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Subject Topic Date Lecturer

OB-GYN Tranx No. 1 Gametogenesis 2009 December 7 Rex M Poblete, MD


Pages

Transcriber Ron Cortez

Batch 2012 / 2009-2010


OUTLINE
I. Gametogenesis A. Meiosis B. Spermatogenesis C. Oogenesis D. Vitellogenesis II. Development of the Ovum III. Fertilization
Sources: Powerpoint and Recordings

Subject Head

Ronnelaine Cortez

GAMETOGENESIS and DEVELOPMENT OF THE OVUM


I. Meiosis is the first step in gametogenesis: it is the separation of homologous chromosomes into haploid daughter cells.
A. Spermatogonia and oogonia are the germ cells that will eventually develop into the mature sperm or egg B. Primary spermatocyte or oocyte: the first step in this development is the duplication of homologous chromosomes to get ready for meiosis. C. Secondary spermatocyte or oocyte: the first meiotic division separates the homologous chromosomes from each parent D. Spermatids or eggs: the second meiotic division separates the 2 chromatids and creates 4 haploid cells In males, this eventually produces 4 sperm cells by the process of spermiogenesis. In females, it produces 1 egg and 3 polar bodies. This allows the egg to retain more cytoplasm to support early stages of development. Polar bodies are important for nourishment; they sustain the egg cell until it implants in the endometrium.

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II. Meiosis generates tremendous genetic diversity.


How many different types of gametes can be generated by an individual (male or female) with 23 different chromosomes? More than 223 or 8,000,000 different gametes.

III. The timing of meiosis differs in females and males.

Important to note: In males, the spermatogonia enter meiosis and produce sperm from puberty until death. The process of sperm production takes only a few weeks. Each ejaculation has 100 to 500 million sperm. However, females only produce egg cells from puberty until menopause. The mean age of menopause in Filipino women is around 50-52 years old. Perimenopause is the phase wherein females dont ovulate every month. Women aged 40 and above may miss their menses it means that they are not ovulating for that cycle. In females, this process is more complex. The first meiotic division starts before birth but fails to proceed. It is eventually completed about one month before ovulation. The second meiotic division occurs just before the actual process of fertilization occurs. When the egg is released from the ovary, thats the time when second meiotic division occurs and is completed before the union of the sperm and the egg. Thus, in females, the completion of meiosis can be delayed for over 50 years (because some women have late menopause and early puberty). This is not always good because some eggs may have chromosomal abnormality. This is why in females above 40, especially above 45, there is increased incidence of congenital anomaly, the more common one being Down syndrome. Advice pregnant patients above 45 to undergo screening for Down syndrome. Only 1 egg produced. In addition, all meiosis is ended in females at menopause.

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IV. Homologous chromosomes form the synaptonemal complex which facilitates crossing over and genetic diversity.
During meiosis, homologous chromosomes join together in pairs to form the synaptonemal complex. Each pair of chromatids is connected by axial proteins. The 2 homologous chromosomes are held together closely by central element proteins. A recombination nodule forms that contains enzymes for cutting and splicing DNA. Chromosomes are cut and joined crosswise at points called chiasmata, seen when they separate. The exchange of genetic material is evident when the chromosomes separate. This process is dangerous as it leads to deletions and duplications of genetic material. However, it is also valuable because it increases genetic diversity and facilitates evolution.

V. In older women, failure of the synaptonemal complex to separate properly can cause genetic disease.
Down syndrome is trisomy 21. It results in short stature, round face and mild to severe mental retardation. This is the failure of the 2 chromatids to separate during meiosis 2. It results in one oocyte receiving 2 instead of 1 chromatid. In older women, long term association of chromatids (i.e., over 50 years) results in the axial proteins failure to separate. Down syndrome occurs with a frequency of 0.2% only in women under 30 but rises at 3% in those over 45 years of age.

VI. Spermatogenesis occurs entirely in the seminiferous tubules.


The mammalian testes are divided into many lobules, and each lobule contains many tiny seminiferous tubules. Sperms develop in an ordered fashion in these tubules. Cells start to mature on the outside and move inward (towards the lumen) as they become mature sperm. Spermatogonia are the most primitive cells. They differentiate as primary spermatocyte secondary spermatid sperm are released into lumen. Sertoli cells are supporting cells that stretch from the lumen to the edge of the tubule. They nourish the developing sperm. They form a blood-testis barrier to control spermatogenesis (similar to the blood-brain barrier). These cells also inhibit spermatogenesis before puberty and stimulate the process after puberty.

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VII. Spermiogenesis is the maturation process into sperm.


A long process in which major structures are formed in order to make the sperm cell penetrate the egg cell and facilitate fertilization. The Golgi vesicles combine to form an acrosomal vesicle that lies over the nucleus. It is full of enzymes. Centrosomes start to organize microtubules into long flagella. Mitochondria start to localize next to the flagella to provide ready energy. The nucleus condenses in size and is stabilized by special proteins called protamines. The excess cytoplasm is pinched off as a residual body (no need for organelles and cytoplasmic proteins). Sperm cells are tiny, but are highly specialized for delivering the male genome: o Microfilaments shoot the acrosome into the egg to harpoon it and pull it in. o The acrosome has enzymes for breaking into the egg. o The midpiece has large numbers of mitochondria for energy. o The tail has a powerful flagellum for driving the sperm into the proximity of the egg (in humans, through the uterus and up into the oviduct).

VIII.

The goal of oogenesis is to produce one egg with massive amounts of cytoplasm.
In many organisms, the egg must contain all the nutrients to support the entire process of embryonic development. In humans, the egg does not need to grow so large because the fertilized egg only needs to support growth until the blastocyst implants in the uterus. The placenta then nourishes development. In some organisms such as frogs, oocytes grow to extremely large size and they have very active chromosomes that synthesize large amounts of RNA. In contrast to sperm which are tiny cells, oocytes are among the largest cells in the body. Oocytes contain Lampbrush chromosomes: Frog oocytes can contain 200,000 times as many ribosomes as a normal cell.

IX. Oocytes have a very small nucleus:cytoplasm ratio.


How long does it take for the blastocyst to travel from the fallopian tube into the endometrium, up to implantation? Around 7-9 days. This is why the oocytes have a small N:C ratio. It doesnt take that long for the placenta to take over nourishment of the blastocyst after the implantation. Most normal cells have several times as much cytoplasm as nucleus. This allows the nucleus to make enough mRNA and rRNA to keep up with the cytoplasm and cell needs. In some species, oocytes have a tremendously tiny nucleus to cytoplasm ratio. They must have a large amount of cytoplasm and ribosomes to make all of the proteins needed for embryonic development. The nucleus is just not large enough to keep up and maintain enough transcription to generate all of the needed components. However, oocytes have developed specializations to deal with this problem. o Ribosomal RNA genes are often amplified in oocytes. This allows more templates to transcribe more rRNA.

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X. Specializations allow the egg to accumulate cytoplasm: nurse cells allow oocytes of insects to produce massive amounts of RNA.

In Drosophila melanogaster, the oogonia are called cystoblasts, and they undergo an unusual specialization. They undergo multiple mitotic divisions, but fail to undergo cytokinesis (cell division). Thus, they all remain connected to the original cell as cystocytes. One of the lucky cystocysts becomes the oocyte. The other 15 become nurse cells for nourishment. They make large amounts of RNA and nutrients but they send it all to the oocyte. This allows the oocyte to accumulate massive amounts of cytoplasm to support development (15 nuclei instead of 1). What does a flys ovary look like?

XI. Vitellogenesis is the process of producing the major yolk proteins.


If you look at an ultrasound image of the embryo during the first trimester, you will see a yolk sac attached to the embryo. This yolk sac serves as nourishment for the fetus while the placenta is developing. Via ultrasound, if you see that the yolk sac is not growing or if the yolk sac is too big for the fetus, consider this as a bad sign because this means that the fetus is not feeding. The yolk sac must decrease in size as the placenta is developing. Yolk: animal eggs contain large amounts of protein, lipid, and glycogen to nourish the embryo. These materials are collectively called yolk. Yolk is minimal in animal eggs that sustain only the first portion of embryogenesis (humans and many mammals that have a placenta need only support cleavage for several days before implantation into the uterus). However, yolk is stored in large amounts in the eggs of birds and reptiles because their eggs have to support the entire process of development.
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Yolk proteins are synthesized in the liver in vertebrates, or in the fat body of insects (an analogous organ). Animal vegetal polarity: In eggs that have a lot of yolk, the yolk is concentrated in the vegetal pole. The animal pole contains the nucleus and relatively little yolk. The yolk in the vegetal pole interferes with cytokinesis during the process of cleavage leading to incomplete cleavage.

XII. Maturation processes prepare the oocyte for ovulation and fertilization.
Most oocytes of different species are arrested in the first meiotic division. o In human females, Meiosis I occurs prior to ovulation. Meiosis II occurs just before fertilization. Oocyte maturation begins officially when this block is removed and meiosis starts once again (meiosis II). 1. The nuclear membrane breaks down and DNA starts to condense into chromosomes. 2. The permeability of the oocyte plasma membrane changes so it can function outside of the ovary. 3. The plasma membrane develops receptors to interact with the sperm. As soon as a single sperm penetrates the egg, it makes the egg impermeable to other sperm cells. Fertilization occurs at different stages of oocyte maturation:

XIII. Control of oocyte maturation has been studied extensively in frogs.


Oocyte maturation is controlled by hormone interactions between the pituitary and follicle cells. Pituitary gonadotropin hormone stimulates follicle cells progesterone triggers oocyte maturation by activating c-mos expression. C-mos activates maturation-promoting factor, the same activity as M-phase promoting factor (MPF), that is composed of cyclin B and cyclin-dependent kinase 1. The exact mechanism isnt understood. If c-mos is inactivated by antisense oligonucleotides, no oocyte maturation occurs. On the other hand, if extra c-mos is injected it triggers oocyte maturation before it is ready. M-phase Promoting Factor (MPF) does many things, although the exact pathways have yet to be found. It causes breakdown of the nuclear envelope by phosphorylating nuclear lamins (proteins stabilizing the envelope), it triggers changes in the oocyte plasma membrane, it stimulates ovulation, and it causes condensation of chromosomes.
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XIV.

Development of mammalian oocytes occurs within the ovary.

In the mammalian ovary, the oocytes are closely associated with somatic cells called granulosa cells which aid oocyte maturation and ovulation. The timing of oocyte maturation and ovulation varies in different mammals. Ovulation can be stimulated by seasonal cues, the process of mating, or in primates, by the monthly cycle regulated by hormones such as estradiol, produced by the granulosa cells. In humans, it occurs monthly every 22-35 days.

XV. Eggs are protected by elaborate envelopes.


Vitelline envelope: a glycoprotein layer covers the plasma membrane of all eggs. This acts to protect the egg. Eggs that are deposited in water have a jelly-like coating that surrounds the egg (frogs eggs). Eggs that are deposited on land have particularly elaborate envelopes. The eggs of birds have a vitelline envelope, a fibrous layer, an outer layer of albumin (egg white), and a shell composed of calcium carbonate. The outer envelopes are synthesized in the oviduct after the egg has been fertilized.

How long should it take for fertilization to occur after ovulation? Within 24 hours after ovulation, fertilization should occur. If it occurs after 24 hours, pregnancy may still occur, but it may not be successful (spontaneous abortion). However, most women usually fail to notice spontaneous abortion because it may be mistaken as a normal menstrual period.

FERTILIZATION
Union of the spermatozoa and egg to produce a zygote. Occurs in the ampullary area of the fallopian tube. 16-cell morula blastocyst embryo What is the most common cause of spontaneous abortion in humans? Congenital anomaly (chromosomal abnormality) Females produce 1 egg cell per cycle. Males produce about 50-100 million per mL of ejaculate (after a 3-day abstinence). The most common cause of missed menses in a woman of reproductive age, provided that she is not pregnant, is the absence of ovulation. How long is human pregnancy on the average? 37-42 weeks (around 40 weeks) Pre-term Pregnancy: 20-36 6/7 weeks
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Abortion: Expulsion of the fetus before 20 weeks Post-mature Pregnancy: After 42 weeks Embryo: until after 8 weeks after fertilization or 10 weeks age of gestation The life span of a normal spermatozoon is roughly around 72 hours; meaning, if you had sexual intercourse at Day 12, there is a possibility of pregnancy. If you had sexual intercourse on Day 15 and she ovulated at Day 14, successful pregnancy is unlikely because the egg is most probably in the endometrial cavity already. Contraception: When is the safe time to have intercourse, granting that the female has a regular 28-day cycle? (Ovulation occurs at Day 14) = +/- 5 days . If the female is menstruating until Day 4 or 5, do it on Day 5-9 and Day 19 onwards. Live attenuated vaccines are not recommended for pregnant women. For example, MMR is recommended 3 months prior to planning conception. In general, after 30 weeks, most etiologic agents will not harm the fetus because the organ systems are already formed at that time. The critical period is during the first trimester.

PHASES OF FERTILIZATION

1. Passage of sperm through the corona radiata of the oocytes by the use of hyaluronidase which is released by the acrosome of the sperm and movement of the tail 2. Penetration of Zona Pellucidahelped by the enzymes esterase, acrosin, and neuraminidase. Once penetration occurs, there is a biochemical zona reaction that causes the egg to become impermeable to other sperm. 3. Fusion of the plasma membranes of the oocyte and sperm 4. Completion of second meiotic division of the oocyte and formation of the female pronucleus 5. Formation of the male pronucleus 6. Breakdown of pronuclear membranes, condensation of chromosomes, and arrangement of chromosomes for mitotic cell division

-END OF TRANSCRIPTIONHappy New Year, everyone! 1 OB-GYN Eval Coverage: Transcription No. 1 2 3 4
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January 11 (Monday), 3:00 PM Gametogenesis and Development of the Ovum Development and Physiology of the Decidua; Placenta and Placental Hormones Obstetrics in Broad Perspectives Fetal Morphological and Physiological Development

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