Fcell 08 00791

REVIEW
published: 21 August 2020

doi: 10.3389/fcell.2020.00791
From Sperm Motility to Sperm-Borne

microRNA Signatures: New
Approaches to Predict Male Fertility
Potential
Maíra Bianchi Rodrigues Alves 1,2 , Eneiva Carla Carvalho Celeghini 2 and
Clémence Belleannée 1*
1
CHU de Québec Research Center (CHUL), Department of Obstetrics, Gynecology and Reproduction, Faculty of Medicine,
Université Laval, Quebec City, QC, Canada, 2 Department of Animal Reproduction, Universidade de São Paulo,
Pirassununga, Brazil
In addition to the paternal genome, spermatozoa carry several intrinsic factors,

including organelles (e.g., centrioles and mitochondria) and molecules (e.g., proteins
and RNAs), which are involved in important steps of reproductive biology such as
spermatogenesis, sperm maturation, oocyte fertilization and embryo development.
These factors constitute potential biomarkers of “viable sperm” and male fertility status
Edited by: and may become major assets for diagnosing instances of idiopathic male infertility
Tzviya Zeev-Ben-Mordehai,
Utrecht University, Netherlands
in both humans and livestock animals. A better understanding of the mechanism
Reviewed by:
of action of these sperm intrinsic factors in the regulation of reproductive and
Myung-Geol Pang, developmental processes still presents a major challenge that must be addressed.
Chung-Ang University, South Korea
This review assembles the main data regarding morpho-functional and intrinsic sperm
Brett Nixon,
The University of Newcastle, Australia features that are associated with male infertility, with a particular focus on microRNA
*Correspondence: (miRNA) molecules.
Clémence Belleannée
Keywords: infertility, diagnosis, spermatozoa, semen, intrinsic factors, ncRNAs, human, cattle
Clemence.Belleannee@
crchudequebec.ulaval.ca
Specialty section: INTRODUCTION

This article was submitted to
Cell Growth and Division, Male fertility potential relies on several physical, endocrine, and genetic factors, which underlie
a section of the journal the production of fully functional spermatozoa as well as their successful arrival at the site of
Frontiers in Cell and Developmental fertilization. Impairment of these male reproductive functions accounts for half of the infertility
Biology issues that couples face during their reproductive age (Agarwal et al., 2015; Zegers-Hochschild
Received: 26 May 2020 et al., 2017). While male infertility diagnosis is mainly based on physical characteristics, hormonal
Accepted: 28 July 2020 analysis, and use of the traditional spermogram, its etiology remains unexplained in more than 30%
Published: 21 August 2020
of patients (Irvine, 1998; Agarwal et al., 2015). Thus, the design of novel customized diagnostic tools
Citation: to fully assess male fertility potential and male infertility etiology is a necessity. Furthermore, an
Alves MBR, Celeghini ECC and improved method to assess male fertility potential would be a major asset to the livestock industry,
Belleannée C (2020) From Sperm
since this parameter constitutes a serious economic challenge in this field. Research performed
Motility to Sperm-Borne microRNA
Signatures: New Approaches
on sperm samples from humans, livestock animals and rodents recently questioned the definition
to Predict Male Fertility Potential. of the sperm cell as a simple carrier of the paternal genome. From this, it now appears that
Front. Cell Dev. Biol. 8:791. the spermatozoon is a well-differentiated cell that carries a widely diverse signature of specific
doi: 10.3389/fcell.2020.00791 organelles and molecules, that could vary according to male fertility potential, and be transferred to
Frontiers in Cell and Developmental Biology | www.frontiersin.org 1 August 2020 | Volume 8 | Article 791
Alves et al. Sperm Factors and Male Infertility
the oocyte at the time of fertilization. This contemporary view zona pellucida, the previous paradigm supporting the role of
of the sperm cell opens potential new avenues concerning the enzymes released during acrosome reaction in the digestion
development of a novel generation of diagnostic tools. In this of the zona pellucida has been revisited in different species
review, we will delineate structural, morpho-functional and (Hirohashi and Yanagimachi, 2018).
intrinsic sperm features, and comment on their contribution The sperm tail (or flagellum) includes the neck, midpiece,
to the post-fertilization processes. The detection of sperm principal piece and terminal piece. The sperm centrioles are
microRNAs (miRNAs) as a predictor of male fertility potential important to early embryo development and are localized in the
will be discussed with regard to its possible applications in both sperm neck (Avidor-Reiss and Fishman, 2019). The axoneme,
clinic and industry. located internally along the entire flagellum, is composed of
nine peripheral doublets and two central single microtubules
(9 + 2 structure) integrated by the intraflagellar transport
“HEALTHY SPERM” CONCEPT: (IFT) system. Surrounding the axoneme, there is the outer
STRUCTURAL, MORPHO-FUNCTIONAL dense fibers (ODF) and mitochondria in the sperm midpiece
AND INTRINSIC FEATURES and the fibrous sheath (FS), formed by nine bundles of
fibers of different lengths, in the principal piece (Inaba, 2003).
While sperm quality is a key determinant of male fertility Depending on species, approximately 22–75 mitochondria are
potential, the discrimination between a high- and low- present in the midpiece to produce enough energy necessary
quality sperm sample is challenging. Indeed, this relies on a for a spermatozoon to transit along the female reproductive
broad spectrum of sperm features (Amann and Hammerstedt, tract and to reach the fertilization site in the oviduct (Song
1993). According to this concept, “healthy spermatozoa” and Lewis, 2008). All of these sperm structural characteristics
also referred to as “viable spermatozoa” should possess the are essential to ensure the ability of spermatozoa to cross
ability to reach the fertilization site, bind to and fertilize the muco-cervical and uterine barriers and reach the oviduct,
the oocytes, and properly contribute to initiation of early where they bind to and penetrate the oocyte to deliver
embryo development (Krawetz, 2005). These abilities are their DNA content.
strictly dependent on specific structural, morpho-functional, and
intrinsic features (Figure 1). Morpho-Functional Sperm Features
Morpho-functional sperm attributes are capable of modulating
Structural Sperm Features male fertility potential and are tightly related to the structural
Spermatozoa are the most differentiated cells of the organism sperm features. Parameters such as sperm motility/kinetics,
and display particular features related to their main function: the morphological abnormalities, integrity of plasma and
delivery of paternal DNA to the oocyte (Krawetz, 2005). In this acrosome membranes, mitochondrial activity production
regard, spermatozoa are composed of two main parts: a head to of reactive oxygen species (ROS), DNA fragmentation
carry and protect the genome, and a tail or flagellum to propel the and capacitation status are included in this group and
cell to the site of fertilization (Figure 2). usually are associated with male fertility (Table 1 and
Within the sperm head is a limited quantity of cytoplasm, Figure 3). Thus, assessment of these parameters is essential
highly condensed DNA, and a well-delimited acrosome. to determine male fertility potential: evaluation of a
A substantial portion of the cytoplasm is lost during the final higher number of morpho-functional attributes establishes
steps of spermatogenesis, specifically during spermiogenesis, the highest relationship with male fertility (Amann and
the process during which round spermatids differentiate into Hammerstedt, 1993). A high proportion of morphological
elongated spermatids and then spermatozoa (Gadea et al., abnormalities – referred to as teratozoospermia – including,
2013). The remaining cytoplasm from intercellular bridges, e.g., spermatozoa with large, small or piriform heads as well
called the cytoplasmic droplet, is lost during sperm transit as coiled-tails, is associated with reproductive dysfunctions
through the epididymis (Cooper and Yeung, 2003). During (Gillan et al., 2008). This common cause of male infertility is
spermiogenesis, the sequential replacement of histones by routinely assessed by light microscopic analysis of semen in
transitional proteins and then by protamines within sperm fertility clinics.
chromatin triggers genome condensation (Zini and Agarwal, Several other techniques are currently available to assess
2011; Gadea et al., 2013). The acrosome, located at the top sperm morpho-functional attributes. Sperm motility,
of the head, contains specific enzymes that promote specific kinetics, vigor and hyperactivation levels, are morpho-
functions trailing sperm capacitation and acrosome reaction: (1) functional features classically assessed by light microscopy
exposure of acrosome zona pellucida binding proteins during or computer-assisted sperm analysis (i.e., CASA) (Vincent
sperm capacitation; (2) sperm ability to cross cumulus cells et al., 2012). In addition, since sperm motility requires a
that surrounds the oocytes, (3) sperm binding to zona pellucida substantial amount of energy produced by mitochondrial
and acrosome reaction, and (4) migration of IZUMO1 protein activity (Piomboni et al., 2012), high fertility samples usually
from the outer acrosomal membrane to the equatorial segment present with high mitochondrial membrane potential. This
of sperm surface to ensure its binding to JUNO receptor on feature can be assessed by fluorescence microscopy and flow
the oocyte (Satouh et al., 2012; Bianchi et al., 2014). Since cytometric approaches mainly using the JC-1 fluorescent
acrosome reacted sperm retain the ability to penetrate the probe, which forms J-aggregates when mitochondria are active
FIGURE 1 | Concept of “healthy (viable) sperm.” Male fertility potential relies on structural, morpho-functional, and intrinsic sperm features that shape equally the
concept of “healthy (viable) sperm.”
and polarized; in this situation, more JC-1 monomers enter is proposed to predict litter size and fertility success in pigs
the spermatozoa and aggregate to generate red fluorescence (Kwon et al., 2015).
(Gillan et al., 2005; Celeghini et al., 2007). In addition to While the assessment of these features is essential to determine
energy production, mitochondria also produce large quantities male fertility potential, based on data from human clinics, about
of ROS that promote oxidative stress and damage to sperm 30% of sperm samples presenting with satisfactory morpho-
membranes and DNA. Therefore, ROS quantification functional features (Figure 3) are still unable to fertilize
in sperm cells is also an important morpho-functional the oocytes or to trigger early embryo development. Recent
feature that can be assessed by fluorescence microscopy, advances in analyses of sperm molecular and subcellular content
flow cytometry and molecular/biochemical assays using suggested that intrinsic factors may participate to early embryo
fluorescent probes. Such assays include BODIPY, which development and, consequently, could be potential targets for the
measures the susceptibility of sperm to lipid peroxidation, assessment of male fertility potential.
CellROX Deep Red , which stains ROS present in sperm
R
cells, and measurement of malondialdehyde concentration Intrinsic Sperm Features

(produced during lipid peroxidation) using the TBARS In addition to the paternal DNA, the spermatozoon carries
(Thiobarbituric acid reactive substances) assay (Aitken numerous molecules and organelles, which constitute a growing
et al., 2007; Celeghini et al., 2019). In addition, since focus of interest in fundamental research. Among these intrinsic
damage to the sperm membranes and DNA impairs sperm factors, sperm centrioles, sperm mitochondria, sperm proteins,
capacitation and early embryo development, both plasma and sperm RNAs are particularly interesting with respect to their
and acrosome membrane as well as DNA integrity should potential contribution to the post-fertilization process (Figure 4).
be evaluated by fluorescence microscopy or flow cytometric As shown in Figure 2, sperm centrioles are localized in the
approaches. Propidium iodide fluorescent probes and sperm neck while sperm mitochondria are arranged in the
fluorescent lectin from Pisum sativum can be used to assess midpiece. Since mature spermatozoa possess a minimal amount
plasma and acrosome membrane damage (Celeghini et al., of cytoplasm and a nucleus with highly condensed DNA that
2007), respectively, as well as SCSA (Sperm Chromatin R
prevents transcription, the presence of proteins and RNAs in
Structure Assay) to detect the susceptibility of sperm to sperm cells is limited. However, several proteins and ribonucleic
DNA fragmentation (Evenson and Wixon, 2006). Post-sperm acid molecules are likely acquired by the maturing spermatozoa
capacitation status could be assessed using the combination during spermatogenesis and their transit through the epididymis
of Hoechst 33258/chlortetracycline fluorescence probes and and post-ejaculatory journey (Boerke et al., 2007).
result of their acquisition by sperm during passage through the

epididymis and by the contact with the fluids coming from the
accessory sexual glands (e.g., clusterin encoding RNA, Hermo
et al., 1994); and (4) non-coding RNAs (ncRNAs) that are
acquired during the late steps of spermatogenesis or during
post-testicular sperm maturation with a potential function at the
time of fertilization (e.g., miRNAs and tRNA derived fragments,
Sharma et al., 2016; Yuan et al., 2016). Among RNA species
present in spermatozoa, ribosomal RNAs and mitochondrial
RNAs (mtRNAs) are the most abundant, followed by ncRNAs.
In contrast to somatic cells that comprise approximately 10 pg of
RNA, mature spermatozoa carry around a 1,000-fold lower RNA
content (Odia et al., 2018).
In addition to mRNAs, other sperm-borne RNA species,
including miRNAs have been associated with sperm cell fertility
status and early embryo development (Liu et al., 2012). The
central dogma of molecular biology supports that genes are
transcribed in the form of mRNAs, which in turn are translated
into proteins, with the exception of structural RNAs such as
ribosomal RNA (Mattick, 2001). Advancements in molecular
biology and studies on the human genome have revealed that
only 2% of all genes actually encode proteins (Mattick, 2001). In
addition, many genomic sequences that were first described as
“junk DNA” due to the fact that they did not encode proteins and
had no apparent function, now appear to play an important role
as post-transcriptional modulators. In this context, a large group
of RNAs called non-coding RNAs (ncRNAs) has been identified
in spermatozoa, including miRNAs (Wightman et al., 1993).
MicroRNAs are ∼18–22-nucleotide sequences that modulate
FIGURE 2 | Structural sperm features. Spermatozoa are composed of two
gene expression following their pairing with the untranslated
main parts: head and tail (or flagellum). The sperm head is constituted
basically by the acrosome and nucleus. The sperm tail includes: the neck that
region (30 UTR) of target transcripts (Ambros, 2004; Bartel,
contains mainly the proximal centriole; the midpiece which is composed by 2004). The biosynthesis of miRNAs is performed in the cell
mitochondria, outer dense fibers (ODF) and axoneme; principal piece nucleus with the generation of primary miRNA transcripts (pri-
containing the fibrous sheath and axoneme; and terminal piece. miRNA) and then precursor miRNAs (pre-miRNA), which are
transported to the cytoplasm. In the cytoplasm, the hairpin
structure is cleaved by Dicer, resulting in small double-stranded
transcripts. The miRNA double strand is then transferred to
RNAS SPECIES ARE MAJOR SPERM
Argonaut proteins, which bind to the mature miRNA also called
INTRINSIC FACTORS guide-miRNA. This complex promotes gene silencing through
translational inhibition or cleavage of mRNA (Ambros, 2004;
microRNAs and Other Sperm RNA Bartel, 2004). Such cleavage is common in plants and occurs
Species when the pairing between the miRNA and 30 UTR region is
Sperm RNAs were first described in the 1970s in murine perfect. On the other hand, an imperfect pairing implies the
and bovine spermatozoa (Betlach and Erickson, 1973; Paul blockade of translation machinery on that particular mRNA
and Duerksen, 1975). Given the limited quantity of cytoplasm without degradation—the most common process in animals
and high level of sperm DNA condensation, this finding has (Bartel, 2004, 2018).
been questioned as RNAs were potentially providing from Of interest is that miRNAs either act in the cell where they
mitochondria or contamination from somatic cells (Krawetz, are transcribed or in target cells through an intercellular
2005). In that regards, the roles and the origin of sperm RNAs communication mechanism (Raposo and Stoorvogel,
remain to be determined (Ostermeier et al., 2004; Krawetz, 2005). 2013). Extracellular vesicles are important mediators of
Four different RNA species can be distinguished in this communication. Exosomes comprise a population
spermatozoa (Boerke et al., 2007): (1) mRNAs that are remnants of small (40–160 nm) extracellular vesicles derived from
from the spermatogenesis process with no known function in intracellular multivesicular bodies carrying diverse bioactive
the oocyte (e.g., protamine-2 encoding RNA, Ziyyat, 2001); (2) molecules, including miRNAs, mRNAs and proteins (Raposo
mRNAs that originate from spermatogenesis with a potential and Stoorvogel, 2013). Once exosomes are released into
function in the oocytes (e.g., PLCζ encoding RNA, Saunders the extracellular environment, they can be internalized
et al., 2002); (3) mRNAs classified as “foreign” RNAs as a by recipient target cells (Raposo and Stoorvogel, 2013).
TABLE 1 | Correlation between sperm morpho-functional attributes and male fertility.
Sperm Definition Required equipment Correlation with fertility References

morpho-functional
attribute
Sperm subjective motility Maximum percentage of sperm Light field optical microscope, r = 0.53 (P < 0.01) (1 60 to 90-day Correa et al., 1997
cells with movement preferably with phase contrast non-return)
r = 0.67 (P = 0.03) (1 56-day non-return) Gillan et al., 2008
Computer assisted sperm Percentage of sperm with Light field optical microscope, WOB2 : r = 0.57 (P < 0.05) (1 56-day Morrell et al., 2018
analysis (CASA) motility and measurement of with phase contrast coupled to non-return)
different sperm kinetic features a computer
r = 0.67 (P = 0.03) (1 56-day non-return) Gillan et al., 2008
VSL3 : r = −0.34 (P < 0.05) (1 56-day Sellem et al., 2015
non-return)
ALH4 and progressive motility Farrell et al., 1998
presented r 2 of 0.68
ALH4 , BCF5 , LIN6 , VAP7 and VSL3 Farrell et al., 1998
presented r 2 of 0.98
Sperm morphological Percentage of cells that present Light field optical microscope r = −0.59 (P = 0.01) (1 60 to 90-day Correa et al., 1997
abnormalities with form defects, often called preferably with differential non-return)
abnormal cell percentage interference contrast
(Blom, 1973)
r = −0.76 (P < 0.05) (1 56-day Gillan et al., 2008
non-return)
r = −0.62 (P < 0.05) (1 56-day Morrell et al., 2018
non-return)
IPIAH8 Percentage of IPIAH8 sperm Fluorescence microscopy or High IPIAH8 (44.5%): 64.7% of Oliveira et al., 2014
flow cytometry pregnancy rate9 Low IPIAH8 (8.5%):
36.2% of pregnancy rate9
ROS Percentage of sperm cells Fluorescence microscopy or High ROS (10.63%): low fertility rates Celeghini et al., 2019
producing ROS flow cytometry Low ROS (6.11%): high fertility rates
DNA fragmentation Percentage of sperm cells with Fluorescence microscopy or r = −0.56 (P < 0.01) Morrell et al., 2018
DNA fragmentation flow cytometry
Capacitation status Percentage of Fluorescence microscopy or r = 0.37 (P < 0.05) (1 litter size) Kwon et al., 2015
acrosome-reacted flow cytometry
spermatozoa
1 Fertility measurement; 2 wobble (%); 3 progressive velocity (µm/s); 4 lateral head amplitude (µm); 5 beat cross frequency (Hz); 6 linearity (%); 7 path velocity (µm/s);
8 spermatozoa with intact plasma and acrosome membranes and high mitochondrial membrane potential; 9 ultrassonography exam 30 days after insemination.
Exosomes are released from different internal organs of are not only responsible for transporting genetic material, but
the male reproductive tract, including the epididymis also as contributors to the activation of the oocyte and the
(epididymosomes) and prostate (prostasomes) (Sullivan cellular structure and molecular components of the zygote as
and Saez, 2013). Epididymosomes play a key role during shown in Figure 5.
cellular communication that promotes sperm maturation
during spermatozoa passage into the microenvironment of the miRNAs and Spermatogenesis
epididymis (Belleannée, 2015). Spermatogenesis is a well-organized process that culminates
In general, miRNAs are post-transcriptional regulators in the production of sperm cells. Conditional knock-out
that play important roles in physiological processes and of the miRNA processing enzyme Dicer1 in Sertoli cells
their impairment results in different pathologies. These triggers male infertility due to the absence of sperm in
molecules have been shown to have great importance in the lumen of the seminiferous tubules and spermatogenesis
the regulation of spermatogenesis (Papaioannou et al., disorders (Papaioannou et al., 2009; Zimmermann et al.,
2009; Kotaja, 2014) and sperm maturation during sperm 2014), suggesting that the biogenesis of small non-coding
passage through the epididymis (Björkgren et al., 2012; RNA is important to spermatogenesis. Indeed, each cell
Jerczynski et al., 2016; Sipilä and Björkgren, 2016) as shown type of the seminiferous tubule has a different miRNA
in Figure 5. With the knowledge that miRNAs are very stable, profile that plays an important role in proliferation and
conserved bioactive molecules derived from the different differentiation (Hayashi et al., 2008; Smorag et al., 2012).
internal organs of the male reproductive tract, they could In pigs, miR-26a has been shown to inhibit proliferation
be considered potent fertility biomarkers. In light of these and to promote apoptosis in Sertoli cells, thus impairing
findings, sperm cells have gained the status of gametes that sperm production (Ran et al., 2018). In mice, miR−221 and
FIGURE 3 | Morpho-functional sperm features. Schematic figure representing the spermatozoa with satisfactory (left) and unsatisfactory (right) morpho-functional
features. Sperm acrosome membrane integrity, sperm plasma membrane integrity, sperm DNA integrity, low quantity of ROS, sperm mitochondrial membrane high
activity, high sperm motility and normal sperm morphology characterize the satisfactory morpho-functional sperm features. Sperm acrosome membrane damage,
sperm plasma membrane damage, sperm DNA fragmentation, high quantity of ROS, sperm mitochondrial membrane low activity, low sperm motility and abnormal
sperm morphology characterize the unsatisfactory morpho-functional sperm features.
miR−290 regulate the proliferation of spermatogonia and miRNAs and Sperm Maturation
primary spermatocytes (Smorag et al., 2012). In addition, Once produced in the testis, spermatozoa must pass through the
miR−203 modulates spermatocyte meiosis and the miR−34 epididymis to acquire their motility and oocyte-fertilizing ability.
family regulates the formation of spermatids (Smorag et al., During their passage through the different epididymal segments,
2012). In humans, pigs and mice, the miR−34 family is also spermatozoa come into contact with different repertoires of
important in the regulation of spermatogenesis (Bouhallier ncRNAs, transcripts and proteins that are released from the
et al., 2010; Zimmermann et al., 2014). Thus, miRNAs such epithelium of the epididymis mostly via epididymosomes
as miR−26a, miR−221, miR−290, miR−203, and the miR−34 (Sullivan et al., 2007; Belleannée et al., 2013; Sullivan, 2015;
have been proposed to contribute to spermatozoa production Reilly et al., 2016; Sharma et al., 2018). The transfer of proteins
(Figure 5), as their dysregulation results in disturbances and other contents from epididymosomes to spermatozoa has
in spermatogenesis and consequently a decrease in male been proposed to occur via the dynamin 1 mechanoenzyme
fertility potential. following the tethering of epididymosomes to specific sperm
FIGURE 4 | Intrinsic sperm features. Spermatozoon contributes early embryo development during mature oocyte fertilization, zygote formation and embryo cleavage
potentially with intrinsic sperm features such as: sperm proteins (e.g., PLCζ, to promote oocyte activation); sperm RNAs and microRNAs (miRNAs); sperm DNA, to
generate male pronucleus; sperm centrioles, to form sperm aster; and may contribute sperm mitochondria, promoting mtDNA heteroplasmy.
FIGURE 5 | Schematic figure demonstrating the contribution of microRNAs (miRNAs) in the reproductive events. The sperm-related-miRNAs molecules display
functions in spermatogenesis (testis), sperm maturation (epididymis caput, corpus, and cauda), sperm and seminal plasma interaction (e.g., epididymosomes) as
well as modulating early embryo development.
receptors located in the post-acrosomal region and the fusion of Schatten and Sun, 2009), as shown in Figure 4. Thus, centrioles
epididymosomes with the sperm membrane (Zhou et al., 2019). are organelles that ensure proper embryonic development.
In mice, sperm miRNAs present in the sperm are altered along the In addition, the proper formation of the sperm aster is an
epididymis (Figure 5), showing that the sperm miRNA content is important factor related to male fertility. For instance, large and
dynamic even following spermatogenesis (Nixon et al., 2015). well organized sperm aster formations in human spermatozoa
Conditional knock-out mice for the Dicer enzyme in principal correlate with samples that present with high fertility, while
cells of the epididymis dysregulated the differentiation process small and disorganized sperm aster formations correlate with
(Björkgren et al., 2012). Thus, biogenesis of ncRNAs seems low fertility samples (Navara et al., 1997). Therefore, beyond
to be important in the regulation of epididymal epithelium, the transmission of the centrioles by the sperm cells, the quality
sperm maturation and fertility. While miR-10a/b, −21a, −29c, of the sperm-borne centrioles is also important to embryo
−196b−5p, −199a, −200b/c, and −208b−3p accumulate development. It has recently been shown that sperm from
in spermatozoa during passage through the epididymis, non-rodent mammals are responsible for transmitting a second
miR−204b−5p and miR−375−3p are more abundant in atypical centriole to oocyte (Fishman et al., 2018). This centriole
spermatozoa from the caput and corpus of the epididymis (Reilly potentially forms the sperm aster and centrosome in the oocytes
et al., 2016; Sharma et al., 2018). Epididymosomes also display and, together with the first centriole, supports the fusion between
different miRNA profiles and traffic small RNAs to spermatozoa the female and male pronucleus.
(Sharma et al., 2018), including miR−143, −145, −199, and
−214 that are more abundant in epididymosomes present in the The Questioned Contribution of Sperm
caput of the bovine epididymis, and miR−395, −654, and −1224 Mitochondrial DNA to Embryo Paternal
that are more abundant in epididymosomes from the cauda
(Belleannée et al., 2013). In men with reproductive abnormalities,
Inheritance
miRNAs present in seminal plasma (miR−34c−5p, −122, Mitochondria are peculiar organelles that possess a specific
−146b−5p, −181a, −374b, −509−5p, and −513a−5p) DNA type: mtDNA. This organelle is essentially responsible
decreased in abundance in men presenting with azoospermia for energy production, which is important to the cell’s activity.
(absence of spermatozoa in the ejaculate) and increased During spermatogenesis, sperm mitochondria change from
in abundance in men presenting with asthenozoospermia a conventional to a condensed form that ensures more
(decreased sperm motility) (Wang et al., 2011). Although studies efficient energy production (Amaral et al., 2013). While some
have shown the importance of miRNAs along the epididymis, mitochondria are lost during spermatogenesis, between 22
further work is required to show how miRNAs are able to regulate and 75 mitochondria remain in the midpiece of the mature
sperm maturation during passage of spermatozoa through the spermatozoon depending on species (Figure 2) (Bahr and Engler,
epididymis and how regulation of motility acquisition and the 1970; Song and Lewis, 2008). Although spermatic mitochondria
ability of the sperm to become fertile occur. also contain mtDNA, the current paradigm is that the embryo
exclusively possesses maternal mitochondria and, consequently,
mitochondrial diseases are only inherited from the mother
(Hecht et al., 1984). However, the inheritance of paternal mtDNA
CONTRIBUTION OF SPERM INTRINSIC was suggested following the discovery of a high level of mtDNA
FACTORS TO POST-FERTILIZATION heteroplasmy, i.e., mix of maternal and paternal mitochondrial
PROCESSES genome, in mouse (Gyllensten et al., 1991) and human (17
individuals) (Luo et al., 2018) as schematized in Figure 4.
For many years it was postulated that sperm cells had the These findings open new research avenues to better understand
exclusive function of transporting genetic material to the oocyte. mitochondrial disease inheritance. However, further studies are
However, at the end of the 20th century, this sole contribution necessary to confirm this potential paradigm shift.
was challenged by the fact that specific sperm intrinsic factors,
including sperm centrioles, sperm proteins, ribonucleic acids and Contribution of Sperm Proteins to
mitochondrial DNA (mtDNA) could contribute to fertilization
Oocyte Activation
and/or early embryo development (Figure 6).
While mature spermatozoa are transcriptionally inactive upon
their entrance to the epididymis, their proteome is dynamic due
Contribution of the Sperm Centriole to to the acquisition of proteins from the surrounding fluid, the
Embryo Development cleavage of others from the sperm surface, and the rearrangement
Centrioles are cytoplasmic structures involved in cell division of acrosomal proteins during sperm capacitation. These proteins
and in the formation of cilia and flagella (Avidor-Reiss and ensure diverse functions at different stages including: sperm
Fishman, 2019). While spermatozoa present with centrioles maturation, e.g., the glioma pathogenesis-related 1-like protein
arranged in the neck of their tail (Figure 2), mature oocytes 1 (GLiPr1L1) that is acquired by spermatozoa and participates
lack centrioles (Simerly et al., 2018). Therefore, sperm-derived in the binding of sperm cells to zona pellucida (Caballero
centrioles ensure the formation of sperm aster and centrosome in et al., 2012); tagging of sperm sub-populations to ensure
the fertilized oocyte, which bring the female and male pronuclei heterogeneity, e.g., the sperm binding protein 1 (ELSPBP1) that
into close proximity and promote syngamy (Simerly et al., 1995; discriminates dead spermatozoa (D’Amours et al., 2012); and
FIGURE 6 | Timeline of the new findings regarding sperm morpho-functional and intrinsic features. The paternal DNA was considered as the sole intrinsic sperm
feature transferred from spermatozoon to oocyte until 1990s. In parallel, the sperm evaluation was limited to sperm conventional analyses (sperm motility and sperm
morphology/abnormalities). Proximal centrioles were then shown to be transmitted by sperm to the oocytes during fertilization for the first time in 1991
(Sathananthan et al., 1991). The sperm-borne PLCζ protein was shown as a promotor of oocyte activation in 2002 (Saunders et al., 2002). The delivery of RNAs
molecules was then revealed as transferred from sperm to the oocyte in 2004 (Ostermeier et al., 2004). In parallel, the sperm evaluation was updated to sperm
functional analyses (e.g., sperm plasma membrane integrity). In 2010s, small sperm-borne RNAs molecules (microRNAs/miRNAs) were shown as important to early
embryo development (Liu et al., 2012) potentially constituting a new group of sperm analyses composed by evaluation of molecular targets.
sperm ability to bind to the oocyte, e.g., Spermadhesin AWN potential contribution to the molecular regulation of embryonic
(AWN) that promotes sperm binding to oocyte’s zona pellucida development (Ostermeier et al., 2004). The discovery of
during fertilization (Kwon et al., 2014). zygote transcripts originating exclusively from spermatozoa
In addition to these proteins, sperm cells also acquire proteins opened new avenues into the investigation of sperm RNAs
during the final steps of spermatogenesis that are important to and their importance to embryo development (Ostermeier
oocyte activation, e.g., the sperm-specific WW domain-binding et al., 2005; Jodar et al., 2013). Most sperm RNAs are
protein (PAWP) and phospholipase C zeta (PLCζ) (Saunders transcribed and then stored or inactivated in spermatozoa
et al., 2002; Wu et al., 2007). Among these spermatic proteins, during spermiogenesis. During this stage of spermatogenesis,
PLCζ is an important player during fertilization, since it triggers histones are replaced by protamines in sperm DNA, changing
intracellular calcium impulses required for oocyte activation the shape of nucleosomes to a toroid structure with high sperm
(Saunders et al., 2002; Kashir et al., 2010). As a result of these nuclear condensation culminating in gene expression silencing.
impulses, and once fertilized, the metaphase II mature oocyte However, 1% of histones in mice and 10–15% of histones
is able to resume and progress through meiotic division. This in humans and cattle remain in the form of nucleosomes in
ensures the formation of female and male pronuclei, followed spermatozoa (Samans et al., 2014). The remaining nucleosomes
by syngamy (Figure 4). The presence of these proteins in in human and bovine spermatozoa are located in similar
spermatozoa positively correlates with male fertility (Saunders promoter regions, indicating transcriptional conservation of
et al., 2002; Aarabi et al., 2014). specific genes related to early embryonic development (Samans
et al., 2014). Even if the presence of RNAs is now accepted
Contribution of Sperm RNA Species to in spermatozoa, there is no evidence showing that RNAs
Embryo Development can be transcribed from DNA in mature spermatozoa. Most
Sperm RNAs of the RNAs present in sperm cells are thus potentially
The detection of sperm-borne RNAs, including clusterin acquired during spermatogenesis as well as during passage
and AKAP4, in embryos raised questions regarding their through the epididymis.
Even though sperm cells possess limited quantities of sperm i.e., the study of inheritance that is not based on DNA sequence,
RNAs, sperm-borne RNAs have been associated with many but on how the DNA sequence is utilized, miRNAs transmitted
relevant reproductive processes and several correlate with fertility by spermatozoa at the time of fertilization are considered
potential in the bull, including AK1 (adenylate kinase 1; epigenetic factors that have some effects on the offspring.
R2 = 0.90), IB (integrin β5, R2 = 0.95), NGF (R2 = 0.47), Environmental conditions such as toxicant exposure (Schuster
TIMP (tissue inhibitor of metalloproteinases, R2 = 0.39), SNRPN et al., 2016), mental stress (Rodgers et al., 2013), alcohol
(small nuclear ribonucleoprotein polypeptide N, R2 = 0.71) consumption (Rompala et al., 2018), and low protein or high fat
and PLCζ (phospholipase C, R2 = 0.69) (Kasimanickam et al., diet (Sharma et al., 2016) are considered principal modulators
2012). Similarly, the presence of 415 transcripts out of 24,000, of epigenetic markers by promoting alterations in the sperm
were found in different proportions between bovine spermatozoa “epigenome.” Rodgers et al. (2015) identified nine miRNAs that
presenting with high or low fertility potential (Feugang et al., had altered abundance in the sperm cells from mice submitted
2010). In human sperm cells, ANXA2 (Annexin A2), BRD2 to chronic stress. Surprisingly, the injection of these nine
(Bromodomain Containing 2), OAZ3 (Ornithine Decarboxylase miRNAs into embryos of non-stressed parents recapitulated the
Antizyme 3), PRM1 (Protamine 1) and PRM2 (Protamine 2) effects observed in parents undergoing chronic stress (Rodgers
transcripts were present in lower amounts in sperm cells with low et al., 2015). In a similar manner, regulation of another group
motility (Jodar et al., 2013). of ncRNAs in sperm cells, the tRNA-derived fragments, was
dependent on the diet of the sire and this influenced the offspring
Sperm miRNAs phenotype (Sharma et al., 2016). These studies show that sperm-
Until embryonic gene activation, i.e., when the embryo begins borne ncRNAs potentially own epigenetic influence. However,
to transcribe its newly formed genome, embryo development further studies are necessary in order to explain the mechanism
relies on the expression of maternal transcripts originating of action of these molecules in the offspring.
from the oocyte and, to a lesser extent, from spermatozoa.
MicroRNAs are also delivered by gametes to the embryos. Until
activation of the embryonic genome the abundance of miRNA LINKING MIRNAS TO MALE
transcripts present in the embryo decreases until embryonic gene (IN)FERTILITY STATUS
activation begins (Tang et al., 2007). Murine embryos produced
from Dicer enzyme knock-out oocytes fail to undergo the first Since miRNAs play an important role in the physiological
cell division, demonstrating the importance of the miRNAs or processes related to spermatogenesis, sperm maturation and early
other ncRNAs contributed by the female gamete during early embryonic development (Figure 5), these molecules have the
embryonic development (Tang et al., 2007). Similarly, murine potential to modulate sperm morpho-functional features as well
embryos generated with normal oocytes and Dicer or Drosha as male fertility ability. In that regards, different miRNA profiles
enzyme knock-out spermatozoa showed decreased development have been associated with different sperm quality levels and
rates, which recovered following the replacement of miRNAs fertility phenotypes from animal models and human clinical
(Yuan et al., 2016). In addition, the inhibition of miR-34c, which samples as listed in Table 2.
is exclusively delivered by sperm to the embryo in mice (Liu Functional studies have been carried out in knock-out
et al., 2012), but not in cattle (Tscherner et al., 2014), prevents mouse models to determine the contribution of individual
the first cleavage (Liu et al., 2012). Sperm-borne miR−449b miRNAs to male fertility (Comazzetto et al., 2014; Wu et al.,
overexpression in cloned embryos is associated with enhanced 2014). For instance, simultaneous deletion of miR34b/c and
cleavage rates at the 8-cell stage and lower rates of apoptosis in miR−449 triggered male infertility following the impairment
the blastocyst (Wang et al., 2017). Sperm-borne miR−216b was of spermatogenesis, motile ciliogenesis, and sperm maturation
recently shown to be present at higher abundance in spermatozoa (Comazzetto et al., 2014; Wu et al., 2014). While targeted
from low fertility bulls and is associated with a reduced first deletion is essential to unravel the function of specific miRNAs
cleavage rate as well as a reduced number of cells in blastocysts in vivo, phenotypic effects are rarely observed after invalidation
(Alves et al., 2019). In parallel, the RNAs that are acquired of a single miRNA-encoding gene due to compensatory
by spermatozoa during epididymal transit were shown to be functions of related miRNAs. These functional approaches were
essential for embryo implantation and development in mice complemented by several correlative studies performed in large
(Conine et al., 2018, 2019). Thus, despite the decrease in miRNA mammals and humans.
levels until activation of the embryonic genome, proper embryo Within livestock animals, bulls that presented satisfactory
development potentially relies on miRNAs delivered by the sperm quality and history of distinct fertility (high vs. low
gametes (Figure 5). fertility) displayed different profiles of sperm miRNAs that are
summarized in Table 2 (Fagerlind et al., 2015; Alves et al.,
Contribution of Small Non-coding RNAs 2019; Menezes et al., 2019). Similarly, spermatozoa from boar
to Transgenerational Inheritance of Traits featuring low motility and a high percentage of abnormalities,
The transgenerational inheritance of traits concerns the displayed a higher abundance of the miRNAs: let−7a, let−7d,
transmission of characteristics from the parents to the offspring, let−7e, and miR−22; and lower abundance of miR−15b (Curry
i.e., heritable phenotype, without altering the DNA sequence et al., 2011) compared with spermatozoa presenting with normal
(Weinhold, 2006). Following advances in the field of epigenetics, features. In addition, Capra et al. (2017) sorted spermatozoa
TABLE 2 | Sperm miRNAs associated with different infertility issues in porcine, human, bovine and mouse.
miRNAs Abundance Phenotype associate Specie References
let-7a Down-regulated Low abnormalities Porcine Curry et al., 2011

let-7a-5p Down-regulated High fertility Human Salas-Huetos et al., 2016
let-7d Down-regulated Low abnormalities/High motility Porcine Curry et al., 2011
let-7e Down-regulated Low abnormalities/High motility Porcine Curry et al., 2011
let-7f-5p Down-regulated High fertility Human Salas-Huetos et al., 2016
miR-9-3p Down-regulated High fertility Human Salas-Huetos et al., 2016
miR-15a Down-regulated High fertility Bovine Menezes et al., 2019
miR-15b Up-regulated Low abnormalities Porcine Curry et al., 2011
miR-15b Down-regulated Oligoasthenozoospermic Human Abu-Halima et al., 2013
miR-16 Down-regulated Oligoasthenozoospermic Human Abu-Halima et al., 2013
miR-17-5p Up-regulated High motility sperm Bovine Capra et al., 2017
miR-19a Down-regulated Oligoasthenozoospermic Human Abu-Halima et al., 2013
miR-19b-3p Down-regulated High fertility Bovine Fagerlind et al., 2015
miR-20a-5p Up-regulated High motility sperm Bovine Capra et al., 2017
miR-22 Down-regulated Low abnormalities Porcine Curry et al., 2011
miR-26a-5p Up-regulated High motility sperm Bovine Capra et al., 2017
miR-27a-5p Down-regulated High fertility Bovine Fagerlind et al., 2015
miR-29b Down-regulated High fertility Bovine Menezes et al., 2019
miR-30b-5p Down-regulated High fertility Human Salas-Huetos et al., 2016
miR-30d-3p Down-regulated High fertility Human Salas-Huetos et al., 2016
miR-33b Up-regulated High fertility Bovine Alves et al., 2019
miR-34a-5p Up-regulated High fertility Human Salas-Huetos et al., 2016
miR-34b Down-regulated Oligoasthenozoospermic Human Abu-Halima et al., 2013
miR-34c Up-regulated High fertility Mouse Liu et al., 2012
miR-34c-5p Down-regulated Oligoasthenozoospermic Human Abu-Halima et al., 2013
miR-34c-3p Down-regulated High fertility Bovine Fagerlind et al., 2015
miR-103a-3p Down-regulated High fertility Human Salas-Huetos et al., 2016
miR-122-5p Down-regulated High motility sperm Bovine Capra et al., 2017
miR-126-5p Up-regulated High fertility Bovine Alves et al., 2019
miR-127 Down-regulated Capacitated sperm Porcine Li et al., 2018
miR-148b-3p Down-regulated High fertility Bovine Fagerlind et al., 2015
miR-151-3p Up-regulated Capacitated sperm Porcine Li et al., 2018
miR-184 Down-regulated High motility sperm Bovine Capra et al., 2017
miR-205 Up-regulated High fertility Bovine Alves et al., 2019
miR-208a Up-regulated High fertility Human Salas-Huetos et al., 2016
miR-212-3p Up-regulated High fertility Human Salas-Huetos et al., 2016
miR-216b Down-regulated High fertility Bovine Alves et al., 2019
miR-320a Down-regulated High fertility Bovine Fagerlind et al., 2015
(Continued)
TABLE 2 | Continued
miRNAs Abundance Phenotype associate Specie References
miR-339a Down-regulated High fertility Bovine Alves et al., 2019

miR-346 Down-regulated High fertility Human Salas-Huetos et al., 2016
miR-449a Down-regulated Oligoasthenozoospermic Human Abu-Halima et al., 2013
miR-486-5p Down-regulated High motility sperm Bovine Capra et al., 2017
miR-487a Down-regulated High fertility Human Salas-Huetos et al., 2016
miR-502-5p Down-regulated High fertility Bovine Fagerlind et al., 2015
miR-518d-5p Down-regulated High fertility Human Salas-Huetos et al., 2016
miR-518f-3p Up-regulated High fertility Human Salas-Huetos et al., 2016
miR-520c-3p Down-regulated High fertility Human Salas-Huetos et al., 2016
miR-520d-3p Up-regulated High fertility Human Salas-Huetos et al., 2016
miR-542-5p Up-regulated High fertility Bovine Alves et al., 2019
miR-564 Up-regulated High fertility Human Salas-Huetos et al., 2016
miR-644a Down-regulated High fertility Human Salas-Huetos et al., 2016
miR-1249 Down-regulated High fertility Bovine Fagerlind et al., 2015
miR-1973 Up-regulated Asthenozoospermic Human Abu-Halima et al., 2013
according to their motility and showed that miR−17−5p, open avenues to the development of selection programs in
−20a−5p, −26a−5p, −122−5p, −184, and −486−5p were livestock animals. Targeting seminal miRNAs candidates as non-
differently displayed between the two subpopulations (Capra invasive biomarkers for male infertility could thus have profound
et al., 2017). Capacitated and non-capacitated spermatozoa economic impact on livestock industry.
also appeared to show different miRNAs profiles. For instance, In humans, sperm miRNA profiles from fertile and
Li et al. (2018) showed that miR−127, −151−3p, −152, infertile men accompanied or not by morpho-functional
−1285 were differently expressed in in natura (fresh) compared sperm alterations have been extensively investigated. In that
to capacitated boar sperm (Li et al., 2018). Based on that regards, men with reproductive disorders presented different
sperm miRNAs features, variation in expression of miRNAs sperm miRNA profiles compared with normospermic control
associated with economically important infertility issues might individuals: asthenozoospermia (low sperm motility) was
associated with higher abundance of 50 sperm miRNAs, which encompasses secretions from the different internal organs
among then hsa-miR−34b, −122, and −1973, and lower of the male reproductive tract. While both spermatic and extra-
abundance of 27 sperm miRNAs; oligoasthenozoospermia cellular RNA could be used as molecular targets for the non-
(low sperm concentration and low sperm motility) was invasive diagnosis of male infertility and reproductive system
associated with higher abundance of 42 miRNAs (hsa- diseases, a better understanding of miRNA-mediated regulation
miR−15b, −16, −19a, −34b, −34c−5p, −122, −449a e of reproductive functions is needed.
−1973 were the most abundant) and lower abundance of
44 miRNAs (Abu-Halima et al., 2013); teratozoospermia
(abnormal sperm morphology) was associated with the FINAL CONSIDERATIONS
downregulation of six miRNAs (hsa-miR−151−5p, −935,
−125a−3p, −132−5p, −320b, −195−5p) (Salas-Huetos et al., In summary, sperm cells should display specific factors to
2015). On the other hand, the sperm miRNA profile of men fulfill the “healthy sperm” concept, including proper morpho-
who presented normal semen quality (normozoospermic), functional features in addition to intrinsic factors. While sperm
but had a different fertility performance (fertile vs. infertile) cell morpho-functional attributes are not always sufficient to
showed that 57 miRNAs were presented at a different predict male fertility potential, spermatozoa carry different
abundance between the groups. Among these miRNAs miR- factors, including organelles (centrioles, mitochondria) and
15b−5p and 34a−5p, were related to pathways linked to molecules (proteins, RNAs, ncRNAs), which are involved in
embryonic development and chromatin remodeling (Salas- important steps of reproductive biology, e.g., spermatogenesis,
Huetos et al., 2016). Correlations were also observed between sperm maturation, fertilization and embryo development. These
sperm miRNA content and assisted reproductive technology factors constitute potential biomarkers of “healthy sperm” and
(ART) outcomes. For instance, the study from Cui et al. male fertility status and may become major assets for diagnosing
(2015) indicated that the proportion of good quality embryos instances of idiopathic male infertility in both humans and
obtained after 3 days post-ICSI positively correlated with the livestock animals. A better understanding of the mechanism
amount of hsa-miR−34c found in spermatozoa (Cui et al., of action of these sperm intrinsic factors in the regulation of
2015). While several confounding factors might be considered reproductive and developmental processes still presents a major
before drawing extrapolative conclusions, some spermatic challenge that must be addressed.
miRNAs may thus become predictive biomarker of defective
spermatozoa and ART outcome. Tentative explanations have
been proposed regarding the molecular mechanism involved AUTHOR CONTRIBUTIONS
in these correlative observations between sperm miRNA
profiles and fertility levels. For instance, hsa-miR−27a that CB, MA, and EC wrote the manuscript and designed the figures.
is increased in asthenoteratozoospermic patients regulates the All the authors revised the manuscript.
post-transcriptional expression of the Cysteine-Rich Secretory
Protein2 (CRISP2), which encodes for a protein playing a
role in sperm motility, acrosome reaction and gamete fusion FUNDING
(Zhou et al., 2017).
Overall, the different studies performed on human clinical We are grateful to the São Paulo Research Foundation (FAPESP)
samples as well as on animal models have shown that and the Natural Sciences and Engineering Research Council of
different profiles of sperm miRNA content can be observed Canada for financing the studies by the authors described in this
according to sperm morpho-functional features and fertility review (MA was funded by FAPESP Grants #2015/09154-6 and
status. Extracellular miRNAs correlated with male infertility #2016/23243-4. EC was funded by FAPESP Grant #2016/05395-1.
issues are also retrieved in seminal plasma (Barceló et al., 2018), CB was the recipient of an NSERC discovery grant).
REFERENCES C11. MHR Basic Sci. Reprod. Med. 13, 203–211. doi: 10.1093/molehr/
gal119
Aarabi, M., Balakier, H., Bashar, S., Moskovtsev, S. I., Sutovsky, P., Librach, C. L., Alves, M. B. R., de Arruda, R. P., De Bem, T. H. C., Florez-Rodriguez, S. A., Sá
et al. (2014). Sperm content of postacrosomal WW binding protein is related to Filho, M. F., de Belleannée, C., et al. (2019). Sperm-borne miR-216b modulates
fertilization outcomes in patients undergoing assisted reproductive technology. cell proliferation during early embryo development via K-RAS. Sci. Rep. 9, 1–14.
Fertil. Steril. 102, 440–447. doi: 10.1016/j.fertnstert.2014.05.003 doi: 10.1038/s41598-019-46775-8
Abu-Halima, M., Hammadeh, M., Schmitt, J., Leidinger, P., Keller, A., Meese, E., Amann, R. P., and Hammerstedt, R. H. (1993). In vitro evaluation of sperm quality:
et al. (2013). Altered microRNA expression profiles of human spermatozoa in an opinion. J. Androl. 14, 397–406. doi: 10.1002/j.1939-4640.1993.tb03247.x
patients with different spermatogenic impairments. Fertil. Steril. 99, 1249.e16– Amaral, A., Lourenço, B., Marques, M., and Ramalho-Santos, J. (2013).
1255.e16. doi: 10.1016/j.fertnstert.2012.11.054 Mitochondria functionality and sperm quality. Reproduction 146, 163–174. doi:
Agarwal, A., Mulgund, A., Hamada, A., and Chyatte, M. R. (2015). A unique 10.1530/REP-13-0178
view on male infertility around the globe. Reprod. Biol. Endocrinol. 13, 1–9. Ambros, V. (2004). The functions of animal microRNAs. Nature 431, 350–355.
doi: 10.1186/s12958-015-0032-1 doi: 10.1038/nature02871
Aitken, R. J., Wingate, J. K., De Iuliis, G. N., and McLaughlin, E. A. Avidor-Reiss, T., and Fishman, E. L. (2019). It takes two (centrioles) to tango.
(2007). Analysis of lipid peroxidation in human spermatozoa using BODIPY Reproduction 157, R33–R51. doi: 10.1530/REP-18-0350
Bahr, G. F., and Engler, W. F. (1970). Considerations of volume, mass, DNA, and Correa, J. R., Pace, M. M., and Zavos, P. M. (1997). Relationships among
arrangement of mitochondria in the midpiece of bull spermatozoa. Exp. Cell frozen-thawed sperm characteristics assessed via the routine semen analysis,
Res. 60, 338–340. doi: 10.1016/0014-4827(70)90526-4 sperm functional tests and fertility of bulls in an artificial insemination
Barceló, M., Mata, A., Bassas, L., and Larriba, S. (2018). Exosomal microRNAs program. Theriogenology 48, 721–731. doi: 10.1016/S0093-691X(97)00
in seminal plasma are markers of the origin of azoospermia and can predict 296-3
the presence of sperm in testicular tissue. Hum. Reprod. 33, 1087–1098. doi: Cui, L., Fang, L., Shi, B., Qiu, S., and Ye, Y. (2015). Spermatozoa micro ribonucleic
10.1093/humrep/dey072 acid–34c level is correlated with intracytoplasmic sperm injection outcomes.
Bartel, D. P. (2004). MicroRNAs: genomics, biogenesis, mechanism, and function. Fertil. Steril. 104, 312.e1–317.e1. doi: 10.1016/j.fertnstert.2015.05.003
Cell 116, 281–297. doi: 10.1016/S0092-8674(04)00045-5 Curry, E., Safranski, T. J., and Pratt, S. L. (2011). Differential expression of porcine
Bartel, D. P. (2018). Metazoan MicroRNAs. Cell 173, 20–51. doi: 10.1016/j.cell. sperm microRNAs and their association with sperm morphology and motility.
2018.03.006 Theriogenology 76, 1532–1539. doi: 10.1016/j.theriogenology.2011.06.025
Belleannée, C. (2015). Extracellular microRNAs from the epididymis as potential D’Amours, O., Frenette, G., Bordeleau, L.-J., Allard, N., Leclerc, P., Blondin, P.,
mediators of cell-to-cell communication. Asian J. Androl. 17, 730–736. et al. (2012). Epididymosomes transfer epididymal sperm binding protein 1
Belleannée, C., Calvo, É, Caballero, J., and Sullivan, R. (2013). Epididymosomes (ELSPBP1) to dead spermatozoa during epididymal transit in bovine. Biol.
convey different repertoires of microRNAs throughout the bovine epididymis. Reprod. 87, 1–11. doi: 10.1095/biolreprod.112.100990
Biol. Reprod. 89:30. doi: 10.1095/biolreprod.113.110486 Evenson, D. P., and Wixon, R. (2006). Clinical aspects of sperm DNA
Betlach, C., and Erickson, R. (1973). A unique RNA species from maturing mouse fragmentation detection and male infertility. Theriogenology 65, 979–991. doi:
spermatozoa. Nature 242, 114–115. doi: 10.1038/242114a0 10.1016/j.theriogenology.2005.09.011
Bianchi, E., Doe, B., Goulding, D., and Wright, G. J. (2014). Juno is the egg Izumo Fagerlind, M., Stalhammar, H., Olsson, B., and Klinga-Levan, K. (2015). Expression
receptor and is essential for mammalian fertilization. Nature 508, 483–487. of miRNAs in bull spermatozoa correlates with fertility rates. Reprod. Domest.
doi: 10.1038/nature13203 Anim. 50, 587–594. doi: 10.1111/rda.12531
Björkgren, I., Saastamoinen, L., Krutskikh, A., Huhtaniemi, I., Poutanen, M., Farrell, P. B., Presicce, G. A., Brockett, C. C., and Foote, R. H. (1998). Quantification
and Sipilä, P. (2012). Dicer1 ablation in the mouse epididymis causes of bull sperm characteristics measured by computer-assisted sperm analysis
dedifferentiation of the epithelium and imbalance in sex steroid signaling. PLoS (CASA) and the relationship to fertility. Theriogenology 49, 871–879. doi: 10.
One 7:e38457. doi: 10.1371/journal.pone.0038457 1016/S0093-691X(98)00036-3
Blom, E. (1973). The ultrastructure of some characteristic sperm defects and a Feugang, J. M., Rodriguez-Osorio, N., Kaya, A., Wang, H., Page, G., Ostermeier,
proposal for a new classification of the bull spermiogram. Nord. Vet. Med. 25, G. C., et al. (2010). Transcriptome analysis of bull spermatozoa: implications for
383–391. male fertility. Reprod. Biomed. Online 21, 312–324. doi: 10.1016/j.rbmo.2010.06.
Boerke, A., Dieleman, S. J., and Gadella, B. M. (2007). A possible role for sperm 022
RNA in early embryo development. Theriogenology 68(Suppl. 1), S147–S155. Fishman, E. L., Jo, K., Nguyen, Q. P. H., Kong, D., Royfman, R., Cekic, A. R.,
doi: 10.1016/j.theriogenology.2007.05.058 et al. (2018). A novel atypical sperm centriole is functional during human
Bouhallier, F., Allioli, N., Lavial, F., Chalmel, F., Perrard, M.-H., Durand, P., et al. fertilization. Nat. Commun. 9:2210. doi: 10.1038/s41467-018-04678-8
(2010). Role of miR-34c microRNA in the late steps of spermatogenesis. RNA Gadea, J., Parrington, J., Kashir, J., and Coward, K. (2013). “The male reproductive
16, 720–731. doi: 10.1261/rna.1963810 tract and spermatogenesis,” in Textbook of Clinical Embryology, eds K. Coward
Caballero, J., Frenette, G., Amours, O., Belleannée, C., Lacroix-Pepin, N., Robert, and D. Wells (Cambridge: Cambridge University Press), 18–26. doi: 10.1017/
C., et al. (2012). Bovine sperm raft membrane associated Glioma Pathogenesis- cbo9781139192736.005
Related 1-like protein 1 (GliPr1L1) is modified during the epididymal transit Gillan, L., Evans, G., and Maxwell, W. M. C. (2005). Flow cytometric evaluation of
and is potentially involved in sperm binding to the zona pellucida. J. Cell. sperm parameters in relation to fertility potential. Theriogenology 63, 445–457.
Physiol. 227, 3876–3886. doi: 10.1002/jcp.24099 doi: 10.1016/j.theriogenology.2004.09.024
Capra, E., Turri, F., Lazzari, B., Cremonesi, P., Gliozzi, T. M., Fojadelli, I., Gillan, L., Kroetsch, T., Chis Maxwell, W. M., and Evans, G. (2008). Assessment of
et al. (2017). Small RNA sequencing of cryopreserved semen from single bull in vitro sperm characteristics in relation to fertility in dairy bulls. Anim. Reprod.
revealed altered miRNAs and piRNAs expression between High- and Low- Sci. 103, 201–214. doi: 10.1016/j.anireprosci.2006.12.010
motile sperm populations. BMC Genomics 18:14. doi: 10.1186/s12864-016- Gyllensten, U., Wharton, D., Josefsson, A., and Wilson, A. C. (1991). Paternal
3394-7 inheritance of mitochondrial DNA in mice. Nature 352, 255–257. doi: 10.1038/
Celeghini, E. C. C., Alves, M. B. R., de Arruda, R. P., de Rezende, G. M., Florez- 352255a0
Rodriguez, S. A., and de Sá Filho, M. F. (2019). Efficiency of CellROX deep Hayashi, K., Chuva de Sousa Lopes, S. M., Kaneda, M., Tang, F., Hajkova, P., Lao,
red R and CellROX orange R fluorescent probes in identifying reactive oxygen K., et al. (2008). MicroRNA biogenesis is required for mouse primordial germ
species in sperm samples from high and low fertility bulls. Anim. Biotechnol. cell development and spermatogenesis. PLoS One 3:e0001738. doi: 10.1371/
[Epub ahead of print]. journal.pone.0001738
Celeghini, E. C. C., de Arruda, R. P., de Andrade, A. F. C., Nascimento, J., and Hecht, N. B., Liem, H., Kleene, K. C., Distel, R. J., and Ho, S. (1984). Maternal
Raphael, C. F. (2007). Practical techniques for bovine sperm simultaneous inheritance of the mouse mitochondrial genome is not mediated by a loss
fluorimetric assessment of plasma, acrosomal and mitochondrial membranes. or gross alteration of the paternal mitochondrial DNA or by methylation of
Reprod. Domest. Anim. 42, 479–488. doi: 10.1111/j.1439-0531.2006.00810.x the oocyte mitochondrial DNA. Dev. Biol. 102, 452–461. doi: 10.1016/0012-
Comazzetto, S., Di Giacomo, M., Rasmussen, K. D., Much, C., Azzi, C., Perlas, E., 1606(84)90210-0
et al. (2014). Oligoasthenoteratozoospermia and Infertility in Mice Deficient for Hermo, L., Oka, R., and Morales, C. R. (1994). Secretion and endocytosis in
miR-34b/c and miR-449 Loci. PLoS Genet. 10:e1004597. doi: 10.1371/journal. the male reproductive tract: a role in sperm maturation. Int. Rev. Cytol. 154,
pgen.1004597 106–189. doi: 10.1016/S0074-7696(08)62199-3
Conine, C. C., Sun, F., Song, L., Rivera-Pérez, J. A., and Rando, O. J. (2018). Small Hirohashi, N., and Yanagimachi, R. (2018). Sperm acrosome reaction: its site and
RNAs gained during epididymal transit of sperm are essential for embryonic role in fertilization. Biol. Reprod. 99, 127–133. doi: 10.1093/biolre/ioy045
development in mice. Dev. Cell 46, 470.e3–480.e3. doi: 10.1016/j.devcel.2018. Inaba, K. (2003). Molecular architecture of the sperm flagella: molecules for
06.024 motility and signaling. Zool. Sci. 20, 1043–1056. doi: 10.2108/zsj.20.1043
Conine, C. C., Sun, F., Song, L., Rivera-Pérez, J. A., and Rando, O. J. (2019). Irvine, D. S. (1998). Epidemiology and aetiology of male infertility. Hum. Reprod.
MicroRNAs absent in caput sperm are required for normal embryonic 13, 33–38. doi: 10.1093/humrep/13.1.33
development. Dev. Cell 50, 7–8. doi: 10.1016/j.devcel.2019.06.007 Jerczynski, O., Lacroix-Pépin, N., Boilard, E., Calvo, E., Bernet, A., Fortier, M. A.,
Cooper, T. G., and Yeung, C.-H. (2003). Acquisition of volume regulatory response et al. (2016). Role of Dicer1-dependent factors in the paracrine regulation of
of sperm upon maturation in the epididymis and the role of the cytoplasmic epididymal gene expression. PLoS One 11:e0163876. doi: 10.1371/journal.pone.
droplet. Microsc. Res. Tech. 61, 28–38. doi: 10.1002/jemt.10314 0163876
Jodar, M., Selvaraju, S., Sendler, E., Diamond, M. P., and Krawetz, S. A. (2013). The Piomboni, P., Focarelli, R., Stendardi, A., Ferramosca, A., and Zara, V. (2012). The
presence, role and clinical use of spermatozoal RNAs. Hum. Reprod. Update 19, role of mitochondria in energy production for human sperm motility. Int. J.
604–624. doi: 10.1093/humupd/dmt031 Androl. 35, 109–124. doi: 10.1111/j.1365-2605.2011.01218.x
Kashir, J., Heindryckx, B., Jones, C., De Sutter, P., Parrington, J., and Coward, K. Ran, M., Weng, B., Cao, R., Li, Z., Peng, F., Luo, H., et al. (2018). miR-26a
(2010). Oocyte activation, phospholipase C zeta and human infertility. Hum. inhibits proliferation and promotes apoptosis in porcine immature Sertoli cells
Reprod. Update 16, 690–703. doi: 10.1093/humupd/dmq018 by targeting the PAK2 gene. Reprod. Domest. Anim. 53, 1375–1385. doi: 10.
Kasimanickam, V., Kasimanickam, R., Arangasamy, A., Saberivand, A., Stevenson, 1111/rda.13254
J. S., and Kastelic, J. P. (2012). Association between mRNA abundance Raposo, G., and Stoorvogel, W. (2013). Extracellular vesicles: exosomes,
of functional sperm function proteins and fertility of Holstein bulls. microvesicles, and friends. J. Cell Biol. 200, 373–383. doi: 10.1083/jcb.
Theriogenology 78, 2007–2019. doi: 10.1016/j.theriogenology.2012.07.016 201211138
Kotaja, N. (2014). MicroRNAs and spermatogenesis. Fertil. Steril. 101, 1552–1562. Reilly, J. N., McLaughlin, E. A., Stanger, S. J., Anderson, A. L., Hutcheon, K.,
doi: 10.1016/j.fertnstert.2014.04.025 Church, K., et al. (2016). Characterisation of mouse epididymosomes reveals
Krawetz, S. A. (2005). Paternal contribution: new insights and future challenges. a complex profile of microRNAs and a potential mechanism for modification of
Nat. Rev. Genet. 6, 633–642. doi: 10.1038/nrg1654 the sperm epigenome. Sci. Rep. 6, 1–15. doi: 10.1038/srep31794
Kwon, W.-S., Rahman, M., Lee, J.-S., Kim, J., Yoon, S.-J., Park, Y.-J., et al. (2014). A Rodgers, A. B., Morgan, C. P., Bronson, S. L., Revello, S., and Bale, T. L. (2013).
comprehensive proteomic approach to identifying capacitation related proteins Paternal stress exposure alters sperm MicroRNA content and reprograms
in boar spermatozoa. BMC Genomics 15:897. doi: 10.1186/1471-2164-15-897 offspring HPA stress axis regulation. J. Neurosci. 33, 9003–9012. doi: 10.1523/
Kwon, W.-S., Rahman, M. S., Lee, J.-S., You, Y.-A., and Pang, M.-G. JNEUROSCI.0914-13.2013
(2015). Improving litter size by boar spermatozoa: application of combined Rodgers, A. B., Morgan, C. P., Leu, N. A., and Bale, T. L. (2015). Transgenerational
H33258/CTC staining in field trial with artificial insemination. Andrology 3, epigenetic programming via sperm microRNA recapitulates effects of paternal
552–557. doi: 10.1111/andr.12020 stress. Proc. Natl. Acad. Sci. U.S.A. 112, 13699–13704. doi: 10.1073/pnas.
Li, Y., Li, R. H., Ran, M. X., Zhang, Y., Liang, K., Ren, Y. N., et al. (2018). 1508347112
High throughput small RNA and transcriptome sequencing reveal capacitation- Rompala, G. R., Simons, A., Kihle, B., and Homanics, G. E. (2018). Paternal
related microRNAs and mRNA in boar sperm. BMC Genomics 19:1–12. doi: preconception chronic variable stress confers attenuated ethanol drinking
10.1186/s12864-018-5132-9 behavior selectively to male offspring in a pre-stress environment dependent
Liu, W., Pang, R. T. K., Chiu, P. C. N., Wong, B. P. C., Lao, K., and Lee, K. (2012). manner. Front. Behav. Neurosci. 12:257. doi: 10.3389/fnbeh.2018.00257
Sperm-borne microRNA-34c is required for the first cleavage division in mouse. Salas-Huetos, A., Blanco, J., Vidal, F., Godo, A., Grossmann, M., Pons, M. C.,
Proc. Natl. Acad. Sci. U.S.A. 109, 490–494. doi: 10.1073/pnas.1110368109 et al. (2015). Spermatozoa from patients with seminal alterations exhibit a
Luo, S., Valencia, C. A., Zhang, J., Lee, N.-C., Slone, J., Gui, B., et al. (2018). differential micro-ribonucleic acid profile. Fertil. Steril. 104, 591–601. doi: 10.
Biparental Inheritance of Mitochondrial DNA in Humans. Proc. Natl. Acad. Sci. 1016/j.fertnstert.2015.06.015
U.S.A. 115, 13039–13044. doi: 10.1073/pnas.1810946115 Salas-Huetos, A., Blanco, J., Vidal, F., Grossmann, M., Pons, M. C., Garrido, N.,
Mattick, J. S. (2001). Non-coding RNAs: the architects of eukaryotic complexity. et al. (2016). Sperm from normozoospermic fertile and infertile individuals
EMBO Rep. 2, 986–991. doi: 10.1093/embo-reports/kve230 convey a distinct miRNA cargo. Andrology 4, 1–9. doi: 10.1111/andr.12276
Menezes, E. S. B., Badial, P. R., El Debaky, H., Husna, A. U., Ugur, M. R., Kaya, Samans, B., Yang, Y., Krebs, S., Sarode, G. V., Blum, H., Reichenbach, M., et al.
A., et al. (2019). Sperm miR-15a and miR-29b are associated with bull fertility. (2014). Uniformity of nucleosome preservation pattern in mammalian sperm
Andrologia 52:e13412. doi: 10.1111/and.13412 and Its connection to repetitive DNA elements. Dev. Cell 30, 23–35. doi:
Morrell, J. M., Valeanu, A. S., Lundeheim, N., and Johannisson, A. (2018). Sperm 10.1016/j.devcel.2014.05.023
quality in frozen beef and dairy bull semen. Acta Vet. Scand. 60, 1–10. doi: Sathananthan, A. H., Kola, I., Osborne, J., Trounson, A., Ng, S. C., Bongso, A., et al.
10.1186/s13028-018-0396-2 (1991). Centrioles in the beginning of human development. Proc. Natl. Acad.
Navara, C. S., Hewitson, L. C., Simerly, C. R., Sutovsky, P., and Schatten, G. (1997). Sci. U.S.A. 88, 4806–4810. doi: 10.1073/pnas.88.11.4806
The implications of a paternally derived centrosome during human fertilization: Satouh, Y., Inoue, N., Ikawa, M., and Okabe, M. (2012). Visualization of the
consequences for reproduction and the treatment of male factor infertility. Am. moment of mouse sperm-egg fusion and dynamic localization of IZUMO1.
J. Reprod. Immunol. 37, 39–49. doi: 10.1111/j.1600-0897.1997.tb00191.x J. Cell Sci. 125, 4985–4990. doi: 10.1242/jcs.100867
Nixon, B., Stanger, S. J., Mihalas, B. P., Reilly, J. N., Anderson, A. L., Tyagi, S., Saunders, C. M., Larman, M. G., Parrington, J., Cox, L. J., Royse, J., Blayney, L. M.,
et al. (2015). The MicroRNA signature of mouse spermatozoa is substantially et al. (2002). PLC ζ: a sperm-specific trigger of Ca 2 + oscillations in eggs and
modified during epididymal maturation. Biol. Reprod. 93, 1–20. doi: 10.1095/ embryo development. Development 3544, 3533–3544.
biolreprod.115.132209 Schatten, H., and Sun, Q.-Y. (2009). The role of centrosomes in mammalian
Odia, M., Swanson, G., and Krawetz, S. A. (2018). A history of why fathers’ RNA fertilization and its significance for ICSI. Mol. Hum. Reprod. 15, 531–538.
matters. Biol. Reprod. 99, 147–159. doi: 10.1093/biolre/ioy007 doi: 10.1093/molehr/gap049
Oliveira, B. M., Arruda, R. P., Thomé, H. E., Maturana Filho, M., Oliveira, Schuster, A., Skinner, M. K., and Yan, W. (2016). Ancestral vinclozolin exposure
G., Guimarães, C., et al. (2014). Fertility and uterine hemodynamic in alters the epigenetic transgenerational inheritance of sperm small noncoding
cows after artificial insemination with semen assessed by fluorescent probes. RNAs. Environ. Epigenet. 2:dvw001. doi: 10.1093/eep/dvw001
Theriogenology 82, 767–772. doi: 10.1016/j.theriogenology.2014.06.007 Sellem, E., Broekhuijse, M. L. W. J., Chevrier, L., Camugli, S., Schmitt, E., Schibler,
Ostermeier, G. C., Goodrich, R. J., Diamond, M. P., Dix, D. J., and Krawetz, S. A. L., et al. (2015). Use of combinations of in vitro quality assessments to predict
(2005). Toward using stable spermatozoal RNAs for prognostic assessment of fertility of bovine semen. Theriogenology 84, 1447.e5–1454.e5. doi: 10.1016/j.
male factor fertility. Fertil. Steril. 83, 1687–1694. doi: 10.1016/j.fertnstert.2004. theriogenology.2015.07.035
12.046 Sharma, U., Conine, C. C., Shea, J. M., Boskovic, A., Derr, A. G., Bing, X. Y., et al.
Ostermeier, G. C., Miller, D., Huntriss, J. D., Diamond, M. P., and Krawetz, S. A. (2016). Biogenesis and function of tRNA fragments during sperm maturation
(2004). Reproductive biology: delivering spermatozoan RNA to the oocyte. and fertilization in mammals. Science 351, 391–396. doi: 10.1126/science.
Nature 429:2603. doi: 10.1038/nature02602 aad6780
Papaioannou, M. D., Pitetti, J. L., Ro, S., Park, C., Aubry, F., Schaad, O., et al. Sharma, U., Sun, F., Conine, C. C., Reichholf, B., Kukreja, S., Herzog, V. A.,
(2009). Sertoli cell Dicer is essential for spermatogenesis in mice. Dev. Biol. 326, et al. (2018). Small RNAs are trafficked from the epididymis to developing
250–259. doi: 10.1016/j.ydbio.2008.11.011 mammalian sperm. Dev. Cell 46, 481–494. doi: 10.1016/j.devcel.2018.06.023
Paul, J., and Duerksen, J. D. (1975). Chromatin-associated RNA content of Simerly, C., Manil-Ségalen, M., Castro, C., Hartnett, C., Kong, D., Verlhac, M. H.,
heterochromatin and euchromatin. Mol. Cell. Biochem. 9, 9–16. doi: 10.1007/ et al. (2018). Separation and loss of centrioles from primordidal germ cells
BF01731728 to mature oocytes in the mouse. Sci. Rep. 8, 1–17. doi: 10.1038/s41598-018-
31222-x
Simerly, C., Wu, G.-J., Zoran, S., Ord, T., Rawlins, R., Jones, J., et al. (1995). formation in C. elegans. Cell 75, 855–862. doi: 10.1016/0092-8674(93)
The paternal inheritance of the centrosome, the cell’s microtubule-organizing 90530-4
center, in humans, and the implications for infertility. Nat. Med. 1, 47–52. Wu, A. T. H., Sutovsky, P., Manandhar, G., Xu, W., Katayama, M., Day, B. N.,
doi: 10.1038/nm0195-47 et al. (2007). PAWP, a sperm-specific WW domain-binding protein, promotes
Sipilä, P., and Björkgren, I. (2016). Segment-specific regulation of epididymal gene meiotic resumption and pronuclear development during fertilization. J. Biol.
expression. Reproduction 152, R91–R99. doi: 10.1530/REP-15-0533 Chem. 282, 12164–12175. doi: 10.1074/jbc.M609132200
Smorag, L., Zheng, Y., Nolte, J., Zechner, U., Engel, W., and Pantakani, D. V. K. Wu, J., Bao, J., Kim, M., Yuan, S., Tang, C., Zheng, H., et al. (2014). Two miRNA
(2012). MicroRNA signature in various cell types of mouse spermatogenesis: clusters, miR-34b/c and miR-449, are essential for normal brain development,
evidence for stage-specifically expressed miRNA-221, -203 and -34b-5p motile ciliogenesis, and spermatogenesis. Proc. Natl. Acad. Sci. U.S.A. 111,
mediated spermatogenesis regulation. Biol. Cell 104, 677–692. doi: 10.1111/boc. E2851–E2857. doi: 10.1073/pnas.1407777111
201200014 Yuan, S., Schuster, A., Tang, C., Yu, T., Ortogero, N., Bao, J., et al. (2016).
Song, G. J., and Lewis, V. (2008). Mitochondrial DNA integrity and copy number in Sperm-borne miRNAs and endo-siRNAs are important for fertilization and
sperm from infertile men. Fertil. Steril. 90, 2238–2244. doi: 10.1016/j.fertnstert. preimplantation embryonic development. Development 143, 635–647. doi: 10.
2007.10.059 1242/dev.131755
Sullivan, R. (2015). Epididymosomes: a heterogeneous population of microvesicles Zegers-Hochschild, F., Adamson, G. D., Dyer, S., Racowsky, C., De Mouzon, J.,
with multiple functions in sperm maturation and storage. Asian J. Androl. 17, Sokol, R., et al. (2017). The international glossary on infertility and fertility care,
726–729. doi: 10.4103/1008-682X.155255 2017. Hum. Reprod. 32, 1786–1801. doi: 10.1093/humrep/dex234
Sullivan, R., Frenette, G., and Girouard, J. (2007). Epididymosomes are Zhou, J.-H., Zhou, Q.-Z., Yang, J.-K., Lyu, X.-M., Bian, J., Guo, W.-B., et al.
involved in the acquisition of new sperm proteins during epididymal (2017). MicroRNA-27a-mediated repression of cysteine-rich secretory protein
transit. Asian J. Androl. 9, 483–491. doi: 10.1111/j.1745-7262.2007.00 2 translation in asthenoteratozoospermic patients. Asian J. Androl. 19:591. doi:
281.x 10.4103/1008-682X.185001
Sullivan, R., and Saez, F. (2013). Epididymosomes, prostasomes, and liposomes: Zhou, W., Stanger, S. J., Anderson, A. L., Bernstein, I. R., De Iuliis, G. N.,
their roles in mammalian male reproductive physiology. Reproduction 146, McCluskey, A., et al. (2019). Mechanisms of tethering and cargo transfer during
R21–35. doi: 10.1530/REP-13-0058 epididymosome-sperm interactions. BMC Biol. 17:1–18. doi: 10.1186/s12915-
Tang, F., Kaneda, M., O’Carroll, D., Hajkova, P., Barton, S. C., Sun, Y. A., et al. 019-0653-5
(2007). Maternal microRNAs are essential for mouse zygotic development. Zimmermann, C., Romero, Y., Warnefors, M., Bilican, A., Borel, C., Smith, L. B.,
Genes Dev. 21, 644–648. doi: 10.1101/gad.418707 et al. (2014). Germ cell-specific targeting of DICER or DGCR8 reveals a novel
Tscherner, A., Gilchrist, G., Smith, N., Blondin, P., Gillis, D., and LaMarre, J. (2014). role for endo-siRNAs in the progression of mammalian spermatogenesis and
MicroRNA-34 family expression in bovine gametes and preimplantation male fertility. PLoS One 9:e107023. doi: 10.1371/journal.pone.0107023
embryos. Reprod. Biol. Endocrinol. 12:85. doi: 10.1186/1477-7827-12-85 Zini, A., and Agarwal, A. (2011). Sperm Chromatin. New York, NY: Springer.
Vincent, P., Underwood, S. L., Dolbec, C., Bouchard, N., Kroetsch, T., and Blondin, Ziyyat, A. (2001). Differential gene expression in pre-implantation embryos from
P. (2012). Bovine semen quality control in artificial insemination centers. Anim. mouse oocytes injected with round spermatids or spermatozoa. Hum. Reprod.
Reprod. 9, 153–165. 16, 1449–1456. doi: 10.1093/humrep/16.7.1449
Wang, C., Yang, C., Chen, X., Yao, B., Yang, C., Zhu, C., et al. (2011). Altered profile
of seminal plasma microRNAs in the molecular diagnosis of male infertility. Conflict of Interest: The authors declare that the research was conducted in the
Clin. Chem. 57, 1722–1731. doi: 10.1373/clinchem.2011.169714 absence of any commercial or financial relationships that could be construed as a
Wang, M., Gao, Y., Qu, P., Qing, S., Qiao, F., Zhang, Y., et al. (2017). Sperm- potential conflict of interest.
borne miR-449b influences cleavage, epigenetic reprogramming and apoptosis
of SCNT embryos in bovine. Sci. Rep. 7, 1–12. doi: 10.1038/s41598-017-1 Copyright © 2020 Alves, Celeghini and Belleannée. This is an open-access article
3899-8 distributed under the terms of the Creative Commons Attribution License (CC BY).
Weinhold, B. (2006). Epigenetics: the science of change. Environ. Health Perspect. The use, distribution or reproduction in other forums is permitted, provided the
114, 160–167. original author(s) and the copyright owner(s) are credited and that the original
Wightman, B., Ha, I., and Ruvkun, G. (1993). Posttranscriptional regulation publication in this journal is cited, in accordance with accepted academic practice. No
of the heterochronic gene lin- 14 by lin-4 mediates temporal pattern use, distribution or reproduction is permitted which does not comply with these terms.

Fcell 08 00791

Uploaded by

Copyright:

Available Formats

Fcell 08 00791

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Fcell 08 00791

Uploaded by

Copyright:

Available Formats

REVIEW

published: 21 August 2020

From Sperm Motility to Sperm-Borne

In addition to the paternal genome, spermatozoa carry several intrinsic factors,

Specialty section: INTRODUCTION

cells, and measurement of malondialdehyde concentration Intrinsic Sperm Features

result of their acquisition by sperm during passage through the

TABLE 1 | Correlation between sperm morpho-functional attributes and male fertility.

Sperm Definition Required equipment Correlation with fertility References

miRNAs Abundance Phenotype associate Specie References

let-7a Down-regulated Low abnormalities Porcine Curry et al., 2011

miRNAs Abundance Phenotype associate Specie References

miR-339a Down-regulated High fertility Bovine Alves et al., 2019

You might also like