Test 3 Flashcards
As a zygote, the organism:
- Consists of one cell of only one type
- Consists of unrealized genomic potential
- Operates using stored materials
- Has “relationships” with no other cells
- Is relatively homogeneus
True or false, the zygote is polarized to some degree
True
What does early development involve
- Development of new “raw materials” for building embryo
- Establishing new relationships between numerous cells
- Further polarizing the organism
Steps of early development
- Cleavage
- Establishment of cell fate
- Establishment of body axes
- Gastrulation
Cleavage
- Cell cycle control
- Rapid cell divisions, division of fertilized egg into many cells
Establishment of cell fate
Specification
Establishment of body axes
(anterior posterior, dorsal ventral, left right)
Gastrulation
Cells undergo displacement, cells move to different parts of the embryo, cells aquire new neighbors
How are cell fates specified during cleavage
Cell to cell interactions and asymmetric distribution of morphogenic determinants
What initially controls cleavage
Factors stored in the egg
Factors stored in the egg
Stored proteins, mRNAs
Mitosis promoting factor
Stimulates the cell cycle (initiates cell division during mitosis and meiosis)
Where does MPF come from
Mother
What stops division
Absence of MPF
Cyclin B
Controls cdc2 activity
cdc2
Cyclin-dependent kinase
CDK
Phosphorylates histones
What happens when cyclin B degrades
Cell division stops
What controls the cyclin B presence and degradation
Egg cytoplasmic proteins
What happens when the maternal stores are used up
Embryonic control of the cell cycle
Mid Blastula Transition
Must produce its own proteins
Post MBT
- Cell cycle adds two new G phases
- New mRNA transcription
- Cell division becomes asynchronous
How does the cell cycle of blastomeres compare with that of somatic cells
Cycle is like somatic cells after MBT
Karyokinesis
Mitotic division of cells nucleus
Cytokinesis
Division of cytoplasm plus organelles
Mechanical agent of karyokinesis
Mitotic spindle
Mechanical agent of cytokinesis
Contractile ring
Major protein composition of karyokinesis
Tubulin microtubules
Major protein composition of cytokinesis
Actin microfilaments
Location of karyokinesis
Central cytoplasm
Location of cytokinesis
Cortical cytoplasm
Positioning of mitotic spindle and contractile ring relative to eachother
Perpendicular
What affects how cleavage occurs
Position of centrioles and yolk
Types of cleavage
- Isolecithal
- Mesolecithal
- Telolecitithal
- Centrolethical
Isolecithal
Little yolk, division throughout
Mesolecithal
Medium yolk, more division on top
Telolecithal
Large yolk, almost all division on top
Centrolethical
Division in random spots
First cleavage
Meridonial
Second cleavage
Meridonial
Third cleavage
Equitorial
Fourth cleavage
Unequal cleavage between animal and vegital hemispheres
Micromeres
Smallest cells
Macromeres
Biggest cells
Mesomeres
Medium cells
How do the animal and vegetal halves differ
Cells on the bottom have different morphogens than on top
7th plus division
At 128 cells, divisions become less regular. Forms blastula
How many cells thick is the blastula
One (hollow inside)
What do the cells of the blastula adhere to
- Hyaline layer (outside)
- Basal membrane (inside)
- Neighboring cells
What develops on the blastula
Cilia
What is initiated at the animal pole of the blastula
Hatching (hatches out of fertilization envelope)
What changes the osmotic pressure of the blastula
Sodium influx
What fills the cavity of the blastula
Proteinaceous fluid fills cavity
What expands the blastocoel
Osmotic pressure
Mechanisms for establishing asymmetry (in regards to splitting cytoplasmic determinants)
- Patterning molecules bound to egg cytoplasm
- Molecules actively transported along the cytoskeleton (made of microtubules in secretory vesciles)
- Molecules become associated with one centrosome, and then follow that centrosome into one of the two mitotic sister cells
What happens when asymmetry is established
One cell can specify another and participate in reciprocal inductions
Notochord
Induces spinal cord formation
Where is the animal pole located
Top
Mesenchyme
Loosely packed, unconnected, mesoderm, neural crest origin
Coelom
Internal body wall, mesoderm origin
What type of specification do micromeres undergo
Autonomous
60 cell stage
Specified but not committed. Blastomeres mostly undergo conditional specification
What cells trigger early induction events
Micromeres
What do micromeres have enough information to do
Induce recognizable larvae and secondary axis
____ plays a role in micromere specification
B-catenin
What activates B-catenin
Wnt signal transduction pathway
What cell fates does B-cateni accumulate in
Endoderm and mesoderm
What do veg2 cells with B-catenin become
Endomesoderm
______ causes accumulation of B-catenin in every cell
LiCl, transforms presumative ectoderm into endoderm
What will animal cells become if they have B-catenin
Endoderm
What micromere protein activates the notch pathway in adjacent veg2 cells
Delta
What does notch pathway activation result in
Secondary mesanchyme
What is Wnt8 made of
Micromeres and endoderm cells
What signal are endomesoderm cells not recieving
Delta
What type of signal does Wnt8 act as
Autocrine signal
What does Wnt8 boost specification in
Veg2 endoderm and micromeres
Invagination
Infolding region of the cells
Ingression
Migration of individual cells from the interior surface layer into the interior of the embryo (cells become mesencymal and migrate independently)
Involution
In turning or inward movement of an expanding outer layer spreads over the internal surface of the external cell layer
Delamination
Splitting one cellular sheet into two parallel sheets
Epiboly
Epithelial sheets spread and enclose deeper layers of the embryo
Intercalation
Two rows become one
How does ingression work
Cells lose cadherin and migrate up the blastocoel to a spot
What gradients move cells in ingression
VEGF and FGF (growth factors)
What does invagination of the vegetal plate form
Archenteron (primative digestive tract)
How does the forgut move to the top
Invagination, involutes inwards
What appears to be responsible for the initial invagination that occurs during gastrulation
Osmotic gradient
How are cells freed from the hyalin layer to be able to move upward
Changing osmotic gradient
What do cells do during formation of the archenteron
Cells intercalate and move forward at the same time (results: thin tube)
When is the anterior posterior axis formed
Very early on with the animal vegetal axis
What does the vegetal region have that is necessary for posterior development
Maternal components
When is the dorsal ventral axis established
After fertilizatio
Another name for the dorsal ventral axis
Oral-aboral
Where is the dorsal ventral axis
Approx. 45 degrees clockwise from the first cleavage plane
What promotes oral fates
Nodal
What promotes aboral fates
BMP2/4
When is the left right axis established
After oral-aboral
The left right axis has ____ expression of the nodal gene
Asymmetric
What sets up the axis in amphibians
The organizer
What is the organizer analogous to in sea urchins
Mircomeres
Inductions of the organizer
- Dorsal ventral axis
- Mesoderm
_Ectoderm
Where does fertilization occur
Anywhere on the animal pole
Cortical rotation
Contents in egg shift
Where are cytoplasmic determinants shifted to
Gray crescent
Where is the gray crescent located
Directly diagonal to the site of sperm entry
How many degrees does the egg rotate
30
What does the first cleavage plane cut in half
Gray crescent
Where is the dorsal located
Opposite sperm entry
Where does gastrulation begin
Gray crescent
What does cortical rotation relocate
Maternal factors
What is the dorsal most and vegetal most region
Nieukoop Center `
Yolk classification of amphibians
Mesolecithal
What pole is the yolk in for amphibians
Vegetal pole
Where do more cells develop in the amphibian embryo
Animal pole
What type of cleavage occurs in amphibians
Displaced radial
Holoblastic
Complete cleavage
Meroblastic
Incomplete cleavage
Is amphibian cleavage holoblastic or meroblastic
Holoblastic
Major functions of blastocoel
- Permits cell migration during gastrulation
- Prevents cells beneath it from interacting prematurely with cells above it (cells on top are different from bottom cells)
What type of movement forms the archenteron
Invagination
Beginning of frog gastrulation
Formation of dorsal lip
Vegetal rotation
Invagination of bottle cells
Involution of marginal zone cells
Dorsal blastophere lip
Site where gastrulation begins (cells stream through the opening)
What happens to the blastocoel during gastrulation
Will eventually go away and be replaced
What does gastrulation form
3 germ layers
Where is archenteron located
Blastopore
What is the yolk plug
Spot not yet covered
What are axes specified by
Events triggered at fertilization and realized during gastrulation
What is the mesoderm determined by
Transcription factors and paracrine factors from the vegetal region
What is the only tissue in the xenopus pre-gastrula that has its fate determined
Dorsal lip of the blastopore
What will the dorsal lip determine the fates of
Notochord and head endomesoderm
What decides if it will be ventral or dorsal mesoderm
Nodal
Both cells have equal amount of gray crescent
Normal development
One cell has entire gray crescent
Normal development of one, only belly piece in other
Gray crescent
Future dorsal lip of the blastopore (future site of gastrulation)
Transplantation in early gastrula
Conditional specification
Transplantation in late gastrula
Autonomous specification
Spemann and Mangolds experiment
Dorsal blastopore transplanted to another embryo, secondary axis forms
What is the conclusion of the spemann and mangold experiment
The dorsal lip is the organizer
What does the organizer do in spemann mangold experiment
- induces the host ventral tube and dorsal mesodermal tissue such as somites
- organized host and donor tissue into secondary embryo with a clear a-p and d-v axis
- organize dorsal ectoderm into neural tube and transform flaking mesoderm into anterior posterior body axis
Induction
Signal from one group of cells
The mesoderm is induced to be dorsal, intermediate or ventral based on _______
Location
Dorsal signal
B-catenin
B-Catenin
- anchor for cadherins
- nuclear transcription factor in wnt
B catenin in sea urchins
specifies micromeres and endomesoderm
B catenin in xenopus
specifies dorsal structures
Where is B catenin initially
Spread throughout the embryo
Where does B catenin become accumulated in
Dorsal cells
Where is B catenin concentarted
Nieukoop center and organizer
Dorsalization of B catenin accomplished by
- Protecting B catenin in the dorsal area
- Degrading B catenin anywhere else
What causes accumulation of B catenin only on dorsal side of the embryo
Cortical rotation
What happens to B catenin without Wnt
It is broken down
GSK3 distrubution
Distributed throughout the embryo
What marks B catenin for degradation
GSK3
Dishevled (Dsh)
Blocks GSK3 activity
Where is Dsh localized
Cytoplasmic cortex at the vegetal pole
GSK3 present
No B catenin, ventral
No Gsk3
B catenin present (Dsh blocked GSK3), dorsal
Dsh present
B catenin present, dorsal
GBP
GSK3 binding protein
Kinesin
Motor protein
How is Dsh moved
Moved on microtubules. Hitches a ride on GBP. Kinesin helps it move
Where is beta catenin present after GSK3 acts
Marginal area opposite the point of sperm entry (ie. future dorsal lip)
What transcirption factor does B catenin act with
Tcf3
What do B catenin and Tcf3 do
Stimulate expression of dorsalizing genes
Dorsalizing genes
Siamois and Twin proteins, Goosecoid and Xlim1
Siamois and Twin
Tracription factors to turn on more dorsalizing genes
Gooseoid and Xlim1
Specify dorsal mesoderm
B catenin pathway (organizer induction)
B catenin –> nodal related high –> organizer (dorsal)
VegT, Vg1 pathway (mesoderm or organizer induction)
VegT, Vg1 –> nodal related high –> organizer (dorsal)
OR
VegT, Vg1 –> nodal related low –> ventral mesoderm
Summary of mesoderm induction
- Maternal RNAs tethered to the vegetal cortex
2. Gardient of nodal (TGFB family protein) signal
Maternal RNAs thethered to vegetal cortex
- Vg (TGFB family protein)
- VgT (transcription factor)
Role of VgT
Instruct endoderm to express another TGFB family activin, derriere and nodal
- Induce to express Xbra or goosecoid
- induce mesodermal development
Gradient of nodal signal
Nodal gradient is formed by expression of VgT and B-catenin in endoderm
What does nodal gradient induce
Mesoderm formation
Less nodal
Ventral mesoderm
Intermediate nodal
Lateral mesoderm
Highest nodal
The organizer (dorsal)
Functions of the organizer
- Can self differentiate dorsal mesoderm
- Can dorsalize the surrounding mesoderm into paraxial mesoderm
- Can initiate the movement of gastrulation
- Can dorsalize the ectoderm to induce neural tube
What does the organizer develop into
- Pharangyeal ectoderm (frontal ectodermal cells)
- Head mesoderm (frontal mesodermal cells)
- Dorsal mesoderm (notochord)
- Dorsal blstopore lip
What does anterior posterior axis development come after
Dorsal ventral axis development
What do the first migarting endomesodermal cells become
Anterior part
What do later migrating mesoderm cells become
Ventral lips (form posterior structures)
Organizer inductions
- Mesoderm
- Dorsal/ventral axis
- Ectoderm and neural tissue
What does ectoderm usually want to become
Neural tissue
How does ectoderm become epidermis
Needs signa (BMP)
BMP inhibitors
Noggin, Chordin, Follistatin
What activates BMP inhibitors
Smad 2/4 and Siamois/Twin
What do BMP inhibitors do
Stop BMP and make neural tissue
Where are BMP inhibitors located
Dorsal side
Where is more BMP located
Ventral side
What happens as levels of noggin increase
More neural structures develop
Nuclear proteins
Remain within the cell
Where are noggin, chordin and follistati found
Organizer
Epidermal inducers
BMP4, BMP2, BMP7
Job of BMPs
Repress genes involved in forming neural tissue while activating other genes involved in epidermal specifiaction
Add BMP to embryp
No dorsal structures, no neural tube
Blocked BMP in embryo
Spread to ventral side and form more neural tissue than they were supposed to
Proneural gene
Neurogenin
Making sure parts of the brain are made in the right spot
Regional specification of the neural structures that are produced
FrzB
Binds to Wnt and prevents it from binding to receptor on cell and activating it (acts as receptor)
What is FrzB secreted from
Organizer (dorsal)
Where is Wnt secreted from
Ventral
FrzB blocks _____ from dorsal region
Xwnt8
When is d-v axis set up
At ferilization
When is a-p axis set up
Established by gatsrulation movements across the dorsal lip of the blastopore
L-R axis establishment
Nodal expression on left, not righrt
Gene that expressed eft right axis
Xnr1
How is xnr1 expression limited to the left side
Process involving cortical rotation and Vg1
What happens if Xnr1 is blocked
Random gut coiling and heart looping
What direction does the heart loop (normal)
Left
What direction is gut coiling (normal)
Counter clockwise
What happens if nodules are mutated
No left or right axis
Yolk amount in mammals
Isolethical
How do mammals gastrulate
Gastrulate as if there is a lot of yolk `]
What is the blastula called in mammals
Blastocyst
When is the first mammalian cleavage
1 day after fertilization
How far apart are cleavages in mammals
12-24 hrs
Cleavage type mammals
Rotational
Rotational cleavage
Not happening with all cells at the same time, no MBT
Compaction
8 cell stage, tight junctions between outside cells (e-cadherin), seal off inside of sphere
What will ICM form
Embryo proper
What will trophoblast form
Extraembryonic structures
What secretes hormones to cause the uterus to retain the fetus
Trophoectoderm
Cavitation
Trophoblast secretes fluid into the morula (with Na+ pumps, `creates blastocoel and pushes ICM to one side)
Zona pellucida
Prevents adhesion to uterine wall
Why does adhesion to the uterine wall need to be prevented
Must hatch out for adhesion, premature adhesion = eptopic pregnancy
What is formed when trophoblast attaches to the uterine wall
Chorion
Chorion
Embryonic portion of the placenta
What is secreted by the trophoectoderm and why
Proteases
- Digests uterine ECM
- Blastocyst implants
What does ICM form besides embryo proper
Yolk sac, allantois, amnion
What does the allantois turn into
Part of the bladder
Which is a stem cell, ICM or trophoectoderm
ICM
Cells of the blastocyst
ICM and trophoectoderm
What does ICM become
Epiblast and Hypoblast
Epiblast
Forms embryo proper
Hypoblast
forms extraembryonic membrane - yolk sac
What are the epiblast and hypoblast together callled
Bilaminar germ disc
Cytotrophoblast
- adheres to endometrium
- proteolyze uterine wall
- secretes paracine factors to attract maternal blood vessels
- displaces vascular tissue
Syncytiotrophoblast
Digests uterine tissue (move further into uterine wall)
Extraembryonic endoderm
Gives rise to the yolk sac
Extraembryonic mesoderm
Gives rise to blood vessels and umbilical cord
Job of paracrine factors
Recruit maternal blood vessels towards embryo
Decidua
Maternal portion of the placenta
What does the epiblast become
Amniotic ectoderm or embryonic epiblast
Embryonic epiblast division
Embryonic ectoderm of primative streak (which will become embryonic endoderm or embryonic mesoderm, sometimes extraembryonnic mesoderm)
What does the hypoblast become
Hypoblast –> extraembryonic endoderm –> yolk sac –> (extraembryonic mesoderm sometimes)
What does the trophoblast become (pathway)
Trophoblast –> cytotrophoblast –> syncytiotrophoblast
What does the chorion surround
Embryonic membrane
What is the cloud around the fetus
Blood vessels, part of the chorion
Cells that form yolk sac
hypoblast, blastodisc cells
Function of yolk sac
No nutritional function (vestigial), important for blood cell formation
What is the amnion formed by
Epiblast… ectodermal and mesodermal cells complete amnion
Amniotic fluid purpose
Cushions embryo or fetus
Allantois formation
Begins as outpocket of endoderm near yolk sac base, endodermal and mesodermal cells form stalk attaching to blastocyst wall
Purpose of allantois before it becomes the bladder
Vestigial respiratory organ
What is the chorion formed from
Mesoderm and trophoblast near allantois
How does the chorion get nutrients
Chorion vili invade endometrium
Placenta purpose
Primary embryo support, oxygen and nutrients exchanged for carbon dioxide and wastes
What do umbilical arteries carry
Deoxygenated blood from fetus
What do umbilical veins carry
Oxygenated blood to fetus
Early twin separation
2 chorions, 2 amnion
Middle twin separation
1 chorion, 2 amnion
Late twin separation
1 chorion, one amnion
Evidence that ICM generate any cell type
Twin formation (ICM split and both halves can become a whole embryo)
Where does human gastrulation begin
Primative streak
Movement of primative streak
Posterior to anerior, back posterior and disappears
What is the primative streak analogous to in amphibians
Blastopore (where movement first starts)
Ectoderm
Nonmigrating superficial blastodisc cells
Endoderm
Cells facing yolk sac
Mesoderm
Poorly organized cells between two other germ layers
What day does gastrulation occur in humans
12-15
What germ layer is created first in humans
Endoderm
Where will the brain form
Neural plate
How does the neural tube form
Primative streak loops
Where does the neural plate form
In the ectoderm along embryo posterior
What forms CNS cavities
Neural folds
Week 3
Head fold
Chorionic vili
Heart starts beating
Allantois forms
Week 4
Body stalk
Yolk stalk
Other organs form
Body stalk
Between the embryo and chorion. Forms carrying blood to and from the placenta
Yolk stalk
Narrow connection between endoderm and yolk sac
When does embryonic folding occur
4 weeks
Week 5
Umbilical stalk
What forms the umbilical stalk
Body and yolk stalks
Week 6
Brain waves, mouth and lips, fingernails, limbs lengthen, skull bones form
When does it become a fetus
9 weeks
Week 10
Bones replace cartilage. Fetus is connected to placenta by umbilical cord
Amniotic cavity fills uterine cavity
What is the umbilical cord made of
Allantois, blood vessels, remnants of yolk stalk
Carnegie stages
Stage based on level of development, regardless of time passed
Anterior posterior patterning mammals
BMP, Wnt, RA, Hox genes
Dorsal ventral patterning mammals
Mystery
Left right patterning mammals
Cilia, Hedgehog, RA, FGF
Mammalian organizer
AVE and Node
AVE
Anterior visceral endoderm, secretes anterior markers
Node
(same as spemann organizer in amphibians), creates all body and neural features
Anterior patterning
BMP and Wnt antagonists (some RA)
Posterior patterning
Wnt, BMP, FGF, RA
Retinoic acid gradient
Posterior –> high, anterior–> low
How is the anterior determined
Blocking posterior signals
What determines A-P axis patterning
Hox genes
What activates hox genes
Cdx1,2,4
What activates cdx family
RA, Wnt3a, fgf8
Homeotic selector genes
Determine segment identity
Where are hox genes expressed
Along the dorsal axis
How is the level of body along the A/P axis determined
Most posterior hox gene expressed
What does hox knockout do
Shifts vertebrae
What are hox genes sensitive to
RA
Direction of RA gradient
P-A
Where does hypoblast form
Side of ICM exposed to blastocyst fluid
Where does dorsal axis form
From ICM cells in contact with trophoblast and amniotic cavity
Levels of regulation for left right
Organ specific
Global
Organ specific
IV gene
Mutations cause randomized LR asymmetry for each organ, can be fatal
Global
Inversion of Inv gene
Mutations cause asymmetrical organs to be reversed, usually not a large problem
Active genes on left side
Nodal and Pitx2
LR mechanism frog
Vg1 placement
LR mechanism chick
Suppression of sonic hedgehog
LR mechanism mouse
Asymmetirc distribution of Shh
Dynein
Motor protein
What cells are ciliated for LR
Node
NVP
Nodal vesicular parcels
Contain Shh and RA
What happens if parcels are not secreted
LR asymmetry fails to establish
What are cilia powered by
Dynein and ATPase
What do iv genes code for
Dynein protein