Etchevers:Notebook/STRA6 in eye development/2008/09/05

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Stem cell marker expression in the cornea

Just for future reference, downloaded an article showing Abcg2 expression in the mouse brain during E13 (Theiler 21), E18ish (Theiler 25) and postnatal day 3. The reference came from Visigene.

So can perhaps use a P1 telencephalon, in particular a transverse section through the olfactory bulbs underlying it, for a + control for Abcg2 and Pax6 expression. Jackie has successfully synthesized probes for these, from the mouse Pax6 probe provided us and validated by Fabienne Pituello from Peter Gruss before that, I think, in pBS. We designed primers to amplify just the insert from the plasmid and add on T7 sequences for RNA synthesis.

The Abcg2 yield was weak for both S and AS in dot blot, and for all in spectrophotometer dose - 150 to 200 ng/ul. Sufficient for hybridization though. Stephane is trepanating mouse corneas today, will fix both unoperated and scarring corneas to confirm the published results about stem cells among the keratocytes and to see if they are upregulated in the damaged corneas. Will fix some in Formoy (modified Carnoy's) solution for paraffin embedding and in situ hybridization, and some fresh-frozen for the cryostat for immunohistochemistry.

  • Alethea 07:49, 5 September 2008 (EDT):