Both cell proliferation and migration contribute to reepithelization of wounded skin. To observe the effects of any genes or compounds on cell migration, we usually use the scratch assay to assess the keratinocyte motility, and regard it as a measure for the cell migration in vivo. Normally, in the scratch assay we treat the cells with mitomycin c to block the cells' proliferation. Can we treat mice with any small molecular inhibitors to block the proliferation of keratinocytes so that we may assess the cell migration in vivo? If this is not possible, are there any other methods that can be applied for this purpose?
I would think that an inhibition of the proliferation in the mouse might dramatically alter its health and therefore the regeneration.
To assess the participation of cell proliferation in vivo I would rather use EdU/BrdU injections or do some Ki67 immunostainings.
Concerning the migration, I would probably try the reporter mice. If you have a specific cell target, you should use the Cre-lox system with a Cre ensyme under the control of a gene specific o your targeted cells crossed with a reporter strain (beta-galactosidase, tomato, GFP, Confetti, etc). It is a bit a hassle to analyse, but the results are usually beautiful!
Thank you, Thibault! I had just registered in the Researchgate; and this is actually my time to a question here. Your kind reply really encourages me.
Yes, generally speaking, inhibtion of proliferation in the mouse could likely impair its health. I was thinking about blocking the proliferation in the specific context, skin wounding, where some special mechanisms/pathways are involved in regulating cell proliferation. Thus, at least within a limited span of time (for example, 5~7 days), to repress these pathways may not result in overall health issues. An instant was reported in a paper that I found after submitting my question: http://www.ncbi.nlm.nih.gov/pubmed/10594757.
Thank you for your suggestions about using EdU/BrdU or Ki67 staining, and applying the reporter mouse strains! The question I proposed is kind of "weird" though: can we determine the sole contribution of cell migration to reepithelization, with the effect of proliferation ruled out or minimized. If this is achievable, we might compare the in vitro and in vivo migraton/re-epithelization studies on a better basis.
Might it be interesting and rational to test applying such an inhibitor like L-NIL used in the above paper?
I would rather be carefull with this type of drug, especially because there is no information about its effect on cell migration. It is also known that Nos is involved in the migration of endothelial cells (http://www.ncbi.nlm.nih.gov/pubmed/10323764), which would definitely impact the survival/migration/proliferation capacity of your cells of interest.
I fear that you will have to either find a really specific drug or to go for the reporter mice associated with a carefull analysis of the cell migration, proliferation, death and differentiation at different time points. Not the easiest, cheapest or quickest way, but that's how I would do it.
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