9-Role of the Organizer
• At the end of gastrulation and neuration have an embryo with the three germa layers and axis • Everything is in respect to dorsal mesoderm • Mesoderm frms with dorsal zis on top • Mesoderm s induced • Activin is an important player; not the most importnant player but they work • Activin is a morphogen • Gooecoid defines dorsal organizer • Different mesoderms is due to different concentrations of moprogogen
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• When gastrulation starts, has tissue above dorsal lip • Organizer is comitted, gives rise to dorsal mesoderm • If you put the organizer elsewhere get 2 dorsals • Dorsal organizer is set up opposite sperm entry point • First cleavage plane defines left and right, goes through the organizer • Grey crescent develops; tells where dorsal is because of crodical rotation • If divide embryo and don’t give it grey crescent, develops abnorally • If ectoderm is transplanted before it is indeced, it will remian ectoderm • If wait unitl gastrulation prgresses; ectoderme is being induced and changed(knows position is neural). If transplant to another embryo will form neural tissues
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• Red band forms mesoderm • If take ac and vp, and put them together, will stell get signalling and form mesoderm (wont get any patterning) • PCR for mesodermal genes. If take late state change embryo that has a heart, will have the gene (cardiac actin). Wont have any in ac, wont have them in vp. However if u put ac and vp together, may get cadiac actin (induced the right type of actin)
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• Organizer is responsible for 1o embryonic inductuion • 4 function: 1. Make something called the prechordal plateà self determing, forms this all by itself 2. Starts off as ring of mesoderm. Notochord (dorsal mesoderm). The closer u are to them, the more dorsal character you are. Its whats maing somites dorsal. 3. Makes ectoderm near it neural, because of signalling 4. Intiates gastrulation
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• Cortucal rotationà rotates respect to inside of egg • Does so because of mt lining cortex, outside of egg • Before fert, mt will not be organized, after fert mt much better organized, soerm bring centriole • Centriole of mt organized centrer; beofre centrioles play role in mt apparatus. Centriole is mtoc allows mt to polmerate away from sperm that just came in • Provides track • When it rotates, cytoplasm floats around. When it swirls, able to see the grey crescent (dorsal side). First cleavage goes theough the grey crescent
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• Dsh is on vegetal side in cortex • Sperm ferts in animal half. When fert brings in centriole. When centriole comes in; polymerizes green tract. Causes rotation of the cortex (causes things insde to swil around), also gives dsh a tract to move. Kinesin moves dsh to move. • Polymerization downwards in respect to gravity. If gravity isn available polymerizes in one direction • Dsh moves opposite of s.e.p; wherever spot of dsh is is where dorsal is • Dsh inhibits gsk3. gsk3 is everywhere in embryo. If dsh is on one side of emrbyo, inhibits gsk3, allowing b catenin to survive. End up with gradient of b cattenin on one side of the egg, and none on the ther • This is all happening before the first cleaveage • By the time gastrulation starts, cell on one side will have beta cetenin, other side wont. Area with high levels of beta catenin forms dorsal • LiCl inhibits gsk3. beta catenin then is allowed everywhere. Embyo becomes dorsalzed, and tries to form dorsal everywhere • UVà put bottom of egg under uv light. Basically depolymerizes and intferes
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• Has not been any transcription yet. • Tcf 3 is found everywhere in the cell. In absence of b-cat, tcf 3, it is a repressor. In presence of b-cat tcf3 causes transcitpion of siamois and twin (found in dorsal) • Don’t necessarily have dorsal organizer yet (need more specificity) siamois and twin turn on transciption of dorsal organizer gene goosecoid.
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• B cat turns on genese with help of tcf 3. turns on siamois and twin, needs smad input. • Smad comes in where only tgfb sugnalling • Neiwkoop center signals to form dorsal organizer • Twin siamosi trun on gooscoid, get dorsal organier • Siamois and twin are really potent. Can cause twinning and ish
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• If have gooseocidà dorsal mesoderm • All properties of dorsal organizer are because of goosecoid • Goosecoid is found in dorsal organizer • If inject goosecoid in cell, will get another organizer • Goosecoid turned on because of niwekwoop signalling undertneath it • Goosecoid is not responsible for signalling, it’s a tf
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• Noggin is found in dorsal roganizer • Makes dorsal emsoderms • Can resuce dorsal axisà if embryo is getting fucked by uv light. If inject noggin, will rescue it. Can make new dorsla organizer. Can inject too much will get a nig head lol (NOGGIN) • If dorsalize embryo will get neural and heaf (no ventr
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• All kind of very similar • Make dorsal things, inhibit ventral things • Chordin found in noochord (so is them all) • . • If take ac cells. Leava lone, get epidermsis. If add tgfb (can induce things like activin) get mesoderm. If dd tdf and follistatin get neural. • All the same, but nodal related is a little different • Rdundnddnacy because making dorsal is important-à dorsal makes neural structures (very necessary)
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• Ecotderm must be induced ot form epideris • If don’t induce ectoderm to form epidermsi, it will form neurl. Forms epidermsis bcause there is bmp. Default of ectoderm is neural, but bmp induces
• Sumamry slide on phone
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• Tgf beta family member, smad signalling • Not all bmp play role in bone • Bmp found in ventral side, gsc in dorsal • If inject bmp everyweherà inhibit dorsal • If inhibit bmp (inject a dm neg receptor for bmp), domain gsc will expand. Basically dorsalizing embryo • Follistatin and chrodin inhbit bmp (also prmote dorsal)
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• Bmp turns on genesà one being msx • Msx1 turns on epidermal genes • Ac cells have bmp which turns on msx • Bmp is maing epidermis and also ventral sturctures (turns on xvent1) • To fomr neural must inhibit bmp turning on msx; noggin, chrodin will take care of this
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• Balance between bmp and dorsal organizer • Two gradients in embryo • High bmp from anterior going to posterior; high noggin going from posterior to anterior • No bmp in posteriorà can form dorsal mesoderm • On other side, cells have bmp, not nogginà leads to ventral structures • Influenced by both form dorsolateral muscle
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• Depending on what you see (bmp, gsc etc), depends on what u become • Xwnt inhibit neural porduction • Bmp makes epidermis, xwnt8 inhibtis neural • Starting gastualtion; give rise t head structures (area is near high bmp and xwnt) • Problem is xwnt inhibts neural, but is found near head
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• Have embryo, dorsla lip • Involuting marginal zone will form dorsal mesoderm • Pharygenal endoderm leads gastulation. Has important molecules tha inhbit xnt. • Ceerbreus promotes extremem anterior strucutres (eyes etc) • Found in leading end of gastrulaion. Don’t signal until reach anterior part • Form neural struactures and inhibts xwnt • Also inhibits bmp • Over express get 2 heads
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• Dickkopfà over express get a bigger head (means stubborn in german) • Ingibits xwnt • Frzbà soluble frizled receptor • Frizzled is receptor fro wnts • On anterior side have wnt; inhibit by binding to wnt receptor • Frzb binds to wnts and remove them from the signal pathway • To form epidermisà need bmp and wnt wprking • To form spinal chordà need to inhibit bmp (nogging chrodin and follastatin) • To form head and neauralà need to inhibit bmp and wnt
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1
• When gastrulation starts, has tissue above dorsal lip • Organizer is comitted, gives rise to dorsal mesoderm • If you put the organizer elsewhere get 2 dorsals • Dorsal organizer is set up opposite sperm entry point • First cleavage plane defines left and right, goes through the organizer • Grey crescent develops; tells where dorsal is because of crodical rotation • If divide embryo and don’t give it grey crescent, develops abnorally • If ectoderm is transplanted before it is indeced, it will remian ectoderm • If wait unitl gastrulation prgresses; ectoderme is being induced and changed(knows position is neural). If transplant to another embryo will form neural tissues
2
• Red band forms mesoderm • If take ac and vp, and put them together, will stell get signalling and form mesoderm (wont get any patterning) • PCR for mesodermal genes. If take late state change embryo that has a heart, will have the gene (cardiac actin). Wont have any in ac, wont have them in vp. However if u put ac and vp together, may get cadiac actin (induced the right type of actin)
3
• Organizer is responsible for 1o embryonic inductuion • 4 function: 1. Make something called the prechordal plateà self determing, forms this all by itself 2. Starts off as ring of mesoderm. Notochord (dorsal mesoderm). The closer u are to them, the more dorsal character you are. Its whats maing somites dorsal. 3. Makes ectoderm near it neural, because of signalling 4. Intiates gastrulation
4
• Cortucal rotationà rotates respect to inside of egg • Does so because of mt lining cortex, outside of egg • Before fert, mt will not be organized, after fert mt much better organized, soerm bring centriole • Centriole of mt organized centrer; beofre centrioles play role in mt apparatus. Centriole is mtoc allows mt to polmerate away from sperm that just came in • Provides track • When it rotates, cytoplasm floats around. When it swirls, able to see the grey crescent (dorsal side). First cleavage goes theough the grey crescent
5
• Dsh is on vegetal side in cortex • Sperm ferts in animal half. When fert brings in centriole. When centriole comes in; polymerizes green tract. Causes rotation of the cortex (causes things insde to swil around), also gives dsh a tract to move. Kinesin moves dsh to move. • Polymerization downwards in respect to gravity. If gravity isn available polymerizes in one direction • Dsh moves opposite of s.e.p; wherever spot of dsh is is where dorsal is • Dsh inhibits gsk3. gsk3 is everywhere in embryo. If dsh is on one side of emrbyo, inhibits gsk3, allowing b catenin to survive. End up with gradient of b cattenin on one side of the egg, and none on the ther • This is all happening before the first cleaveage • By the time gastrulation starts, cell on one side will have beta cetenin, other side wont. Area with high levels of beta catenin forms dorsal • LiCl inhibits gsk3. beta catenin then is allowed everywhere. Embyo becomes dorsalzed, and tries to form dorsal everywhere • UVà put bottom of egg under uv light. Basically depolymerizes and intferes
6
• Has not been any transcription yet. • Tcf 3 is found everywhere in the cell. In absence of b-cat, tcf 3, it is a repressor. In presence of b-cat tcf3 causes transcitpion of siamois and twin (found in dorsal) • Don’t necessarily have dorsal organizer yet (need more specificity) siamois and twin turn on transciption of dorsal organizer gene goosecoid.
7
• B cat turns on genese with help of tcf 3. turns on siamois and twin, needs smad input. • Smad comes in where only tgfb sugnalling • Neiwkoop center signals to form dorsal organizer • Twin siamosi trun on gooscoid, get dorsal organier • Siamois and twin are really potent. Can cause twinning and ish
8
• If have gooseocidà dorsal mesoderm • All properties of dorsal organizer are because of goosecoid • Goosecoid is found in dorsal organizer • If inject goosecoid in cell, will get another organizer • Goosecoid turned on because of niwekwoop signalling undertneath it • Goosecoid is not responsible for signalling, it’s a tf
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• Noggin is found in dorsal roganizer • Makes dorsal emsoderms • Can resuce dorsal axisà if embryo is getting fucked by uv light. If inject noggin, will rescue it. Can make new dorsla organizer. Can inject too much will get a nig head lol (NOGGIN) • If dorsalize embryo will get neural and heaf (no ventr
10
• All kind of very similar • Make dorsal things, inhibit ventral things • Chordin found in noochord (so is them all) • . • If take ac cells. Leava lone, get epidermsis. If add tgfb (can induce things like activin) get mesoderm. If dd tdf and follistatin get neural. • All the same, but nodal related is a little different • Rdundnddnacy because making dorsal is important-à dorsal makes neural structures (very necessary)
11
• Ecotderm must be induced ot form epideris • If don’t induce ectoderm to form epidermsi, it will form neurl. Forms epidermsis bcause there is bmp. Default of ectoderm is neural, but bmp induces
• Sumamry slide on phone
12
• Tgf beta family member, smad signalling • Not all bmp play role in bone • Bmp found in ventral side, gsc in dorsal • If inject bmp everyweherà inhibit dorsal • If inhibit bmp (inject a dm neg receptor for bmp), domain gsc will expand. Basically dorsalizing embryo • Follistatin and chrodin inhbit bmp (also prmote dorsal)
13
• Bmp turns on genesà one being msx • Msx1 turns on epidermal genes • Ac cells have bmp which turns on msx • Bmp is maing epidermis and also ventral sturctures (turns on xvent1) • To fomr neural must inhibit bmp turning on msx; noggin, chrodin will take care of this
14
• Balance between bmp and dorsal organizer • Two gradients in embryo • High bmp from anterior going to posterior; high noggin going from posterior to anterior • No bmp in posteriorà can form dorsal mesoderm • On other side, cells have bmp, not nogginà leads to ventral structures • Influenced by both form dorsolateral muscle
15
• Depending on what you see (bmp, gsc etc), depends on what u become • Xwnt inhibit neural porduction • Bmp makes epidermis, xwnt8 inhibtis neural • Starting gastualtion; give rise t head structures (area is near high bmp and xwnt) • Problem is xwnt inhibts neural, but is found near head
16
• Have embryo, dorsla lip • Involuting marginal zone will form dorsal mesoderm • Pharygenal endoderm leads gastulation. Has important molecules tha inhbit xnt. • Ceerbreus promotes extremem anterior strucutres (eyes etc) • Found in leading end of gastrulaion. Don’t signal until reach anterior part • Form neural struactures and inhibts xwnt • Also inhibits bmp • Over express get 2 heads
17
• Dickkopfà over express get a bigger head (means stubborn in german) • Ingibits xwnt • Frzbà soluble frizled receptor • Frizzled is receptor fro wnts • On anterior side have wnt; inhibit by binding to wnt receptor • Frzb binds to wnts and remove them from the signal pathway • To form epidermisà need bmp and wnt wprking • To form spinal chordà need to inhibit bmp (nogging chrodin and follastatin) • To form head and neauralà need to inhibit bmp and wnt
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