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TGF-beta pathway Roles
important for embryonic patterning, key roles in embryonic stem cell self-renewal, gastrulation, differentiation, organ morphogenesis.
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What is BMP
- subfamily of TGF-beta pathway.
- Required for NEURAL CREST signaling
- Regulated by SNW1: this act to allow for NEURAL crest formation
- knockdown of SNW1 inhibits neural crest formation
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Role of FGF signaling
- Works via RTK pathway
- Responsible for limb bud initiation and outgrowth
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Shh
- Responsible for limb bud FORMATION
- Aids in patterning at AER by production of FGFs
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Glomus cell markers
- BrdU
- Th-T is also found in glomus cells
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What is responsible for neural pruning
CASPASE activity: aids in dendrite removal (DRONC is the initiator caspase)
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Epigenetics
- Changes in genetic REGULATION without a change in the DNA sequence
- Changes due to METHYLATION of promoter region, and HISTONE MODIFICATION
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Role of REST
- Marker for embryonic stem cells (ES) REGULATES transition from pluripotent to NEURAL stem/progenitor cells and from progenitor cells to MATURE Neurons
- REPRESSES NEURAL gene activity, until needed: then Dissociates from RE1 site in order to ACTIVATE neural genes
- *REST is Gone during RA treatment and differntiation into neurons
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Neuronal Marker and RA
- Neural marker TuJ1: expressed in NEURONs
- Retinoic acid: Aids in transition to promote differentiation into NEURONS
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Pluripotency
- the ability of stem cells to differentiate into any of the three germ layers:
- - Endoderm (stomach lining, lungs)
- - ectoderm (epidermis and nervous system)
- - mesoderm (muscle, bone, blood, etc)
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How would you determine if cells are pluripotent?
- Use RT-PCR for specific genes involved in induction of pluripotentcy: Sox2, Oct3/4 (TFs)
- If when injected (retrovirally) and looked at a microarray..and look for similarities to embryonic stem cell (ES cells), which they are PLURIPOTENT
- Also use immunohistochemistry to look at specific markers for cell lineages
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Comparing pluripotent cells to stem cells using what experimental approaches
- RT-PCR to determine presence (or absense) of specific genes,
- Similarly with DNA microarrays
- CHIP assays ??????Explain
- Global gene expression analysis: visualized using scatter plot, with regions closely resembling each other on or within the two linear slope lines
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