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Between week 1 and 6, female and male embryos are phenotypically indistinguishable, even though the genotype (XX or XY) of the embryo is established at fertilization. By week 12, some female and male characteristics of the external genitalia can be recognized. By week 20, phenotypic differentiation is complete. 2 The indifferent gonads develop in a longitudinal elevation or ridge of intermediate mesoderm called the urogenital ridge Primordial germ cells arise from the lining cells in the wall of the yolk sac at weeks 3-4. At week 4-6, primordial germ cells migrate into the indifferent gonad. Male germ cells will colonise the medullary region and the cortex region will atrophy. Female germ cells will colonise the cortex of the primordial gonad so the medullary cords do not develop. 3 4 5 Spermatogonia → spermatozoa. Spermatogenesis take about 70 day. Germ cells colonise the sex cords in the primordial gonad. These cords connect with the rete testis, the epididymis and the vas deferens. Before birth, the germ cells proliferate by mitosis to form spermatogonia stem cells. These begin mitosis to maintain a population of selfregenerating stem cells that remain available up to and beyond the age of 70, to allow for continuous sperm production at a high rate. 6 7 At puberty, the cords hollow out to form the seminiferous tubules, where sperms are produced. Each testis has 250-750 tubules, which empty into the rete testis, and from there form the epididymis. Spermatogonia (diploid 2n) (fixed number of mitotic divisions typically 64) → Primary Spermatocytes (diploid 4n) … all linked together by cytoplasm bridges. 8 Primary Spermatocytes (push their way towards the lumen of the tubule ) → meiosis (M) M1: two secondary spermatocytes (haploid 2n) M2: four Spermatids (haploid 1n) Each Spermatogonium yields up to 256 spermatids. Spermatids are re-modelled to form sperm by Spermiogenesis, and the cytoplasmic bridges between them are broken down before they are released into the tubule lumen to be washed down to the rete testis by fluid secreted from Sertoli cells 9 10 11 Spermiogenesis (1) formation of the acrosome, which covers half of the nuclear surface and contains enzymes to assist in penetration of the egg and its surrounding layers during fertilization (2) condensation of the nucleus; (3) formation of neck, middle piece, and tail; (4) shedding of most of the cytoplasm as residual bodies that are phagocytized by Sertoli cells. 12 13 Primordial germ cells colonise the cortex of the primordial gonad, becoming Oogonia. These proliferate rapidly by mitosis, so by 20 weeks of gestation there are over 7 million, however most of these die, leaving about 2 million that all begin meiosis before birth to become Primary Oocytes. The Oogonia’s entry into meiosis 1 is stimulated by Mesonephric cells. These are flattened epithelia cells, also called Follicular Cells or Granulosa Cells, which surround the primary oocytes to form Primordial Follicles. Meiosis is then arrested at the diplotene stage (a resting stage) of prophase due to Oocyte Maturation Inhibitor (OMI) secreted from the Follicular Cells. 14 A woman therefore has all the oocytes she will ever have at birth – no more can be formed later and all ova are produced from this ‘stock’, some of which may remain arrested for 50 years before further development. However, remaining in this arrested stage for many years increases the chance of cell damage, accounting for the increasing risk of foetal chromosomal abnormalities in pregnancies of older women. Beginning at puberty until the menopause about 40 years later, a small number of follicles (15-20) begin further development each month. Formation of a mature gamete requires the follicle to go through 3 stages. 15 › The primary oocyte grows dramatically, but does not re-start meiosis › Flat follicular cells become cuboidal Granulosa cells › Granulosa cells secrete glycoprotein to surround the oocyte with a Zona Pellucida › Surrounding connective tissue (stroma) cells form a Theca Folliculi Inner Theca Interna that is vascular and endocrine Outer Theca Externa that is a fibrous capsule › Theca and Granulosa cells collaborate to secrete oestrogens 16 17 › Granulosa cells continue to proliferate and a fluid appears between them, forming the antrum › As more fluid forms, this secondary or Graafian follicle expands › Expands to 2mm diameter without stimulation from reproductive hormones › Continued development depends on reproductive hormones. FSH – Binds only to Granulosa cells LH – Binds only to Thecal cells › Under the influence of LH, Thecal cells secrete androgens, which are converted to oestrogens by the Granulosa cells under the influence of FSH 18 19 › Begins 37 hours before ovulation › Oestrogen causes receptors for LH to appear on outer Granulosa cells › LH surge stimulates these receptors, leading to rapid changes in the follicle › Within 3 hours of the LH surge, the follicle restarts meiosis, and the first › › › › › › meiotic division is completed. This division is asymmetric; cytoplasm remains with one daughter cell and the other forms a condensed polar body. The secondary follicle then enters meiosis II and arrests again 3 hours prior to ovulation. Follicle size increases dramatically by increase in antral fluid volume to 25mm diameter Structure begins to weaken LH stimulates collagenase activity leading to follicle rupture Ovum is carried out in the fluid and gathered up into the fallopian tube by fimbria Meiosis is not completed unless the ovum is fertilised 20 21 22 Pre-ovulatory follicle: Secondary follicle mature + LH surge → Pre-ovulatory growth phase: 1. MI completed → unequal size daughter cells each with 23 chromosomes: secondary perivetelline space ). 2. oocyte and 1st polar body (in Secondary oocyte begin MII but arrested in metaphase about 3 hours before ovulation where MII completed only if oocyte fertilized. If oocyte not fertilized → degenerate after about 24 hours. 1st PB also undergo 2nd division (MII). 23 The remains of the follicle (granulosa cells and theca interna) re-organise themselves into a corpus luteum cells, which secretes progesterone and oestrogen under the influence of LH. o In humans the corpus luteum lives for 14 days before regressing spontaneously, in the absence of a fertilization, where it called corpus albicans. o If the oocyte is fertilized, degeneration of the corpus luteum is prevented by human chorionic gonadotropin (hCG), a hormone secreted by the syncytiotrophoblast of the developing embryo. o The corpus luteum continues to grow and forms the corpus luteum of pregnancy (corpus luteum graviditatis). o 24 By the end of the third month, this structure may be 1/3 to 1/2 of the total size of the ovary. Luteal cells continue to secrete progesterone until the end of the fourth month; thereafter, they regress slowly as secretion of progesterone by the trophoblastic component of the placenta becomes adequate for maintenance of pregnancy. Removal of the corpus luteum of pregnancy before the fourth month usually leads to abortion. 25 Conception: it is formation of a zygote resulting from the union of a sperm and egg cell (oocyte) by the process of fertilisation that take place in ampullary region of fallopian tube. Sperm transport through the cervix and uterus Immediately after ejaculation in the vagina, the semen first coagulates due to the action of clotting factors (fibrinogen). This is to prevent sperm being physically lost from the vagina. 10 – 20 minutes later the semen re-liquefies by the action of enzymes found in prostatic secretions. The vast majority of sperm do not enter the cervix of the uterus and are lost by leakage from the vagina. Those that do enter the uterus have to travel 15 – 20cmto reach the uterine tube, a journey that may last a few hours. Transport of sperm is as a result of their own propulsive capacity and the fluid currents caused by the action of ciliated cells in the uterine tract. Capacitation of sperm and the acrosome reaction During their passage through the uterus to the uterine tube, sperm undergo a series of maturational changes, Capacitation and the Acrosomal Reaction. Both capacitation and the acrosomal reaction are induced by an influx of calcium and a rise in cAMP in spermatozoa. Capacitation Further maturation of sperm in female reproductive tract (6 – 8 hours) Sperm cell membrane changes to allow fusion with oocyte cell surface › Removal of glycoprotein coat Tail movement changes › Beat Whip-like action Sperm become responsive to signals from the oocyte Acrosomal Reaction Capacitated sperm comes into contact with the oocyte zonapellucida Membranes fuse Start of reaction Acrosome swells and liberates its contents by exocytosis Proteolytic enzymes and further binding facilitate penetration of the zonapellucida by the sperm (takes about 15 minutes) Mechanisms involved in fertilisation of the ovum By the time of ovulation, the ovum (primary oocyte) in the ovulatory follicle has completed its first meiotic division to form a secondary oocyte. Secondary Oocyte › Haploid number of chromosomes and bulk of cytoplasm First Polar Body › Remaining haploid number of chromosomes The secondary oocyte, surrounded by follicular cells (cumulus) embedded in a gelatinous matrix, is released from an ovulatory follicle and picked up by the fimbria of the uterine tube and guided into its lumen by the ciliary movements of epithelial cells towards the ampulla, the site of fertilisation, where the oocyte and sperms come together. Only one sperm penetrates the cytoplasm of the ovum and its nucleus fuses with the nucleus of the ovum. This forms the zygote. Sperm entry to oocyte will cause: 1.Cortical and zona reaction so oocyte become imper meable to further sperm preventing polyspermia. 2. Resumption of second meiotic division. 3. Metabolic activation of oocyte. So the main results of fertilization are: 1.Restoration of the diploid number of chromosome. 2.Determination of sex of new individual. 3. Intiation of cleavage. Within a few hours the zygote begins to divide by a series of mitotic cell divisions known as cleavage to form a ball of cells called the morula and then a hollow structure, the blastocyst. During this transformation process it is gradually transported along the uterine tube towards the uterus. By the time the blastocyst enters the uterine cavity (4 – 5 days after fertilisation), the endometrium is ready to receive it for pregnancy to be established. After a day or so in the uterine cavity the blastocyst attaches itself to the uterine endometrium – implantation.