Analysis Tools
nervous system
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• apical shift in migrating CPNs is less pronounced when CPNs are labeled with EdU at E16
• EdU birthdating revealed impaired migration of upper layer (UL) cortical projection neurons (CPNs) into the cortical plate
• CPNs labeled with EdU at E14 show an apical shift of ~100 um at E18.5; a larger proportion of E14-born UL CPNs remain restricted to the deeper cortex bins
• however, no defects in the formation of Reelin+ CR cells nor in radial glial scaffolding are observed
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• neurofilament (NF+) fiber staining showed a significant decrease in the thickness of cortical white matter tracts at E16.5 and E18.5
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• neurofilament (NF+) fiber staining is significantly reduced in the corpus callosum at E18.5, reflecting a severe reduction in crossing callosal fibers
• DiI labeling of axonal projections is undetectable in the corpus callosum at E18.5, indicating impaired development of callosal projections
• in contrast, corticothalamic projections of the internal capsule remain unaffected
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• coronal sections of the corpus callosum labeled with acetylated alpha-tubulin and Tubb3 showed decreased corpus callosum thickness at E18.5
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• formation of UL CPNs is perturbed with progressively decreased numbers of Satb2+ cells in the upper bins from E16.5 to E18.5
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• at E18.5, an increased proportion of DAPI+ nuclei are localized apically, towards the ventricular zone (VZ), relative to controls
• fewer Tbr2+ intermediate (basal) progenitors, which give rise to UL 2/3 CPNs, are found within deep regions at E18.5, suggesting altered formation and/or migration of nascent neurons from the subventricular zone (SVZ)
• marker analysis of UL CPNs showed decreased Satb2+ cell numbers and an apical shift of Satb2 expression starting at E16.5 and increasing in severity by E18.5; similarly, CDP/Cux1+ cell numbers are normal at E14.5 but start to decrease at E16.5 and are severely reduced at E18.5
• analysis of deep layer (DL) markers showed an apical shift in the expression domains of both Tbr1 (DL6-specific) and Ctip2 (DL5-specific) from E16.5 to E18.5, consistent with reduced cortical thickness and impaired UL formation
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• at E18.5, thinning of the cortex is noted in the cortical plate and white matter below it
• cortex thinning is first noted at E16.5, increasing in severity by E18.5
• no thinning is seen at E14.5 when UL CPNs are born
• no increase in apoptosis is noted in cortical slices at E14.5 or E18.5
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• at P28, Golgi-stained cortices show a severe reduction in total neurite outgrowth (length) and decreased branching complexity in mature UL2/3 CPNs in the cortex
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• at P28, Golgi-stained UL CPNs show greatly attenuated dendritic arborization morphologies
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• at P28, Golgi-stained UL2/3 CPNs show reduced dendritic branching complexity in the cortex; overall mature CPN morphology is much less complex with less elaborately branched neurites
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cellular
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• apical shift in migrating CPNs is less pronounced when CPNs are labeled with EdU at E16
• EdU birthdating revealed impaired migration of upper layer (UL) cortical projection neurons (CPNs) into the cortical plate
• CPNs labeled with EdU at E14 show an apical shift of ~100 um at E18.5; a larger proportion of E14-born UL CPNs remain restricted to the deeper cortex bins
• however, no defects in the formation of Reelin+ CR cells nor in radial glial scaffolding are observed
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