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Journal of Neuroscience, Vol 15, 5014-5024, Copyright © 1995 by Society for Neuroscience
Persistence of early-generated neurons in the rodent subplate: assessment of cell death in neocortex during the early postnatal period
F Valverde, L Lopez-Mascaraque, M Santacana and JA De Carlos
Laboratorio de Neuroanatomia Comparada, Instituto Cajal (CSIC), Madrid, Spain.
In the rat, the deepest neocortical layer forms a conspicuous cell band
known as layer Vlb. Cells in layer Vlb are among the first to
differentiate, and it has been regarded as an homolog to the subplate of
primates and carnivores. Cell death has been considered a universal feature
of subplate cells. In order to assess the validity of this assertion, we
examined the sequence of generation and the extent of cell death in layer
Vlb. This was achieved using injections of 3H- thymidine and two methods
for the direct visualization of apoptotic figures. Single injections of
3H-thymidine were performed between E12 and E15 (E0 is the day of
insemination), and brains were examined at different postnatal ages between
P1 and P63. The number of heavily labeled cells were counted in layer Vlb
in six standard, equally spaced coronal sections in each brain. Single
injections at E12 labels about 3% of the entire population of layer Vlb
cells, 17% at E13, 30% at E14, and < 1% at E15. Our results indicate
that the absolute number of heavily labeled cells in layer Vlb remains
constant. The analysis of variance (one-way ANOVA) showed that the
difference among the group means was not significant from P1 to P63 after
injections at either E12, E13, or E14. In order to confirm these results,
we evaluated the distribution of pyknotic (apoptotic) cell bodies in the
neocortex. Apoptotic cells were visualized in Nissl preparations and by
histochemical staining using an in situ apoptosis detection kit. The
analysis was performed in rats from E18 to P15. Both methods gave
comparable results. We found that the amount of cell death in layer Vlb is
neither particularly prominent nor significantly different from that which
occurs in the remaining neocortical layers, apart from layer II and in the
white matter of the corpus callosum. We conclude that neuronal death does
not play any significant role in the rodent subplate.
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