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lec09_morg_bell

lec09_morg_bell - Extrinsic and intrinsic factors control...

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Extrinsic and intrinsic factors control the genesis of amacrine and cone cells in the rat retina Michael J. Belliveau and Constance L. Cepko (1999) Presented by Bo Morgan on March 7, 2004.
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Introduction Progenitor Cells Postmitotic Cells Do postmitotic cells influence progenitor cell fate deci- sions? Secreted factors shown to stimulate or inhibit rod cell development. Non-neuronal cells show feedback mechanisms from fated cells to the stem cell population.
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Why focus on the retina? Evidence for particular order of production of cell types Evidence for particular portions of cell types Diagram removed for copyright reasons.
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How is prenatal retina progenitor cell fate affected by different surrounding cell populations? Progenitor cells Explant with extract culture Dissociate and reaggregate culture Low density culture E16 explant Assay for cell type Which retinal cell types do the progenitor cells ultimately become? Three types of culture preparation methods: 1. Dissociate and reaggregate with retinal postnatal cells 2. Explant to culture with postnatal retinal extract 3. Put progenitor cells into low density culture before mixing with postnatal cells.
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Methods Overview P0 retinal extract applied P0 and E16 Dissociation of cultures Explant cultures E16 tissue Pulsed 1 hour for timing E16 cells H thymidine labeling 3 P0, E16 and P0:E16 Reaggregation of cultures Establish t 0 Create 2 controls and a 20:1 mixture Closer to in vivo situation [ 3 H ] thymidine labeling of E16 cells to select progenitor population Dissociation and Reaggregation of cultures (E16, P0 and E16:P0) 2 control solutions (E16 and P0) and 1 mixture solution (E16:P0) are prepared.
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[ 3 H ] thymidine labeling of prenatal cells B. [ 3 H ] thymidine-positive: cells dividing during [ 3 H ] thymidine pulse C. DAPI-positive: cell nuclei D. VC1.1-positive: amacrine cells Source: Belliveau, M. J., and C. L. Cepko. "Extrinsic and Intrinsic Factors Control the Genesis of Amacrine and
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