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What is the ECM made of (3 things)?
Collagen, Fibronectin, laminin
What are the three major effects of mitogens/growth factors?
Induce transcription of cyclins and cdks, block expression of cdk inhibitors, induce rapid phosphorylation of Rb
What is Rb?
Retinoblastoma gene, the phosphorylated Rb allows cycle cycle progression past the G1 restriction point
fibroblast growth factor: growth factor(cytokine) that interacts with tyrosine phosphate receptor
transforming growth factor beta. major stimulus for fibrosis
epidermal growth factor. interacts with tyrosine kinase receptor.
vascular endothelial growth factor. required for vasculogenesis
platelet-derived growth factor. mitogen for smooth muscle, derives macs and neutrophils towards it
Is cell growth and cell proliferation the same?
How do you measure DNA synthesis?
3H-thymidine incorporation
How do you measure DNA content?
FACS, fluorimetry
How do you measure cell cycle transit?
proliferation cell nuclear antigen, ki67
How do you measure cell number?
Metabolic dyes, counting cell number (hemocytometer, coulter counter)
How do you measure protein synthesis?
33S-Methionine incorporation, total protein levels
How do you measure cell size?
What is divisional assymetry?
When a stem cell splits into two daughter cells, one inherently contains something different while they other may differentiate or becomes something different
What is terminal differentiation?
When a cell reaches \"maturity\" and can no longer differentiate
What is commitment?
It is the irreversible progression from a stem cell to a particular lineage
What are epigenetic modifications?
Heritable changes in gene expression but not changes in DNA sequence
What is the stem cell niche hypothesis?
Signals from the cell microenvironment directs the cell fate
What are the three components of the stem cell niche?
secreted molecules, cell-cell interactions, integrin-extracellular matrix interactions
What are three ways to perform transcriptional control?
histone modifications, DNA methylation, transcription factors
What are three ways to perform post-transcriptional control?
RNA turnover, alternative splicing, microRNAs
What are two ways to perform post-translational control?
phosphorylation, gylcosylation
Where do post-translational modifications occur?
Histone tails, mostly
What are two types of modifications that can be done to histone tails?
Acetylation and methylation
What do microRNAs do?
inhibits translation of target mRNAs
What are integrins?
Cell adhesion molecules that mediate cell-matrix interactions, signals from inside to out
How is apoptosis different from necrosis
Apoptosis - programmed, cell shrinks, necrosis - trauma, cell explodes
What are four characteristics of apoptosis?
cell shrinkage, chromatin condensation, cytoplasmic blebbing, nuclear fragmentation
What are caspases?
cytoseine proteases that cleave following aspartate. activates apoptosis
What do caspases inactivate?
DNA polymerase, cytoskeletal proteins, nuclear membrane, transcription factors
What do caspases activate?
What is FasL
Fas Ligand, released from inflammatory cells or cells following trauma
What is Bcl2/bclxl?
anti-apoptotic: preseves mitochondrial integrity
What is bax/bad?
prop-apoptotic, disrupts mitochondrial integrity
What is cytochrome c?
Protein that is normally found between the inner and outer mitochondrial membrane, release when undergoing apoptosis
What is Apaf 1?
A protein present in the cytosol that can bind to cytochrome c, this then activates the caspase cascade
what is anoikis?
apoptosis induced by loss of anchorage
What do integrins integrate?
ECM and the intracellular cytoskeleton
Why are integrins important for survival
tells the cell that it is anchored and should survive
What is nuclear morphology for?
measuring cell death, would be fragmented
What is TUNL for?
measuring cell death, present with apoptotic cells
What is Annexin V used for?
measuring cell death, detects protein that is on inner part of membrane and it\'s outer
What is PARP used for?
measuring cell death
What does calcein AM do?
stain live cell cytoplasm green
What does ethidium homodimer-1 do?
stains dead cell nuclei red

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