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Birthdating cells

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Furthermore, Lpar1-GFP and WT pups from similar overnight matings but without Brd U injections were transcardially perfused with 4% PFA and postfixed in 4% PFA overnight for marker colocalization analysis.For temporal GFP expression analysis, additionally, adult Lpar1-GFP males were obtained.Subplate cells in the mouse are generally defined as cells located in the subplate layer between the white matter and layer 6a.They are some of the earliest born and maturing cells of the cerebral cortex. All of the early birthdating studies on rodent preplate and subplate cells were of rats. (1977) and Bayer and Altman (1990) determined that rat preplate neurons are generated from E14 (plug date = E1).

Alternatively, immunohistochemistry was performed against the pan-neuronal marker Neu N and Brd U.

We replicated some of these findings, extending them to a general loss of early-born neurons during the first postnatal week, and determined whether there is a differential survival among marker-labeled neurons of the mouse subplate.

For the purposes of this study, the subplate zone or layer was defined as a 50 µm thick band directly above the WM (Hoerder-Suabedissen and Molnár 2012) and is equivalent to layer 6b. All 4 molecular markers used here label a thin band of cells in the subplate location with very few cells above (in layer 6a) or below (in WM).

None of these, however, is expressed from the time of subplate cell generation.

Thus, we set out to determine the birthdates of the marker-labeled subplate neurons.