. Mol. Cell. Endocrinol. 80, 177 – 182.
specificity of melatonin as a melanophore controlling factor in 24 Matsubara, E., Bryant-Thomas, T., Pacheco Quinto, J.,
the pencil fish. Life Sci. 8, 1167 – 1174. Henry, T. L., Poeggeler, B., Herbert, D., Cruz-Sanchez, F.,
Cell. Mol. Life Sci. Vol. 65, 2008 Review Article 2015

Chyan, Y.-J., Smith, M. A., Perry, G., Shoji, M., Abe, K., 43 Callebert, J., Jaunay, J.-M. and Jallon, J.-M. (1991) Control of
Leone, A., Grundke-Ikbal, I., Wilson, G. L., Ghiso, J., Drosophila biorhythms. Adv. Pineal Res. 5, 81 – 84.
Williams, C., Refolo, L. M., Pappolla, M. A., Chain, D. G. 44 Liu, T. and Borjigin, J. (2005) N-acetyltransferase is not the
and Neria, E. (2003) Melatonin increases survival and inhibits rate-limiting enzyme of melatonin synthesis at night. J. Pineal
oxidative and amyloid pathology in a transgenic model of Res. 39, 91 – 96.
Alzheimers disease. J. Neurochem. 85, 1101 – 1108. 45 Kikuchi, T., Raju, K., Breitman M. L. and Shinohara, T.
25 Huether, G. (1993) The contribution of extrapineal sites of (1993) The proximal promoter of the mouse arrestin gene
melatonin synthesis to circulating melatonin levels in higher directs gene expression in photoreceptor cells and contains an
vertebrates. Experientia 49, 665 – 670. evolutionarily conserved retinal factor-binding site. Mol. Cell.
26 Bubenik, G. A. (2002) Gastrointestinal melatonin: localiza- Biol. 13, 4400 – 4408.
tion, function, and clinical relevance. Dig. Dis. Sci. 47, 2336 – 46 Appelbaum, L., Vallone, D., Anzulovich, A., Ziv, L., Tom,
2348. M., Foulkes, N. S. and Gothilf, Y. (2006) Zebrafish arylalkyl-
27 Hardeland, R. and Pandi-Perumal, S. R. (2005) Melatonin, a amine-N-acetyltransferase genes – targets for regulation of
potent agent in antioxidative defense: actions as a natural the circadian clock. J. Mol. Endocrinol. 36, 337 – 347.
food constituent, gastrointestinal factor, drug and prodrug. 47 Gamse, J. T., Shen Y. C., Thisse, C., Thisse, B., Raymond, P.
Nutr. Metab. (Lond.) Sep. 10, 2 – 22. A., Halpern, M. E. and Liang, J. O. (2002) Otx5 regulates
28 Messner, M., Hardeland, R., Rodenbeck, A. and Huether, G. genes that show circadian expression in the zebrafish pineal
(1998) Tissue retention and subcellular distribution of con- complex. Nat. Genet. 30, 117 – 121.
tinuously infused melatonin in rats under near physiological 48 MuÇoz, E., Brewer, M. and Baler, R. (2006) Modulation of
conditions. J. Pineal Res. 25, 251 – 259. BMAL/CLOCK/E-Box complex activity by a CT-rich cis-
29 Tan, D.-X., Manchester, L. C., Reiter, R. J., Qi, W., Hanes, M. acting element. Mol. Cell. Endocrinol. 252, 74 – 81.
A. and Farley, N. J. (1999) High physiological levels of 49 Roseboom, P. H., Coon, S. L., Baler, R., McCune, S. K.,
melatonin in the bile of mammals. Life Sci. 65, 2523 – 2529. Weller, J. L. and Klein, D. C. (1996) Melatonin synthesis:
30 Bubenik, G. A., Hacker, R. R., Brown, G. M. and Bartos, L. analysis of the more than 150-fold nocturnal increase in
(1999) Melatonin concentrations in the luminal fluid, mucosa serotonin N-acetyltransferase messenger ribonucleic acid in
and muscularis of the bovine and porcine gastrointestinal the rat pineal gland. Endocrinology 137, 3033 – 3045.
tract. J. Pineal Res. 26, 56 – 63. 50 Koch, M., Mauhin, V., Stehle, J. H., Schomerus, C. and Korf,
31 Tan, D.-X., Manchester, L. C., Hardeland, R., Lopez-Burillo, H. W. (2003) Dephosphorylation of pCREB by protein serine/
S., Mayo, J. C., Sainz, R. M. and Reiter, R. J. (2003) Melatonin threonine phosphatases is involved in inactivation of Aanat
– a hormone, a tissue factor, an autocoid, a paracoid, and an gene transcription in rat pineal gland. J. Neurochem. 85, 170 –
antioxidant vitamin. J. Pineal Res. 34, 75 – 78. 179.
32 Huether, G., Poeggeler, B., Reimer, A. and George, A. (1992)
51 Karolczak, M., Korf, H. W. and Stehle, J. H. (2005) The
Effect of tryptophan administration on circulating melatonin
rhythm and blues of gene expression in the rodent pineal
levels in chicks and rats: evidence for stimulation of melatonin
gland. Endocrine 27, 89 – 100.
synthesis and release in the gastrointestinal tract. Life Sci. 51,
52 Ganguly, S., Gastel, J. A., Weller, J. L., Schwartz, C., Jaffe, H.,
945 – 953.
Namboodiri, M. A., Coon, S. L., Hickman, A. B., Rollag, M.,
33 Slominski, A., Fischer, T. W., Zmijewski, M. A., Wortsman, J.,
Obsil, T., Beauverger, P., Ferry, G., Boutin, J. A. and Klein, D.
Semak, I., Zbytek, B., Slominski, R. M. and Tobin, D. J. (2005)
C. (2001) Role of a pineal cAMP-operated arylalkylamine N-
On the role of melatonin in skin physiology and pathology.
acetyltransferase/14-3-3-binding switch in melatonin synthe-
Endocrine 27, 137 – 148.
sis. Proc. Natl. Acad. Sci. USA 98, 8083 – 8088.
34 Gaudet. S. J., Slominski, A., Etminan, M., Pruski, D., Paus, R.
and Namboodiri, M.A. (1993) Identification and character- 53 Choi, B. H., Chae, H. D., Park, T. J., Oh, J., Lim, J., Kang, S. S.,
ization of two isozymic forms of arylamine N-acetyltransfer- Ha, H. and Kim, K. T. (2004) Protein kinase C regulates the
ase in Syrian hamster skin. J. Invest. Dermatol. 101, 660 – 665. activity and stability of serotonin N-acetyltransferase. J.
35 Slominski, A., Pisarchik, A., Semak, I., Sweatman, T. and Neurochem. 90, 442 – 454.
Wortsman, J. (2003) Characterization of the serotoninergic 54 Schomerus, C., Korf, H. W., Laedtke, E., Weller, J. L. and
system in the C57BL/6 mouse skin. Eur. J. Biochem. 270, Klein, D. C. (2000) Selective adrenergic/cyclic AMP-depend-
3335 – 3344. ent switch-off of proteasomal proteolysis alone switches on
36 Fischer TW, Sweatman TW, Semak I, Sayre RM, Wortsman J, neural signal transduction: an example from the pineal gland.
Slominski A. (2006) Constitutive and UV-induced metabo- J. Neurochem. 75, 2123 – 2132.
lism of melatonin in keratinocytes and cell-free systems. 55 Stehle, J. H., von Gall, C. and Korf, H. W. (2001) Analysis of
FASEB J. 20, 1564 – 1566. Erratum in FASEB J. 21, 630. cell signalling in the rodent pineal gland deciphers regulators
37 Conti, A., Conconi, S., Hertens, E., Skwarlo-Sonta, K., of dynamic transcription in neural/endocrine cells. Eur. J.
Markowska, M. and Maestroni, G. J. M. (2000) Evidence for Neurosci. 14, 1 – 9.
melatonin synthesis in mouse and human bone marrow cells. J. 56 Kim, T. D., Kim J. S., Kim, J. H., Myung, J., Chae, H. D., Woo,
Pineal Res. 28, 193 – 202. K. C., Jang, S. K., Koh, D. S. and Kim, K. T. (2005) Rhythmic
38 Maestroni, G. J. M., Zammaretti, F. and Pedrinis, E. (1999) serotonin N-acetyltransferase mRNA degradation is essential
Hematopoietic effect of melatonin involvement of type 1 for the maintenance of its circadian oscillation. Mol. Cell.
kappa-opioid receptor on bone marrow macrophages and Biol. 25, 3232 – 3246.
interleukin-1. J. Pineal Res. 27, 145 – 153. 57 Iuvone, P. M., Tosini, G., Pozdeyev, N., Haque, R., Klein, D. C.
39 Hardeland, R. (1997) Melatonin: multiple functions in signal- and Chaurasia, S. S. (2005) Circadian clocks, clock networks,
ing and protection. In: Skin Cancer and UV Radiation, arylalkylamine N-acetyltransferase, and melatonin in the
pp. 186 – 198, Altmeyer, P., Hoffmann, K., and Stcker, M. retina. Prog. Retin. Eye Res. 24, 433 – 456.
(eds), Springer, Berlin 58 Chaurasia SS, Haque R, Pozdeyev N, Jackson CR, Iuvone PM.
40 Carrillo-Vico, A., Guerrero, J. M., Lardone, P. J. and Reiter, (2006) Temporal coupling of cyclic AMP and Ca/calmodulin-
R. J. (2005) A review of the multiple actions of melatonin on stimulated adenylyl cyclase to the circadian clock in chick
the immune system. Endocrine 27, 189 – 200. retinal photoreceptor cells. J. Neurochem. 99, 1142 – 1150.
41 Kvetnoy, I. M. (1999) Extrapineal melatonin: location and 59 Perreau-Lenz, S., Kalsbeek, A., Garidou, M. L., Wortel, J.,
role within diffuse neuroendocrine system. Histochem. J. 31, van der Vliet, J., van Heijningen, C., Simonneaux, V., Pvet, P.
1 – 12. and Buijs, R. M. (2003) Suprachiasmatic control of melatonin
42 Hardeland, R. (1999) Melatonin and 5-methoxytryptamine in synthesis in rats: inhibitory and stimulatory mechanisms. Eur.
non-metazoans. Reprod. Nutr. Dev. 39, 399 – 408. J. Neurosci. 17, 221 – 228.
2016 R. Hardeland Melatonin, hormone and more

60 Chik, C. L., Liu Q. Y., Li, B., Klein, D. C., Zylka, M., Kim, D. 76 Poirel, V. J., Cailotto, C., Streicher, D., Pvet, P., Masson-
S., Chin, H., Karpinski, E. and Ho, A. K. (1997) a1D L-type Pvet, M. and Gauer, F. (2003) MT1 melatonin receptor
Ca2+-channel currents: inhibition by a b-adrenergic agonist mRNA tissular localization by PCR amplification. Neuro-
and pituitary adenylate cyclase-activating polypeptide endocrinol. Lett. 24, 33 – 38.
(PACAP) in rat pinealocytes. J. Neurochem. 68, 1078 – 1087. 77 Brydon, L., Roka, F., Petit, L., de Coppet, P., Tissot, M.,
61 von Gall, C., Lewy, A., Schomerus, C., Vivien-Roels, B., Barrett, P., Morgan, P. J., Nanoff, C., Strosberg, A. D. and
Pevt, P., Korf, H. W. and Stehle, J. H. (2000) Transcription Jockers, R. (1999) Dual signaling of human Mel1a melatonin
factor dynamics and neuroendocrine signalling in the mouse receptors via Gi2, Gi3, and Gq/11 proteins. Mol. Endocrinol. 13,
pineal gland: a comparative analysis of melatonin-deficient 2025 – 2038.
C57BL mice and melatonin-proficient C3H mice. Eur. J. 78 Roy, D. and Belsham, D. D. (2002) Melatonin receptor
Neurosci. 12, 964 – 972. activation regulates GnRH gene expression and secretion in
62 Link, W. A., Ledo, F., Torres, B., Palczewska, M., Madsen, T. GT1 – 7 GnRH neurons: signal transduction mechanisms. J.
M., Savignac, M., Albar, J. P., Mellstrçm, B. and Naranjo, J. R. Biol. Chem. 277, 251 – 258.
(2004) Day-night changes in downstream regulatory element 79 Steffens, F., Zhou X.-B., Sausbier, U., Sailer, C., Motejlek, K.,
antagonist modulator/potassium channel interacting protein Ruth, P., Olcese, J., Korth, M. and Wieland, T. (2003)
activity contribute to circadian gene expression in pineal Melatonin receptor signaling in pregnant and nonpregnant
gland. J. Neurosci. 24, 5346 – 5355. rat uterine myocytes as probed by large conductance Ca2+-
63 Kim, T. D., Woo, K. C., Cho, S., Ha, D. C., Jang, S. K. and Kim, activated K+ channel activity. Mol. Endocrinol. 17, 2103 –
K. T. (2007) Rhythmic control of AANAT translation by 2115.
hnRNP Q in circadian melatonin production. Genes Dev. 21, 80 Chan, A. S., Lai, F. P., Lo, R. K., Voyno-Yasenetskaya, T. A.,
797 – 810. Stanbridge, E. J. and Wong, Y.H. (2002) Melatonin mt1 and
64 Ebisawa, T., Karne, S., Lerner, M. R. and Reppert, S. M. MT2 receptors stimulate c-Jun N-terminal kinase via pertussis
(1994) Expression cloning of a high-affinity melatonin toxin-sensitive and -insensitive G proteins. Cell Signal. 14,
receptor from Xenopus dermal melanophores. Proc. Natl. 249 – 257.
Acad. Sci. USA 91, 6133 – 6137. 81 Lai, F. P., Mody, S. M., Yung, L. Y., Kam, J. Y., Pang, C. S.,
65 Reppert, S. M., Weaver D. R. and Ebisawa, T. (1994) Cloning Pang, S. F., Wong, Y. H. (2002) Molecular determinants for the
and characterization of a mammalian melatonin receptor that differential coupling of Ga16 to the melatonin MT1, MT2 and
mediates reproductive and circadian responses. Neuron 13, Xenopus Mel1c receptors. J. Neurochem. 80, 736 – 745.
1177 – 1185. 82 Yung, L. Y., Tsim, S. T., Wong, Y. H. (1995) Stimulation of
66 Reppert, S. M., Godson, C., Mahle, C. D., Weaver, D. R., cAMP accumulation by the cloned Xenopus melatonin
Slaugenhaupt, S. A. and Gusella, J. F. (1995) Molecular receptor through Gi and Gz proteins. FEBS Lett. 372, 99 – 102.
characterization of a second melatonin receptor expressed in 83 McArthur, A. J., Hunt, E. A. and Gillette, M. U. (1997)
human retina and brain: the Mel1b melatonin receptor. Proc. Melatonin action and signal transduction in the rat supra-
Natl. Acad. Sci. USA 92, 8734 – 8738. chiasmatic circadian clock: activation of protein kinase C at
67 Jin, X., von Gall, C., Pieschl, R. L., Gribkoff, V. K., Stehle, J. dusk and dawn. Endocrinology 138, 627 – 634.
H., Reppert, S. M. and Weaver, D. R. (2003) Targeted 84 Godson, C. and Reppert, S. M. (1997) The Mel1a melatonin
disruption of the mouse Mel1b melatonin receptor. Mol. Cell. receptor is coupled to parallel signal transduction pathways.
Biol. 23, 1054 – 1060. Endocrinology 138, 397 – 404.
68 Dubocovich, M. L. and Markowska, M. (2005) Functional 85 Sharkey, J. and Olcese, J. (2007) Transcriptional inhibition of
MT1 and MT2 melatonin receptors in mammals. Endocrine 27, oxytocin receptor expression in human myometrial cells by
101 – 110. melatonin involves protein kinase C signaling. J. Clin.
69 Reppert, S. M., Weaver, D. R., Cassone, V. M., Godson, C. Endocrinol. Metab. 92, 4015 – 4019.
and Kolakowski, L.F. Jr. (1995) Melatonin receptors are for 86 Nosjean, O., Ferro, M., Cog, F., Beauverger, P., Henlin, J.-
the birds: molecular analysis of two receptor subtypes differ- M., Lefoulon, F., Fauch re, J.-L., Delagrange, P., Canet, E.
entially expressed in chick brain. Neuron 15, 1003 – 1015. and Boutin, J. A. (2000) Identification of the melatonin-
70 Jockers, R., Petit, L., Lacroix, I., de Coppet, P., Barrett, P., binding site MT3 as the quinone reductase 2. J. Biol.
Morgan, P. J., Guardiola, B., Delagrange, P., Marullo, S. and Chem. 275, 31311 – 31317.
Strosberg, A. D. (1997) Novel isoforms of Mel1c melatonin 87 Nosjean, O., Nicolas, J. P., Klupsch, F., Delagrange, P., Canet,
receptors modulating intracellular cyclic guanosine 3,5- E. and Boutin, J. A. (2001) Comparative parmacological
monophosphate levels. Mol. Endocrinol. 11, 1070 – 1081. studies of melatonin receptors: MT1, MT2 and MT3/QR2.
71 Hunt, A. E., Al-Ghoul, W. M., Gillette, M. U. and Duboco- Tissue distribution of MT3/QR2. Biochem. Pharmacol. 61,
vich, M. L. (2001) Activation of MT2 melatonin receptors in 1369 – 1379.
rat suprachiasmatic nucleus phase advances the circadian 88 Mailliet, F., Ferry, G., Vella, F., Berger, S., Cog, F.,
clock. Am. J. Physiol. 280, C110 – C118. Chomarat, P., Mallet, C., Gunin, S. P., Guillaumet, G.,
72 Liu, C., Weaver, D. R., Jin, X., Shearman, L. P., Pieschl, R. L., Viaud-Massuard, M. C., Yous, S., Delagrange, P. and Boutin,
Gribkoff, V. K. and Reppert, S. M. (1997) Molecular J. A. (2005) Characterization of the melatoninergic MT3
dissection of two distinct actions of melatonin on the supra- binding site on the NRH:quinone oxidoreductase 2 enzyme.
chiasmatic circadian clock. Neuron 19, 91 – 102. Biochem. Pharmacol. 71, 74 – 78.
73 Weaver, D. R., Liu, C., Reppert, S. M. (1996) Natures 89 Harada, S., Fujii, C., Hayashi, A. and Ohkoshi, N. (2001) An
knockout: the Mel1b receptor is not necessary for reproductive association between idiopathic Parkinsons disease and poly-
and circadian responses to melatonin in Siberian hamsters. morphisms of phase II detoxification enzymes: glutathione S-
Mol. Endocrinol. 10, 1478 – 1487. transferase M1 and quinone oxidoreductase 1 and 2. Biochem.
74 Schuster, C., Gauer, F., Guerrero, H., Lakhdar-Ghazal, N., Biophys. Res. Commun. 288, 887 – 892.
Pvet P. and Masson-Pvet, M. (2000) Photic regulation of 90 Long, D. J. II, Iskander, K., Gaikwad, A., Arin, M., Roop, D.
mt1 melatonin receptors in the Siberian hamster pars tuber- R., Knox, R., Barrios, R. amd Jaiswal, A. K. (2002) Disruption
alis and suprachiasmatic nuclei: involvement of the circadian of dihydronicotinamide riboside:quinone oxidoreductase 2
clock and intergeniculate leaflet. J. Neuroendocrinol. 12, (NQO2) leads to myeloid hyperplasia of bone marrow and
207 – 216. decreased sensitivity to menadione toxicity. J. Biol.
75 von Gall, C., Weaver, D. R., Moek, J., Jilg, A., Stehle, J. H. and Chem. 277, 46131 – 46139.
Korf, H. W. (2005) Melatonin plays a crucial role in the 91 Hardeland, R., Coto-Montes, A. and Poeggeler, B. (2003)
regulation of rhythmic clock gene expression in the mouse Circadian rhythms, oxidative stress and antioxidative defense
pars tuberalis. Ann. NY Acad. Sci. 1040, 508 – 511. mechanisms. Chronobiol. Int. 20, 921 – 962.
Cell. Mol. Life Sci. Vol. 65, 2008 Review Article 2017

92 Tan, D.-X., Manchester, L. C., Terron, M. P., Flores, L. J., 111 Tricoire, H., Locatelli, A., Chemineau, P. and Malpaux, B.
Tamura, H. and Reiter, R. J. (2007) Melatonin as a naturally (2002) Melatonin enters the cerebrospinal fluid through the
occurring co-substrate of quinone reductase-2, the putative pineal recess. Endocrinology 143, 84 – 90.
MT3 melatonin membrane receptor: hypothesis and signifi- 112 Tricoire, H., Møller, M., Chemineau, P. and Malpaux, B.
cance. J. Pineal Res. 43, 317 – 320. (2003) Origin of cerebrospinal fluid melatonin and possible
93 Ben
tez-King, G., Huerto-Delgadillo, L. and Antn-Tay, F. function in the integration of photoperiod. Reprod. Suppl.,
(1993) Binding of 3H-melatonin to calmodulin. Life Sci. 53, 61, 311 –321.
201 – 207. 113 Ma, X., Idle, J. R., Krausz, K. W., Tan, D.-X., Ceraulo, L. and
94 Ben
tez-King, G., R
os, A., Mart
nez, A. and Antn-Tay, F. Gonzalez, F. J. (2006) Urinary metabolites and antioxidant
(1996) In vitro inhibition of Ca2+/calmodulin-dependent products of exogenous melatonin in the mouse. J. Pineal
kinase II activity by melatonin. Biochim. Biophys. Acta Res. 40, 343 – 349.
1290, 191 – 196. 114 Tan, D.-X., Manchester L. C., Reiter, R. J., Plummer, B. F.,
95 Ben
tez-King G. (2006) Melatonin as a cytoskeletal modu- Hardies, L. J., Weintraub, S. T., Vijayalaxmi and Shepherd, A.
lator: implications for cell physiology and disease. J. Pineal M. (1998) A novel melatonin metabolite, cyclic 3-hydrox-
Res. 40, 1 – 9. ymelatonin: a biomarker of in vivo hydroxyl radical gener-
96 Hardeland, R. (1997) New actions of melatonin and their ation. Biochem. Biophys. Res. Commun. 253, 614 – 620.
relevance to biometeorology. Int. J. Biometeorol. 41, 47 – 57. 115 Tan, D.-X., Hardeland, R., Manchester, L. C., Poeggeler, B.,
97 Soto-Vega, E., Meza, I., Ram
rez-Rodr
guez, G. and Benitez- Lopez-Burillo, S., Mayo, J. C., Sainz, R. M. and Reiter, R. J.
King, G. (2004) Melatonin stimulates calmodulin phosphor- (2003) Mechanistic and comparative studies of melatonin and
ylation by protein kinase C. J. Pineal Res. 37, 98 – 106. classic antioxidants in terms of their interactions with the
98 Mac
as, M., Escames, G., Leon, J., Coto, A., Sbihi, Y., Osuna, ABTS cation radical. J. Pineal Res. 34, 249 – 259.
A. and AcuÇa-Castroviejo, D. (2003) Calreticulin — melato- 116 Hardeland, R., Poeggeler, B., Niebergall, R. and Zelosko, V.
nin. An unexpected relationship. Eur. J. Biochem. 270, 832 – (2003) Oxidation of melatonin by carbonate radicals and
840. chemiluminescence emitted during pyrrole ring cleavage. J.
99 Wiesenberg, I., Missbach, M., Kahlen J. P., Schrder, M. and Pineal Res. 34, 17 – 25.
Carlberg, C. (1995) Transcriptional activation of the nuclear 117 Tan, D.-X., Manchester L. C., Burkhardt, S., Sainz, R. M.,
receptor RZRa by the pineal gland hormone melatonin and Mayo, J. C., Kohen, R., Shohami E, Huo, Y.-S., Hardeland, R.
identification of CGP 52608 as a synthetic ligand. Nucleic and Reiter, R. J. (2001) N1-acetyl-N2-formyl-5-methoxyky-
Acids Res. 23, 327 – 333. nuramine, a biogenic amine and melatonin metabolite,
100 Carlberg, C. (2000) Gene regulation by melatonin. Ann. NY functions as a potent antioxidant. FASEB J. 15, 2294 – 2296.
Acad. Sci. 917, 387 – 396. 118 Tan, D.-X., Manchester, L. C., Di Mascio, P., Martinez, G. R.,
101 Toms-Zapico, C. and Coto-Montes, A. (2005) A proposed Prado, F. M. and Reiter, R. J. (2007) Novel rhythms of N1-
mechanism to explain the stimulatory effect of melatonin on acetyl-N2-formyl-5-methoxykynuramine and its precursor
antioxidative enzymes. J. Pineal Res. 39, 99 – 104. melatonin in water hyacinth: importance for phytoremedia-
102 Pandi-Perumal, S. R., Srinivasan, V., Maestroni, G. J. M., tion. FASEB J. 21, 1724 – 1729.
Cardinali, D. P., Poeggeler, B. and Hardeland, R. (2006) 119 Tan, D.-X., Reiter, R. J., Manchester, L. C., Yan, M.-T., El-
Melatonin: Natures most versatile biological signal? FEBS J. Sawi, M., Sainz, R. M., Mayo, J. C., Kohen, R., Allegra, M.
273, 2813 – 2838. and Hardeland, R. (2002) Chemical and physical properties
103 Andrabi, S. A., Sayeed, I., Siemen, D., Wolf, G. and Horn, T. F. and potential mechanisms: melatonin as a broad spectrum
(2004) Direct inhibition of the mitochondrial permeability antioxidant and free radical scavenger. Curr. Top. Med.
transition pore: a possible mechanism responsible for anti- Chem. 2, 181 – 197.
apoptotic effects of melatonin. FASEB J. 18, 869 – 871. 120 Agozzino, P., Avellone, G., Bongiorno, D., Ceraulo, L.,
104 Reiter, R. J., Tan, D.-X., Mayo, J. C., Sainz, R. M., Leon, J. and Filizzola, F., Natoli, M. C., Livrea, M. A. and Tesoriere, L.
Czarnocki, Z. (2003) Melatonin as an antioxidant: biochem- (2003) Melatonin: structural characterization of its non-
ical mechanisms and pathophysiological implications in enzymatic mono-oxygenate metabolite. J. Pineal Res. 35,
humans. Acta Biochim. Pol. 50, 1129 – 1146. 269 – 275.
105 Hardeland, R. (2005) Antioxidative protection by melatonin: 121 Tan, D.-X., Manchester, L. C. Reiter, R. J., Qi, W.-B.
multiplicity of mechanisms from radical detoxification to Karbownik, M. and Calvo, J. R. (2000) Significance of
radical avoidance. Endocrine 27, 119 – 130. melatonin in antioxidative defense system: reactions and
106 Menendez-Pelaez, A., Poeggeler, B., Reiter, R. J., Barlow- products. Biol. Signals Recept. 9, 137 – 159.
Walden, L., Pablos, M. I. and Tan D.-X. (1993) Nuclear 122 Seever, K. and Hardeland, R. (2008) Novel pathway for N1-
localization of melatonin in different mammalian tissues: acetyl-5-methoxykynuramine: UVB-induced liberation of
immunocytochemical and radioimmunoassay evidence. J. carbon monoxide from precursor N1-acetyl-N2-formyl-5-me-
Cell. Biochem. 53, 373 – 382. thoxykynuramine. J. Pineal Res., in press.
107 Mondaca, M., Hernndez, A., Valladares, L., Sierralta, W., 123 Rosen, J., Than, N. N. Koch, D., Poeggeler, B., Laatsch, H. and
Noseda, R. and Soto-Moyano, R. (2004) Involvement of Hardeland, R. (2006) Interactions of melatonin and its
melatonin metabolites in the long-term inhibitory effect of the metabolites with the ABTS cation radical: extension of the
hormone on rat spinal nociceptive transmission. Pharmacol, radical scavenger cascade and formation of a novel class of
Biochem. Behav. 77, 275 – 279. oxidation products, C2-substituted 3-indolinones. J. Pineal
108 Teh, M. T. and Sugden, D. (2002) Desensitization of pigment Res. 41, 374 – 381.
granule aggregation in Xenopus leavis melanophores: mela- 124 Ressmeyer, A.-R., Mayo, J. C. Zelosko, V., Sinz, R. M. Tan,
tonin degradation rather than receptor down-regulation is D.-X., Poeggeler, B., Antol
n, I., Zsizsik, B. K. Reiter, R. J.
responsible. J. Neurochem. 81, 719 – 727. and Hardeland, R. (2003) Antioxidant properties of the
109 Ferry, G., Ubeaud, C., Lambert, P. H., Bertin, S., Cog, F., melatonin metabolite N1-acetyl-5-methoxykynuramine
Chomarat, P., Delagrange, P., Serkiz, B., Bouchet, J. P., (AMK): scavenging of free radicals and prevention of protein
Truscott, R. J. and Boutin, J. A. (2005) Molecular evidence destruction. Redox Rep. 8, 205 – 213.
that melatonin is enzymatically oxidized in a different manner 125 Guenther, A.L., Schmidt, S. I., Laatsch, H., Fotso, S., Ness,
than tryptophan: investigation on both indoleamine-2,3- H., Ressmeyer, A.-R., Poeggeler, B. and Hardeland, R.
dioxygenase and myeloperoxidase. Biochem. J. 388, 205 – 215. (2005) Reactions of the melatonin metabolite AMK (N1-
110 Ma, X., Idle, J. R., Krausz, K. W. and Gonzalez, F. J. (2005) acetyl-5-methoxykynuramine) with reactive nitrogen species:
Metabolism of melatonin by human cytochromes p450. Drug formation of novel compounds, 3-acetamidomethyl-6-meth-
Metab. Dispos. 33, 489 – 494. ACHTUNGREoxycinnolinone and 3-nitro-AMK. J. Pineal Res. 39, 251 – 260.
2018 R. Hardeland Melatonin, hormone and more

126 Hardeland, R., Backhaus, C. and Fadavi, A. (2007) Reactions effect of PGE2 on IL-2 production in human lymphocytes via
of the NO redox forms NO+, ·NO and HNO (protonated its mt1 membrane receptor. FASEB J. 17, 755 – 757.
NO) with the melatonin metabolite N1-acetyl-5-methoxyky- 145 Lotufo, C. M., Lopes, C., Dubocovich, M. L., Farsky, S. H. and
nuramine (AMK). J. Pineal Res. 43, 382 – 388. Markus, R. P. (2001) Melatonin and N-acetylserotonin inhibit
127 Hardeland, R., Backhaus, C., Fadavi, A. and Hess, M. (2007) leukocyte rolling and adhesion to rat microcirculation. Eur. J.
N1-acetyl-5-methoxykynuramine contrasts with other trypto- Pharmacol. 430, 351 – 357.
phan metabolites by a peculiar type of NO scavenging: 146 Markowska, M., Mrozkowiak, A., Pawlak, J. and Skwarło-
cyclization to a cinnolinone prevents formation of unstable Sońta, K. (2004) Intracellular second messengers involved in
nitrosamines. J. Pineal Res. 43, 104 – 105. melatonin signal transduction in chicken splenocytes in vitro.
128 Tan, D.-X., Chen, L.-D., Poeggeler, B., Manchester, L. C. and J. Pineal Res. 37, 207 – 212.
Reiter, R. J. (1993) Melatonin: a potent, endogenous hydroxyl 147 Carrillo-Vico, A., Garc
a-PergaÇeda, A., Naji, L., Calvo, J.
radical scavenger. Endocr. J. 1, 57 – 60. R., Romero, M. P. and Guerrero, J. M. (2003) Expression of
129 Mart
n, M., Mac
as, M., Escames, G., Reiter, R. J., Agapito, membrane and nuclear melatonin receptor mRNA and
M. T., Ortiz, G. G. and AcuÇa-Castroviejo, D. (2000) protein in the mouse immune system. Cell. Mol. Life Sci. 60,
Melatonin-induced increased activity of the respiratory 2272 – 2278.
chain complexes I and IV can prevent mitochondrial damage 148 Balzer, I. and Hardeland, R. (1989) Action of kynuramine in a
induced by ruthenium red in vivo. J. Pineal Res. 28, 242 – 248. dinoflagellate: stimulation of bioluminescence in Gonyaulax
130 AcuÇa-Castroviejo, D., Escames, G., Len, J., Carazo, A. and polyedra. Comp. Biochem. Physiol. 94C, 129 – 132.
Khaldy, H. (2003) Mitochondrial regulation by melatonin and 149 Kennaway, D. J., Blake, P. and Webb, H. A. (1989) A
its metabolites. Adv. Exp. Med. Biol. 527, 549 – 557. melatonin agonist and N-acetyl-N2-formyl-5-methoxykynur-
131 Okatani, Y., Wakatsuki, A. and Reiter, R. J. (2002) Melatonin enamine accelerate the reentrainment of the melatonin
protects hepatic respiratory chain activity in senescence- rhythm following a phase advance of the light-dark cycle.
accelerated mice. J. Pineal Res. 32, 143 – 148. Brain Res. 495, 349 – 354.
132 Okatani, Y., Wakatsuki, A., Reiter, R. J. and Miyahara, Y. 150 Budu, A., Peres, R., Bueno, V. B., Catalani, L. H. and da Silva
(2002) Hepatic mitochondrial dysfunction in senescence-accel- Garcia, C. R. (2007) N1-acetyl-N2-formyl-5-methoxykynur-
erated mice: correction by long-term, orally administered amine modulates the cell cycle of malaria parasites. J. Pineal
physiological levels of melatonin. J. Pineal Res. 33, 127 – 133. Res. 42, 261 – 266.
133 Baydas, G., Reiter, R. J., Akbulut, M., Tuzcu, M. and Tamer, 151 Kelly, R. W., Amato, F. and Seamark, R. F. (1984) N-Acetyl-5-
S. (2005) Melatonin inhibits neural apoptosis induced by methoxy kynurenamine, a brain metabolite of melatonin, is a
homocysteine in hippocampus of rats via inhibition of potent inhibitor of prostaglandin biosynthesis. Biochem.
cytochrome c translocation and caspase-3 activation and by Biophys. Res. Commun. 121, 372 – 379.
regulating pro- and anti-apoptotic protein levels. Neuro- 152 Mayo, J. C., Sainz R. M., Tan, D.-X., Hardeland, R., Leon, J.,
science 135, 879 – 886. Rodriguez, C. and Reiter, R. J. (2005) Anti-inflammatory
134 Sourdeval, M., Lemaire, C., Deniaud, A., Taysse, L., Daulon, actions of melatonin and its metabolites, N1-acetyl-N2-
S., Breton, P., Brenner, C., Boisvieux-Ulrich, E. and Marano, formyl-5-methoxykynuramine (AFMK) and N1-acetyl-5-me-
F. (2006) Inhibition of caspase-dependent mitochondrial thoxykynuramine (AMK), in macrophages. J. Neuroimmu-
permeability transition protects airway epithelial cells against nol. 165, 139 – 149.
mustard-induced apoptosis. Apoptosis 11, 1545 – 1559. 153 Silva, S. O., Ximenes, V. F., Livramento, J. A., Catalani, L. H.
135 Xu, M. and Ashraf, M. (2002) Melatonin protection against and Campa, A. (2005) High concentrations of the melatonin
lethal myocyte injury induced by doxorubicin as reflected by metabolite, N1-acetyl-N2-formyl-5-methoxykynuramine, in
effects on mitochondrial membrane potential. J. Mol. Cell. cerebrospinal fluid of patients with meningitis: a possible
Cardiol. 34, 75 – 79. immunomodulatory mechanism. J. Pineal Res. 39, 302 – 306.
136 Prada, C., Udin, S. B., Wiechmann, A. F. and Zhdanova, I. V. 154 Silva, S. O., Rodrigues, M. R., Ximenes V. F., Bueno-da-Silva,
(2005) Stimulation of melatonin receptors decreases calcium A. E., Amarante-Mendes, G. P. and Campa, A. (2004)
levels in Xenopus tectal cells by activating GABAC receptors. Neutrophils as a specific target for melatonin and kynur-
J. Neurophysiol. 94, 968 – 978. amines: effects on cytokine release. J. Neuroimmunol. 156,
137 Prada, C. and Udin, S. B. (2005) Melatonin decreases calcium 146 – 152.
levels in retinotectal axons of Xenopus laevis by indirect 155 Burkhardt, S., Reiter R. J., Tan, D.-X., Hardeland, R.,
activation of group III metabotropic glutamate receptors. Cabrera, J. and Karbownik, M. (2001) DNA oxidatively
Brain Res. 1053, 67 – 76. damaged by chromium(III) and H2O2 is protected by the
138 Liu, L. Y., Hoffman, G. E., Fei, X. W., Li, Z., Zhang, Z. H. and antioxidants melatonin, N1-acetyl-N2-formyl-5-methoxyky-
Mei, Y. A. (2007) Delayed rectifier outward K+ current nuramine, resveratrol and uric acid. Int. J. Biochem. Cell
mediates the migration of rat cerebellar granule cells stimu- Biol. 33, 775 – 783.
lated by melatonin. J. Neurochem. 102, 333 – 344. 156 Onuki, J., Almeida, E. A., Medeiros, M. H. G. and Di Mascio,
139 Zhang, M., Cao, L. H., Yang, X. L. (2007) Melatonin P. (2005) Inhibition of 5-aminolevulinic acid-induced DNA
modulates glycine currents of retinal ganglion cells in rat. damage by melatonin, N1-acetyl-N2-formyl-5-methoxykynur-
Neuroreport 18, 1675 – 1678. amine, quercetin or resveratrol. J. Pineal Res. 38, 107 – 115.
140 Lardone, P. J., Carrillo-Vico, A., Naranjo, M. C., De Felipe, 157 Manda, K., Ueno, M. and Anzai, K. (2007) AFMK, a
B., Vallejo, A., Karasek, M. and Guerrero, J. M. (2006) melatonin metabolite, attenuates X-ray-induced oxidative
Melatonin synthesized by Jurkat human leukemic T cell line is damage to DNA, proteins and lipids in mice. J. Pineal Res. 42,
implicated in IL-2 production. J. Cell Physiol. 206, 273 – 279. 386 – 393.
141 Guerrero, J. M. and Reiter, R. J. (2002) Melatonin-immune 158 Len, J., Escames, G., Rodr
guez, M. I., Lpez, L. C., Tapias,
system relationships. Curr. Top. Med. Chem. 2, 167 – 179. V., Entrena, A., Camacho, E., Carrin, M. D., Gallo, M. A.,
142 Carrillo-Vico, A., Reiter R. J., Lardone, P. J., Herrera, J. L., Espinosa, A., Tan, D.-X., Reiter, R. J. and AcuÇa-Castroviejo,
Fernndez-Montesinos, R., Guerrero, J. M. and Pozo, D. D. (2006) Inhibition of neuronal nitric oxide synthase activity
(2006) The modulatory role of melatonin on immune by N1-acetyl-5-methoxykynuramine, a brain metabolite of
responsiveness. Curr. Opin. Invest. Drugs 7, 423 – 431. melatonin. J. Neurochem. 98, 2023 – 2033.
143 Garc
a-MauriÇo, S., Pozo, D., Carrillo-Vico, A., Calvo, J. R.
and Guerrero, J. M. (1999) Melatonin activates Th1 lympho-
cytes by increasing IL-12 production. Life Sci. 65, 2143 – 2150.
144 Carrillo-Vico, A., Garc
a-MauriÇo, S., Calvo, J. R. and
Guerrero, J. M. (2003) Melatonin counteracts the inhibitory