Introduction Quiz

Quiz 09

Spermatogenesis is initiated in the male testis with the beginning of Quiz

puberty. This comprises the entire development of the spermatogonia Quiz 10
(former primordial germ cells) up to sperm cells. The gonadal cords
that are solid up till then in the juvenile testis develop a lumen with the
start of puberty. They then gradually transform themselves into
spermatic canals that eventually reach a length of roughly 50-60 cm.
They are termed convoluted seminiferous tubules (Tubuli seminiferi
contorti) and are so numerous and thin that in an adult male testicle
their collective length can be 300 to 350 meters. They are coated by a
germinal epithelium that exhibits two differing cell populations: some
are sustentacular cells (= Sertoli's cells) and the great majority are the
germ cells in various stages of division and differentiation.

Fig. 8 - Convoluted seminiferous Fig. 9 - Convoluted seminiferous  Legend

tubules tubules
In certain
forms of
impotence and
for an
assisted
fertilization
the testicle
is
"biopsied":
from a piece
of testicular
parenchyma
the contorted
seminiferous
tubules are
extracted
with forceps
and unwound.
Fig. 8
Testicular
biopsy:
Pulling with
the forceps
makes the
testicular
canals
visible.
Fig. 9
The tubules
are clearly
visible using
 a binocular
magnifying
glass.
(Video, 390
kB)

Commentary
For an optimal sperm cell production a certain milieu is needed. By
transferring the testicles into the scrotum a testicular temperature 2-3
ºC lower than body temperature is attained. In addition, a slightly
elevated pressure from the surroundings is necessary. This is why when
the taut tunica albuginea is slit open, the testicular parenchyma bulges
out by itself. Evidently, both elevated pressure and lowered
temperature are necessary for producing sperm cells.

Fig. 10 - Convoluted seminiferous tubules  Legend

Fig. 10
Histological
preparation of
a section
through a
convoluted
seminiferous
tubule in an
adult. Outside
its basal
lamina a layer
of
myofibroblasts
and fibrocytes
surround the
tubule. The
germinal
epithelium lies
on the tubule
1 Basal lamina (membrane) (not recognizable) wall. One can
2 Myofibroblast recognize the
3 Fibrocyte spermatogonia
4 Sertoli's cell sitting on the
5 Spermatogonia basal lamina.
6 Various stages of the germ cells during The nuclei of
7 spermatogenesis the Sertoli's
8 Spermatozoon sustentacular
Lumen cells have a
rarified
chromatin and
the nuclei with
clear nucleolus
 that are often
oriented
perpendicular
to the basal
lamina. The
overall
picture,
though, is
dominated by
the cells
occupied with
spermatogenesis
.

The development of the germ cells begins with the spermatogonia at

the periphery of the seminal canal and advances towards the lumen over
spermatocytes I (primary spermatocytes), spermatocytes II
(secondary spermatocytes), spermatids and finally to mature sperm
cells.

Structure of the germinal epithelium Quiz

Quiz 08

The epithelium consists of Sertoli's sustentacular cells and the Quiz

spermatogenic cells. The Sertoli's cells form a single-layered lamina and Quiz 11
extend from the basal lamina to the tubule lumen. With their
labyrinthine cellular processes they surround the individual types of
germ cells more or less completely. Spermatogenesis is thus
accomplished in close contact with the Sertoli's cells, which not only
have supportive and nourishing functions, but also secrete hormones
and phagocytize cell fragments. Somewhat above the basal lamina they
are bound to each other through complicated occluding junctional
complexes (tight junctions), so that 2 separated compartments are
present in the epithelium: a basal one, in which the spermatogonia are
lined up, and a luminal one, in which all the other stages of
spermatogenesis are found.

Fig. 11 - Germinal epithelium  Legend

1 Peritubular cells
2 Basal membrane
3 Spermatogonia
4 Tight junction
5 Spermatocyte I
6 Spermatocyte II
7a Spermatids
7b Spermatids
8 Acrosome
9 Residual bodies
10 Spermatozoas
11 Cell nucleus of sustentacular cells (Sertoli)
A Basal zone
B Adluminal zone

Fig. 11
Schema of the
germinal
epithelium: The
supportive
(Sertoli) cells
sit on the
basal membrane.
Towards the
lumen of the
spermatogonia
(lowest row of
cells) the
Sertoli cells
are connected
 with each other
by the
occluding
junctional
complexes
(tight
junctions).
This seal gives
rise to the
blood-testicle
barrier. The
cytoplasm of
these
supportive
cells gets
formed into
complicated
processes
because they
surround all of
the cells
involved with
spermatogenesis
.

Commentary
Through the occluding junctional complexes of the Sertoli's cells a
"blood/testicle" barrier is created in the tubule. This means that outside
this barrier, in the tubular periphery, cells, substances and hormones
from the blood have unhindered access.
On the other hand, the inner compartment of the tubule is protected by
the barrier, which is selectively permeable and serves as an entry
check. This is of practical importance because haploid cells in the
inner part of the tubule exhibit surface antigenic properties, different
from all other body cells. They must thus be kept secluded from the
immune system of the organism by the "blood/testicle" barrier.

Developmental stages of spermatogenesis Quiz

Quiz 12

In the course of spermatogenesis the germ cells move towards the Quiz
lumen as they mature. The following developmental stages are thereby Quiz 13
passed through:

Quiz
 Quiz 14

 A-spermatogonium
 B-spermatogonium
 Primary spermatocyte (= spermatocyte order I)
 Secondary spermatocyte (= spermatocyte order II)
 Spermatid
 Sperm cell (= spermatozoon)

The spermatogenesis can be subdivided into two successive sections:

 The first comprises the cells from the spermatogonium up to and

including the secondary spermatocyte and is termed
spermatocytogenesis.
 The second one comprises the differentiation/maturation of the
sperm cell, starting with the spermatid phase and is termed
spermiogenesis (or spermiohistogenesis).

The temporal course of spermatogenesis

The approximate 64 day cycle of the spermatogenesis can be
subdivided into four phases that last differing lengths of time:

Mitosis of the 16 days Up to the primary spermatocytes

spermatogonia
First meiosis 24 days For the division of the primary
spermatocytes to form secondary
spermatocytes
Second meiosis A few For engendering the spermatids
hours
Spermiogenesis 24 days Up to the completed sperm cells
Total ~64    
days

Fig. 12 - The spermatogenesis generations  Legend

Fig. 12
The stem cell
population of
the germinal
cells lies on
the basal
lamina of the
convoluted
seminiferous
tubules.
These are Type
A
spermatogonia.
These cells
undergo
mitosis: one
of the
daughter cells
renew the
stock of type
A
spermatogonia,
the other
becomes a type
B
spermatogonia.
These divide
and their
daughter cells
migrate
towards the
lumen. In
roughly 64
days they
differentiate
themselves
thereby into
sperm cells up
to the outer
surface of the
epithelium
(one should
note that in
these cellular
divisions, the
separation of
the cytoplasm
is not
complete.
Whole networks
of connected
 cells arise.
So, for
example in the
last
generation,
the
spermatids,
far more cells
are bound to
each other
than as shown
here).

Spermatocytogenesis
Among the spermatogonia (all in all, over 1 billion in both testicles) Commentary
that form the basal layer of the germinal epithelium, several types In the
can be distinguished: certain type A cells are seen as spermatogonia heteronymou
that divide mitotically and reproduce themselves (homonymous s division
division), whereby the spermatogonia population is maintained. the
The beginning of spermatogenesis is introduced through the so- cytoplasmic
called heteronymous division, in which the daughter cells (second division is
group of type A cells) remain bound together by thin bridges of not
cytoplasm. Through the preservation of these cytoplasmic completed;
connections, spermatogonia are inducted into the spermatogenesis the
process. daughter
cells stay
After a further mitotic division type B spermatogonia are bound
engendered that also divide themselves mitotically into primary together
spermatocytes (I). through
thin
cytoplasmic
The freshly created primary spermatocytes (I) now enter into the bridges.
first meiosis. They then go immediately into the S phase (that is, into Also in the
the preleptotene meiosis), double their internal DNA, leave the basal subsequent
compartment and reach the special milieu of the luminal meiosis the
compartment. Following the S phase, these cells attain the complex cytoplasmic
stage of the prophase of the meiosis and become thereby noticeably division is
visible with a light microscope. incomplete,
so that
This prophase, which lasts 24 days, can be divided into five sections: from one
spermatogon
 Leptotene ium a
 Zygotene network of
 Pachytene daughter
 Diplotene cells
arises that
 Diakinesis
doubles in
size in
 each
generation.
The forming
of such
networks
assures
that all of
the
processes
in each
generation
occur in
step with
each other.
In the prophase in every germ cell a new combination of maternal and Commentary
paternal genetic material occurs. After the long prophase follow the about
metaphase, anaphase and telophase that take much less time. One meiosis
primary spermatocyte yields two secondary spermatocytes.

The secondary spermatocytes go directly into the second meiosis, out of which the
spermatids emerge. Since in the secondary spermatocytes neither DNA reduplication nor a
recombination of the genetic material occurs, the second meiosis can take place quickly. It
lasts only around five hours and for that reason secondary spermatocytes are rather seldom
seen in a histological section. Through the division of the chromatids of a secondary
spermatocyte, two haploid spermatids arise that contain only half the original DNA content.

Besides the sperm cells the spermatids are the smallest cells of the germinal epithelium. In a
process lasting several weeks (so-called spermiogenesis or spermiohistogenesis) they are
transformed into sperm cells with the active assistance of the Sertoli's cells.

Local course of spermatogenesis - the

spermatogenesis wave
In examining a cross-section of a Fig. 13 - Developmental stages  Legend
convoluted seminiferous tubule Fig. 13
one notices that cells appear in Various
groups having the same developmental
maturation stages. However, not stages in a
all the spermatogenesis stages are light
found in a cross-section. microscope
cross-
section
through a
convoluted
seminiferous
tubule.

1 Leptotene/zygotene of
the spermatocytes typ
2 I
Pachytene of the
3 spermatocytes
4 typ I
Young spermatids
5 Older spermatids
6 (sperm cells heads
can be recognized)
Sertoli's cells
Spermatogonia

On the one hand, the reason for this appearance lies in the fact that the
daughter cells, generated by each meiotic step, remain bound together
by thin cytoplasmic bridges. Thus with each meiotic step the following
generation is twice as large, until the cells have formed a relatively
complex network. The result is that cells of the same development
stages are seen there in groups. On the other hand, in addition, other
spermatogenesis generations are wound around each other in spirals
along the seminiferous tubule. This is why one meets with groupings of
various generations in a tubule cross-section. Thus, it is highly
improbable that all of the development stages will be seen in a single
section at the same time.

Fig. 14 - Spermatogenesis wave  Legend

 Fig. 14
This picture
shows three
waves of
spermatogenesi
s generations.

As in the diagram, spermatogenesis waves move in spirals - like a

corkscrew - towards the inner part of the lumen. Outside, on the edge of
the tubule and at the beginning of the spiral, lie the spermatogonia; and,
at the end of the spiral, the fully developed sperm cells are in the lumen.
From the diagram, it can be seen that several differing generations can
be found in a tubule cross-section. As time goes on, the wave of
spermatogenesis is shifted towards the right (as seen here) in order to
always newly begin again.

Spermiogenesis (spermatohistogenesis) and Quiz

structure of the sperm cell Quiz 15

The differentiation of the spermatids into sperm cells is called Quiz

spermiogenesis. It corresponds to the final part of spermatogenesis and Quiz 16
comprises the following individual processes that partially proceed at
the same time:
Quiz
 Nuclear condensation: thickening and reduction of the nuclear
size, condensation of the nuclear contents into the smallest Quiz 17
space.
 Acrosome formation: Forming a cap (acrosome) containing
enzymes that play an important role in the penetration through Quiz
the pellucid zone of the oocyte. Quiz 18
 Flagellum formation: generation of the sperm cell tail.
 Cytoplasma reduction: elimination of all unnecessary
cytoplasm.

Fig. 15 - Spermiogenesis  Legend

 Fig. 15
Three
differing
stages of
spermiogenesis
: on the left
a fresh
spermatid, on
the right an
immature sperm
cell, and in
the middle an
in-between
stage. A
rotation of
the nucleus
causes a
1 Axonemal structure, first flagellar primordium
repositioning
2 Golgi complex
of the
3 Acrosomal vesicle
acrosomal
4 Pair of centrioles (distal and proximal)
vesicle to
5 Mitochondrion
occur. This
6 Nucleus
inverts itself
7 Flagellar primordium
like a cap
8 Microtubules
over the
9 Sperm cells tail
nucleus that
10 Acrosomal cap
continues to
be condensed
(dotted line).
The cytoplasm
cell
components
that are no
longer needed
are discarded
and
phagocytized
by Sertoli's
cells. The
mitochondria
are packed
thickly
(tightly)
together
around the
beginning part
of the
flagellum
(mid-piece).
As a sign of
its
 immaturity,
the sperm cell
(on the right)
that has
issued into
the lumen
still has a
bit of
cytoplasm
around its
neck (compare
with fig. 16
below).

Nuclear condensation
The nucleus becomes smaller, denser and takes on a characteristic,
flattened form. Seen from above, the nucleus is oval and, from the
narrow side, is pear-shaped. The acrosome lies over the tip. Nucleus and
acrosome form the sperm cell's head that is bound to the mid-piece by a
short neck.

Acrosome formation
The Golgi complex engender the vesicles, which then merge into a
larger formation that settles close to the cell nucleus and finally inverts
itself like a cap over the largest part of the nucleus. The acrosome
corresponds functionally to a lysosome and thus contains lysosomal
enzymes (hyaluronidase among others).

Development of the flagellum

The future axonemal structure grows out of one centriole (distal). This
consists of a bundle of nine peripheral double microtubules and two
single ones in the center. During its development, through the rotation
of the nucleus and acrosomal vesicle, the flagellum primordium comes
to lie on the opposite side of the acrosome.

Four parts of the finished flagellum can be distinguished:

 The neck contains the two centrioles (proximal and distal)

among other things.
 The mid piece consists of a sheath of ring-shaped mitochondria
grouped around the axoneme to provide the energy for the
flagellar movement.
 The principle piece has a sheath of ring fibers around the
axoneme.
 The tail consists of only the 9+2 structure of the axoneme

The mature sperm cell is approximately 60 m long and completely

enveloped by the plasma membrane.
 Fig. 16 - The mature sperm cell  Legend
Fig. 16
The mature
sperm cell
is slender;
in the
middle part,
the
mitochondria
are thick
and ring-
shaped. The
DNA in the
nucleus is
maximally
condensed.
1 Plasma membrane
2 Outer acrosomal membrane
3 Acrosome
4 Inner acrosomal membrane
5 Nucleus
6 Proximal centriole
7 Rest of the distal centriole
8 Thick outer longitudinal fibers
9 Mitochondrion
10 Axoneme More info
11 Anulus More
12 Ring fibers information
about this
A Head illustratio
B Neck n
C Mid piece
D Principal piece
E Endpiece

Cytoplasmic reduction
The cytoplasm of the spermatids that is no longer needed is phagocytized by Sertoli's cells
or is disposed of in the lumen of the tubules. A clump of cytoplasm, though, can remain
hanging on the neck and mid piece of the sperm cell for a little while.

During sperm cell production considerable individual variations exist that are also
partially influenced by psychological factors. Per day roughly 100 million sperm cells are
produced. It is said that in each ejaculate an average number of 50-200 million sperm cells
are present (WHO standard value: over 40 million).

Leydig's interstitial cells and hormonal

regulation

Between the seminal canals lie Leydig's interstitial cells. These are
endocrine cells that mainly produce testosterone, the male sexual
hormone, and release it into the blood and into the neighboring tissues.
An initial active stage of these cells occurs during the embryonic
development of the testis. Later in juvenile life, due to the influence of
the LH (luteinizing hormone) secreted by the anterior hypophysis
(pituitary gland), Leydig's interstitial cells enter a second, long lasting
stage of activity. Together with the hormones secreted by the adrenal
cortex, testosterone initiates puberty and thus the maturation of the
sperm cells.

Fig. 17 - Leydig's interstitial cells  Legend

Fig. 17
Group of large
cells in the
interstice
between tubules.
Leydig's
interstitial
cells
characteristicall
y contain large
protein crystals
(crystalloids of
Reinke), the
importance of
which is unknown.
The crystals are
uncolored and
stand out as
light structures
1 Leydig's interstitial cells against the red
2 Crystalloids of Reinke cytoplasm of
Leydig's
interstitial
cells.

Commentary
Testosterone production is directed by LH (luteinizing hormone),
secreted by the anterior lobe of the hypophysis. Pronounced cycles in
hormone production, as are present in women, do not exist.
The second hormone secreted by the anterior hypophysis, FSH
(follicle-stimulating hormone) affects Sertoli's cells, in that it triggers
the formation of a testosterone-binding protein. Thereby testosterone
can be transported by Sertoli's cells into the luminal compartment and
there be concentrated. Testosterone is decisive for spermatogenesis.
Testosterone is also carried away via blood and lymph fluid.
Testosterone has effects on all tissues, especially also on the brain
during development as well as on the sexual organs.