Lecture 1: From Fertilization to Gastrulation Slide 1: Title slide: From Fertilization to Gastrulation

Main points: Covers Chapter 3 (First Week of Development: Ovulation to Implantation), Chapter 4 (Se on! Week of Development: "ilaminar #erm Dis ), an! Chapter $ (%hir! Week of Development: %rilaminar #erm Dis )

Slide 2: List of reasons for learning embryology

Main points: 1. This list provides students ith some of the facts about birth defects. 2. !n addition to those listed" birth defects are a ma#or contributor to our health care costs. $ust to get a ne born infant ith spina bifida home from the hospital can run up to %&''"'''" and then there are additional costs for the lifetime needs of the individual. &. There is almost no measure for the amount of human suffering birth defects can cause" both for the affected child and his or her family. !t is a very stressful situation for a family( divorce rates among those ith children ith birth defects are high. )nd if a couple thin*s they are not going to get pregnant" they should remember that +', of all pregnancies are unplanned.

Slide &: Graph sho

ing hen most structural birth defects are induced.

Main points: 1. This slide is one of those -ta*e.home message/ types. !t is crucial for everyone to understand that development happens fast and early in pregnancy even before omen realize they are pregnant. 2. 0uring the embryonic period 1&.2 ee*s( see slide3" the primordia of each organ system is established" and it is at this time that each organ is most sensitive to the induction of a birth defect. !t does not mean that the rest of gestation 1 ee* 4 to birth3 can be ignored from a birth defects standpoint" ho ever. For e5ample" the brain continues to develop even postnatally and remains sensitive to insult from many factors" including drugs li*e alcohol 1alcohol e5posure during gestation is the leading cause of mental retardation3" but the baby is less li*ely to have a ma#or structural defect induced after the eighth ee*. Therefore" since most omen do not go to their first prenatal visit until the eighth ee* 1i.e." after the period of greatest sensitivity3" it is imperative that birth defects prevention programs begin prior to conception! &. )nother issue is calculating embryonic age. 6n this slide and throughout the te5t" embryonic age is calculated from fertilization. 7ost physicians calculate the age as gestational or menstrual age and use the Last 8ormal 7enstrual 9eriod 1L8793 as the starting point. This practice is very inaccurate since only +', of omen ovulate ithin a :.day period around the midpoint of their cycle. Some actually ovulate ithin &.; days of the end of their last menstrual period. !n any case" since fertilization is supposed to occur at the midpoint of the cycle" the L879 method

Sadler, T.W.: Langmans Medical Embryology, 11e

Lippincott Williams & Wilkins, 2010

begins 2 ee*s prior to actual fertilization. Therefore" converting fertilization age to L879 age re<uires adding 2 ee*s.

Slide ;: =ross section of a uterus" uterine tube" and ovary

Main points: 1. Fertilization occurs in the ampullary region of the uterine 1Fallopian3 tube" and embryonic development begins immediately. 2. 0evelopment continues as the embryo moves do n the tube to ard the uterus. >y & days" the morula 1mulberry3 stage is attained( by ;.+ days" the blastocyst stage is observed" and the embryo enters the uterine cavity. &. )t +.? days" the blastocyst implants. Interestin& point: The process is not efficient. )ppro5imately +', of embryos are spontaneously aborted regardless of a oman@s nutritional" economic" or other status. 6f those embryos that are lost" +', ill have a serious chromosomal defect.

Slide +: 7orula stage and compaction

Main points: 1. A7orulaA means mulberry and is a stage attained at & days ith 2.1? cells. Bach cell is capable of forming a complete embryo 1totipotent3. 2. =ompaction begins hen cells on the outside seal themselves from the e5ternal environment by forming tight #unctions. Fluid is pumped to ard the inside and begins to displace the inner cells. )t this stage" outer cells begin to be committed to forming trophoblast 1placental3 cells" hile cells on the inside ill form cells for the embryo.

Slide ?: >lastocyst stage and implantation

Main points: 1. )s fluid accumulates in the center of the conceptus due to compaction and cells pumping the fluid in ard" a blastocyst is formed. This structure is li*e a tennis ball ith a li<uid center. =ells on the outside form the outer cell mass or trophoblast( hile cells on the inside at one pole form the inner cell mass or embryoblast. =ells from the embryoblast ill develop into the embryo and represent the source of embryonic stem cells used 1or not3 in research. They are nearly totipotent and can differentiate into virtually any type of cell or tissue given the right gro th factors and environment. This characteristic is hat ma*es them so valuable clinically. These are the cells that the Cnited States Government has declared off limits from funding by federal resources. 2. !mplantation begins ith attachment to the uterine epithelium. This process is affected by receptors on the epithelial cells that recognize specific proteins on the surface of the trophoblast cells. The process is similar to that involved in leu*ocytes stic*ing to the alls of blood vessels. &. )t appro5imately ? days" the blastocyst implants. !mplantation elicits an antibody response by the uterine epithelium because the embryo is a foreign body 1half its genes come from dad3. 8ormally" the embryo bloc*s this response" but in

Sadler, T.W.: Langmans Medical Embryology, 11e

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autoimmune diseases li*e systemic lupus erythematosus" the process may not function properly" possibly resulting in infertility. ;. The trophoblast is the invading tissue" and it differentiates into t o cell types: the syncytiotrophoblast" hich forms a syncytium and is actively invasive and secretes hormones li*e human chorionic gonadotropin 1h=G3" the basis for pregnancy tests( and the cytotrophoblast" hich provides a proliferative pool of cells that move into the syncytiotrophoblast. +. The invasion of the blastocyst is similar to gro th of a tumor and can sometimes become unregulated. The result is formation of a hydatidiform mole. 1See ne5t slide.3

Slide :: =haracteristics of hydatidiform moles

Main points: )s listed Interestin& point: Degulation of the gro th of moles is by the paternal genome. Thus" during the earliest stages of development" paternal genes regulate differentiation of the trophoblast 1placenta3" hile maternal genes regulate gro th of the embryoblast.

Slide 2: Depeat of the fertilization slide 1slide &3

Main point: 1. !mplantation occurs at the end of the first ee* and usually ta*es place along the anterior or posterior alls of the uterus. Sometimes" ho ever" it may occur at ectopic sites. 1See ne5t slide.3

Slide 4: Sites for ectopic pregnancies

Main points: 1. The most common site is in the uterine tube" but other sites may occur. 2. Bctopic pregnancies are due to the invasive nature of the trophoblast and abnormal transport or recognition of the blastocyst. &. They are dangerous because development may proceed for several ee*s or months before rupture of the site" including blood vessels" occurs" hich can result in severe hemorrhaging and maternal and fetal death.

Slide 1': B5ample of a tubal pregnancy

Main points: 1. This is the most common site for an ectopic pregnancy.

Slide 11: Bctopic pregnancy in the rectouterine pouch

Main points: 1. This is the most common site for an ectopic pregnancy in the peritoneal avit'. The embryo falls to this position because it is the lo est site in the cavity.

Slide 12: )ssisted reproductive technologies 1)DT3

Main points:

Sadler, T.W.: Langmans Medical Embryology, 11e

Lippincott Williams & Wilkins, 2010

1. The ne5t series of slides provides some information about assisted reproductive technologies 1in vitro fertilization3. They are relevant because 1, of all births in the Cnited States arise from these procedures" and there are significant complications ith these pregnancies.

Slide 1&: Graph sho

ing incidences and births from )DT procedures

Slide 1;: )dverse outcomes from )DT Slide 1+: Graph of adverse outcomes from )DT
Main points: 1. There is an increase in lo birth eight" very lo birth eight" and premature births among )DT babies. 7ost of this is due to the fact that the techni<ues often result in multiple birth pregnancies" hich are al ays high ris*.

Slide 1?: Graph sho

ing lo birth eight in )DT singletons

Main points: 1. Bven in singleton )DT pregnancies" there is an increased ris* for lo birth eight" possibly suggesting that something inherent in the procedures is having an adverse effect.

Slide 1:: )DT and birth defects

Main points: 1. There is an increase in birth defects among )DT pregnancies" yet most centers do not arn people about this possibility. 2. There are beginning to be some long.term follo .up studies of )DT infants in hich it appears that there may be some developmental issues that have not been detected at birth.

Slide 12: =ontinued differentiation of the trophoblast and embryo

Main points: 1. )bout 1 or 2 days after implantation" the embryo and trophoblast each begin to differentiate into t o layers: The embryoblast differentiates into a dorsal layer" the epiblast" and a ventral layer" the hypoblast( the trophoblast forms the syncytiotrophoblast and cytotrophoblast. 2. 9lacental development continues as the syncytiotrophoblast burro s deeper into the uterine tissue.

Slide 14: =ross sections through implantation sites. The slide is used for a demonstration of
both early placental and embryonic development. Main points (pla enta): 1. Trophoblast cells contribute to the fetal portion of the placenta. 2. 6n the ninth day" spaces 1lacunae3 form in the invading syncytiotrophoblast.

Sadler, T.W.: Langmans Medical Embryology, 11e

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&. Eithin a fe days" these spaces fill ith maternal blood as the syncytiotrophoblast erodes uterine vessels. ;. )s the spaces fill ith blood" finger.li*e pro#ections of syncytiotrophoblast covering a core of cytotrophoblast pro#ect into the blood.filled regions. These represent primary villi" and most form on the embryonic pole. +. )nother layer" the e5traembryonic mesoderm" forms around the embryo and yol* sac and inside the cytotrophoblast. !nitially" this layer is solid" but soon spaces appear causing a separation into t o layers: e5traembryonic splanchnopleuric mesoderm 1splanchnic or visceral layer3 around the yol* sac and embryo" and e5traembryonic somatopleuric mesoderm 1somatic or parietal layer3 lining the inside of the cytotrophoblast. 1)ll of these terms are potentially confusing. Therefore" it is perhaps best to refer to these layers as visceral and parietal because these are the terms that ill be used later for all other layers that relate to organ systems. For e5ample" the layer on the e5ternal surface of an organ is the visceral layer Fvisceral pleura" visceral peritoneum" etc.G" hereas the layer along the body cavity or all is called the parietal layer Fparietal pleura" parietal peritoneum" etc.G. Similarly" the mesoderm layer that lies ad#acent to the embryo and yol* sac is the visceral layer" hile the layer ad#acent to the trophoblast is the parietal layer3. The parietal layer eventually is referred to as the chorionic plate or chorion" and the space bet een the layers is called the chorionic cavity. >oth layers are continuous at the connecting stal* and ill be important for forming blood vessels.

Slide 14: Depeat to tal* about the embryoHs development

Main points (em(r'o): 1. 0uring the second ee*" the embryoblast forms t o layers: a dorsal epiblast and a ventral hypoblast. The structure is li*e an 6reo coo*ie ithout the icing. Bpiblast cells ill form all of the embryonic tissues and cells. The hypoblast ill be important for early signaling to establish the cranial.caudal a5is but ill eventually be displaced. 2. T o cavities are formed: the amnion and the yol* sac. )mnioblasts proliferate from the edges of the epiblast and secrete amniotic fluid. Iol* sac 1endoderm3 cells proliferate from the edges of the hypoblast to form the yol* sac. &. 8obody *no s hether or not the yol* sac is involved in embryonic nutrition" but later" it ill provide the first blood cells and the germ cells 1eggs and sperm3 for the embryo.

Slide 2': !llustration of sagittal section of the embryo and membranes that summarizes events
of the second ee* Main points: 1. )t the end of the second ee*" e are li*e a chic* embryo ith the embryonic disc 16reo coo*ie3 sitting on top of a yol* sac. 2. The second ee* can be called the A ee* of t o@s.A 1See slide.3 )lso" there are three cavities formed: amniotic" yol* sac" and chorionic.

Slide 21: 0orsal vie

of the embryonic disc 16reo coo*ie3 at the beginning of the third ee*

Sadler, T.W.: Langmans Medical Embryology, 11e

Lippincott Williams & Wilkins, 2010

Main points: 1. T o features appear in the epiblast: 13 the primitive strea*" a shallo groove at the caudal end of the embryo( and 23 the oropharyngeal membrane" a disc.shaped region of tightly adhering epiblast and hypoblast cells. This is the primitive membrane bet een the oral cavity and the pharyn5. !t ill eventually degenerate by programmed cell death to create an opening into the pharyngeal portion of the gut. 2. These t o features demonstrate that the head 1cranial3 and tail 1caudal3 ends of the embryo have been established.

Slide 22: !llustration sho

ing a midsagittal section through the embryonic disc

Main points: 1. Bstablishment of the cranial.caudal a5is re<uires molecular signals from t o regions. First" the anterior visceral endoderm 1)JB3" consisting of hypoblast cells at the cranial end of the embryonic disc" e5presses head.forming genes. These signals are reinforced by those from a region 1a shallo circular depression3 at the cranial end of the primitive strea* called the node. The main head.inducing gene from this region is goosecoid 1a transcription factor3. 2. 6ther signals from the node specify dorsal and ventral regions of the embryo by interacting ith bone morphogenetic protein 1>793. This protein is secreted as a gro th factor in all regions and cells at this stage of development. !f this protein is not inhibited" then mesoderm becomes -ventralized/ to form intermediate mesoderm 1urogenital system3 and lateral plate mesoderm 1limbs and ventral body all3. !f it is inhibited" then mesoderm becomes -dorsalized/ and forms para5ial mesoderm 1somites FvertebraeG3 and notochord. Genes e5pressed in the node inhibit >79s. Goosecoid upregulates chordin 1a gro th factor3" hile noggin" follistatin" and nodal 1all gro th factors3 are secreted by the node" and all of these proteins inhibit >79s.

Slide 2&: B5ample of con#oined t

ins Main points: 1. !n frogs" overe5pression of goosecoid causes double.headed tadpoles. 9erhaps one ay for this type of con#oined t ins to form is by the same mechanism" hereby too much signaling from goosecoid induces additional head regions.

Slide 2;: 0orsal vie

s of the embryonic disc at day 1+ hen laterality is established

Main points: 1. 8ot only does the early embryo *no its head from its tail" it also *no s its left from its right. 2. First fibroblast gro th factor 1FGF3 is secreted by cells in the node. This causes the gro th factor nodal to be secreted by the node and then concentrated to the left side by the action of cilia on cells in the node. The ciliated cells establish a polarized beat to the left side by some un*no n mechanism 1maybe electrical3. &. =oncentration of nodal on the left initiates a cascade of gene e5pression on that side" culminating in e5pression of the transcription factor 9!TK2( the master gene for establishing left.sidedness.

Sadler, T.W.: Langmans Medical Embryology, 11e

Lippincott Williams & Wilkins, 2010

;. 7echanisms to establish the right side are not clear but may involve e5pression of snail.

Slide 2+: )bnormalities resulting from problems establishing sidedness

Main points: 1. Situs inversus is a complete reversal of the organs and sidedness" hich is not usually a problem clinically. !ndividuals ith situs inversus typically do not have other abnormalities. 2. 2', of people ith situs inversus have Lartagener syndrome" hich includes a problem ith abnormal cilia function. Thus" their situs inversus probably relates to a cilia problem at the beginning of the third ee* hen cilia in the node should have s ept nodal to the left side. &. )s each organ is formed" it too e5presses the cascade of genes that regulates sidedness" for e5ample hen the heart loops or the liver or spleen is formed. ;. 9eople hose organs do not e5press this cascade properly have laterality se<uences in hich one or more organs are on the rong side but not all of them. 1They do not have complete situs inversus.3 !f the abnormality is primarily on the right" they have polysplenia 1the spleen is a right.sided organ3 or if the abnormality is primarily left. sided" they have asplenia. They also have a higher chance of having other congenital malformations 1unli*e people ith complete situs inversus3.

Slide 2?: 0orsal vie

s of the embryonic disc sho ing initial stages of gastrulation

Main points: 1. Gastrulation is the process of ma*ing the three germ layers from the epiblast by cell migration. !t transforms the embryonic disc from a bilaminar structure 16reo coo*ie ithout the icing3 to a trilaminar structure 16reo coo*ie ith the icing3. 2. Gastrulation begins hen cells of the epiblast proliferate and migrate to ard the primitive strea* 1arro s3.

Slide 2:: 0orsal vie

of the embryonic disc and a cross section through the primitive strea* sho ing the process of gastrulation Main points: 1. =ells of the epiblast move to ard the strea*" turn in ard" detach from their epiblast neighbors" and migrate beneath the remaining epiblast cells to establish t o ne cell layers. 2. Some of the first cells to migrate into the strea* displace the hypoblast cells" such that the hypoblast layer ill be completely replaced ith a ne set of cells. Thus" the hypoblast does not contribute any cells to the embryo@s development( all of them arise from the epiblast. &. )dditional cells assume an intermediate position bet een the ventral cells" replacing the hypoblast and cells that remain in the epiblast. This middle 1icing3 layer ma*es the embryo trilaminar.

Slide 22: 0orsal vie

of the embryonic disc sho ing direction of migrating cells and a sagittal section through the midline sho ing prenotochordal and notochordal cells

Sadler, T.W.: Langmans Medical Embryology, 11e

Lippincott Williams & Wilkins, 2010

Main points: 1. =ells move in through all regions of the primitive strea*" including the node. )s they move through the strea*" their fates are specified. For e5ample" cells moving through the most cranial aspect of the primitive node ill form prechordal and notochordal cells( #ust lateral to this region" cells ill form heart cell precursors and so on. 2. Some cells move through the cranial aspect of the node and migrate in the midline directly to ard the oropharyngeal membrane. These cells ill form prechordal and notochordal cells that ill be critical for inducing the brain and establishing the midline.

Slide 24: Sagittal 1)3 and cross sections 1> and =3 sho

ing notochord development Main points: 1. Sagittal section sho ing midline location of the prechordal and notochordal cells and the cut lines for > and =. 2. )s cells that ill form the notochord move through the node" they first intercalate themselves into the hypoblast to form the notochordal plate 1>3. &. )s differentiation continues" these cells detach from the hypoblast and form a continuous line of cells called the notochord. This structure is critical for establishing the midline and the a5is of the embryo. !t eventually forms the nucleus pulposus of each intervertebral disc.

Slide &': =ross section through the trilaminar embryonic disc sho

ing the & germ layers

Main points: 1. The result of gastrulation is formation of a trilaminar 16reo coo*ie complete ith icing3 embryo" and the three layers are called the germ layers because they ill -germinate/ all of the tissues and organs of the embryo. 2. The three layers include ectoderm" formed from cells that remain in the epiblast( mesoderm" formed from cells that migrate in through the primitive strea* to form the middle germ layer" including the prechordal plate and notochord( and endoderm" formed by cells migrating through the strea* and displacing the hypoblast cells. &. Gastrulation progresses in a cranial to caudal direction. Thus" the head region becomes trilaminar first" then the nec*" thora5" and so on do n the body a5is. The process starts at the beginning of the third ee* and continues until near the end of the fourth ee*. ;. Bctoderm basically represents tissues that come in contact ith the outside orld" such as the s*in" brain and spinal cord" and eyes and ears. +. 7esoderm forms the heart and urogenital system and the tissues that hold everything together 1most bones and connective tissue3. ?. Bndoderm is related to the gut and its derivatives.

Sadler, T.W.: Langmans Medical Embryology, 11e

Lippincott Williams & Wilkins, 2010