Modes

of
repro-
duction
among
eukar-
yotes
OF WHAT BENEFIT IS SEX?
 Sexual periodicity is
much less strongly
developed in the male
than in the female.
 Among primates, the
male is sexually potent
throughout life.
 Rutting season- a brief
period of pronounced Public Display
sexual activity among of Affection
Not Allowed!
males; always coincides
with estrus or heat period of females.
 ESTRUS CYCLE- repeated series of
changes in the female reproductive
mechanisms.
1.Monestrous - breeds once a year
2.Polyestrous - several breeding periods in
a year:
 Pregnancy, starvation, extreme
exposure or sickness suppress estrus
 Domestication may cause shift to
polyestrous rhythm
 Critical initiating factors must be
present (ex. light in rabbits)
 GAMETOGENESIS
1.Origin of germ cells and
their migration to the gonads
2.Proliferation of germ cells
by mitosis
3.Meiosis
4.Final stages of maturation
for gametes
1. Origin of germ cells and their migration to the gonads
 The precursors of the gametes are the primordial
germ cells (PGCs). They form outside the gonads and
migrate into the gonads during development.
 In many species, a distinctive germ plasm exists. It
often contains the well conserved genes for the
Oskar, Vasa, Nanos, Tudor, and Piwi family proteins.
 In Drosophila, the germ plasm becomes localized in
the posterior of the embryo and forms pole cells, the
precursors of the gametes.
 In frogs, the germ plasm originates in the vegetal
portion of the oocyte.
 In birds, the germ plasm is first seen in the germinal
crescent. The germ cells migrate through the blood,
then leave the blood vessels and migrate into the
genital ridges.
 No germ plasm has been found in mammals;
PGCs arise from yolk sac endodermal cells with
the expression of the nuclear transcription factor
Oct 4 restrictively expressed in the inner cell
mass.
 During gastrulation, Oct 4 becomes expressed
solely in those posterior epiblast cells thought to
give rise to the PGCs.
 After that, this protein is seen only in the PGCs
and oocytes (Oct4 is not seen in the sperm after
the germ cells become committed to sperm
production.)
 Oct4 as an activator is sufficient to induce and
maintain pluripotency; if repressed, it induces
differentiation.
 Master
regulators of
differentiation:
Oct 4, Nanog,
Sox 2
External A+ and B-
signals activate
and repress
expression of
Oct4, Sox2 and
Nanog genes.
Oct4, Sox2 and
Nanog positively
regulate
pluripotency
genes and repress
differentiation
genes.
 (A). Final stages of migration of mammalian PGCs
through the hindgut into the 2 genital ridges which
will develop into gonads (B) The migrating PGCs in
an early mouse embryo are stained with a
monoclonal antibody (green). The remaining cells in
the embryo are stained with a lectin that binds to
sialic acid, found on the surface of all cells.
 In mammals, PGCs are determined to become
EGCs only after organogenesis has begun where
this transition is induced by somatic cells of the
gonad.
 The RNA-binding protein DAZL, is necessary in vivo
to restrict the developmental potential of the
germline.
 DAZL's absence leads to prolonged expression of
Nanog, which facilitates derivation of pluripotent
cell lines.
 DAZL mutations are associated with low sperm
count and premature ovarian failure.
 PGCs treated with FGF2 convert into embryonic
germ cells (EGCs) which have the potential to
differentiate into any cell type of the body, and
causes spontaneous gonadal teratomas.
A Proposed Model For Germ Cell Commitment In Mammals

Peter K. Nicholls et al. PNAS 2019;116:51:25677-25687

Cells deposited in extragonadal areas (mediastinum,
sacrococcygeal or oral regions) die, or develop into
teratomas / fetus-in-fetu which may contain mixtures
of differentiated tissues.
A teratoma complete with
teeth, eyes and veins; could Post-mortem
have a malignant potential. image of the
host twin,
demonstrating
significant
abdominal
distension
resulting from
the parasitic
twin and 3
intra-
peritoneal
teratomas.
 How Do the PGCs Know Where to Go?
 PGC migration in all species follows similar
steps: initiation of polarity and directed
migration, regulated migration by attractive
and repulsive cues, and termination of
migration at the site of gonad formation.
 In vitro evidence suggests that the genital
ridges of 10.5-day mouse embryos secrete a
diffusible TGF-β1-like protein that is capable of
attracting mouse PGCs.
 PGCs frequently use G protein-coupled
receptor signaling to reach their target
tissues, a mechanism found in many types of
migrating cells.
 CAMs play important roles in several steps of
PGC migration.
 Using alkaline phosphatase as a germ cell
"marker enzyme" for PGCs, they are shown to
migrate directionally to gonads by amoeboid
movement, presence of chemical attractants,
orientation by physical characteristics of the
fibronectin pathways, or may be provided by a
gradient of soluble protein.
 In mammals, a similar migration is seen, using
fibronectin pathways. Germ cells that lack the
integrin receptor for such ECM proteins
cannot migrate to the gonads.
PGCs migrate by extending filopodia (fine pseudopods) and they
can migrate between cells in tissues. They may follow ECM
components that are lined with fibronectin. PGCs (red) might use
their surface integrins to follow an embryonic "roadway" of ECM
to reach the genital ridges. The yellow is meant to reflect a
theoretical complex of ECM components (e.g., laminin,
fibronectin, collagen) rather than a single entity such as
fibronectin. Cells would stay within the yellow ECM region rather
than wander into adjacent regions because they would have
preferential adhesiveness ("stickiness") to the yellow "roadway".
  In mice, PGC
migration to the
genital ridges is
controlled by
the GPCR
CXCR4 and its
ligand SDF-1.
 SDF-1 is expressed by the somatic
cells of the genital ridge and PGCs
express CXCR4. Integrin β1 is also
required for this step. PGC motility
and survival requires the RTK c-Kit
and its ligand Steel.
 Steel is expressed by somatic cells surrounding
PGCs throughout migration.
 GENOMIC IMPRINTING
 Imprinting-switching off of genes that are not
expressed in development. Paternal or maternal
genomes maybe switched off during germ-cell
formation.
 Reciprocal imprinting: paternal genes promote
growth whereas maternal imprinting reduces
them (e.g. the paternal Igf-2 gene promotes a
large placenta; reduction of maternal gene
allows the mother to spread her resources over
all embryos)
 Imprinting is reversible; in mutants, failures and
abnormalities in development result from
mismatched timing of gene suppression.
 Mechanism of imprinting of the mouse Igf2 gene. On chromosomes
inherited from the female, a protein called CTCF binds to an insulator,
blocking communication between the enhancer (green) and the Igf2
gene (orange). Igf2 is therefore not expressed from the maternally
inherited chromosome. Because of imprinting, the insulator on the
male-derived chromosome is methylated; this inactivates the
insulator, by blocking the binding of the CTCF protein, and allows the
enhancer to activate transcription of the Igf2 gene.
 BIG DECISIONS:
Mitosis or Meiosis?
Sperm or Egg?
 In nematodes, the meiosis/mitosis decision is regulated by the
Delta proteins, which bind to the Notch proteins on the PGCs. On
the other hand, the signal for a germ cell to become either a sperm
or an egg is regulated at the level of translation of the fem-3 mRNA
containing a sequence that binds a repressor protein which is a
combination of the Nanos and Pumilio proteins.
 Retinoic Acid Determines The Timing Of Meiosis And Sexual
Differentiation Of Mammalian Germ Cells

(A) In female mouse embryos, RA secreted from the mesonephros reaches the gonad and triggers
meiotic initiation via the induction of Stra8 TF in female germ cells (beige). However, if activated Nanos2
genes are added to female germ cells, they suppress Stra8 expression, leading the germ cells into a
male pathway (gray). (B) In embryonic testes, Cyp26b1 blocks RA signaling, thereby preventing male
germ cells from initiating meiosis until embryonic day 13.5 (left panel). After embryonic day 13.5, when
Cyp26b1 expression is decreased, Nanos2 is expressed and prevents meiotic initiation by blocking
Stra8 expression. This induces male-type differentiation in the germ cells (right panel).
Meiosis
 Pre-meiotic Interphase

 Prior to meiosis, all chromosomes are duplicated in a process similar to

chromosome duplication prior to mitosis.
 Outside the nucleus of animal cells are two centrosomes, each
containing a pair of centrioles. The two centrosomes are produced by
the duplication of a single centrosome during pre-meiotic interphase.
The centrosomes serve as microtubule organizing centers (MTOCs).
Microtubules extend radially from centrosomes, forming an aster.
 Plant cells do not have centrosomes. Different kinds of microtubule
organizing centers serve as sites of spindle formation.
 Meiosis I: Prophase I

 At the start of prophase I, the chromosomes have already duplicated. During

prophase I, they coil and become shorter and thicker and visible under the light
microscope.
 The duplicated homologous chromosomes pair, and crossing-over occurs. Each
homologous chromosome pair is visible as a bivalent (tetrad), each consisting of two
sister chromatids. The sites of crossing-over are seen as crisscrossed non-sister
chromatids and are called chiasmata.
 The nucleolus disappears during prophase I.
 In the cytoplasm, the meiotic spindle, consisting of microtubules and other proteins,
forms between the two pairs of centrioles as they migrate to opposite poles of the cell.
 The nuclear envelope disappears at the end of prophase I, allowing the spindle to
enter the nucleus.
 Prophase I is the longest phase of meiosis, typically consuming 90% of the time for the
two divisions.
 Meiosis I: Metaphase I

 The centrioles are at opposite poles of the cell.

 The pairs of homologous chromosomes (bivalents), now as tightly
coiled and condensed as they will be in meiosis, become arranged
on a plane equidistant from the poles called the metaphase plate.
 Spindle fibers from one pole of the cell attach to one chromosome
of each pair (seen as sister chromatids), and spindle fibers from
the opposite pole attach to the homologous chromosome.
 Meiosis I: Anaphase I

 Anaphase I begins when the two chromosomes of each bivalent

(tetrad) separate and start moving toward opposite poles of the
cell as a result of the action of the spindle.
 In anaphase I the sister chromatids remain attached at their
centromeres and move together toward the poles.
 A key difference between mitosis and meiosis is that sister
chromatids remain joined after metaphase in meiosis I, whereas
in mitosis they separate.
 Meiosis I: Telophase I

 The homologous chromosome pairs complete their migration to the two

poles as a result of the action of the spindle. Now a haploid set of
chromosomes is at each pole, with each chromosome still having two
chromatids.
 A nuclear envelope reforms around each chromosome set, the spindle
disappears, and cytokinesis follows. In animal cells, cytokinesis involves
the formation of a cleavage furrow, resulting in the pinching of the cell
into two cells. After cytokinesis, each of the two progeny cells has a
nucleus with a haploid set of replicated chromosomes.
Meiosis I

Meiosis II:
Prophase II

 While chromosome duplication took place prior to meiosis I, no new

chromosome replication occurs before meiosis II.
 The centrioles duplicate. This occurs by separation of the two members
of the pair, and then the formation of a daughter centriole perpendicular
to each original centriole. The two pairs of centrioles separate into two
centrosomes.
 The nuclear envelope breaks down, and the spindle apparatus forms.
 Meiosis II: Metaphase II

 Each of the daughter cells completes the formation of a spindle

apparatus.
 Single chromosomes align on the metaphase plate. This is in
contrast to metaphase I, in which homologous pairs of
chromosomes align on the metaphase plate.
 For each chromosome, the kinetochores of the sister chromatids
face the opposite poles, and each is attached to a kinetochore
microtubule coming from that pole.
 Meiosis II: Anaphase II

The centromeres separate, and the two chromatids of

each chromosome move to opposite poles on the spindle.
Once sister chromatids separate, they are now called
daughter chromosomes in their own right.
 Meiosis II: Telophase II

 A nuclear envelope forms around each set of chromosomes.

 Cytokinesis takes place, producing four daughter cells (gametes, in
animals), each with a haploid set of chromosomes.
 Because of crossing-over, some chromosomes are seen to have
recombined segments of the original parental chromosomes.
Mammalian Synaptonemal Complex
Each axial element
(AE) is formed along
the chromosome
core of two sister
chromatids.
Heterodimers of
SYCP1 interdigitate
as transverse
filaments (TFs) in a
head-to-head
fashion. The SYCE1/
SYCE2/TEX12
The proteolytic destruction of the
complex stabilizes
synaptonemal complex unglues chromatid
TFs within the
arms, resolves the crossovers and allow
central element
homologues to separate at anaphase I. The
(CE). A series of
proteolytic destruction of the residual cohesin
chromatin loops are
complexes at the centromeres allows the
attached to the AEs.
sister chromatids to separate at anaphase II.
Independent
assortment,
and
crossing-
over leads to
Genetic
Variation