Cerebral Cortex, Vol. 10, No. 6, 561-573,
June 2000
© 2000 Oxford University Press
Developmental Changes in Cell Calcium Homeostasis during Neurogenesis of the Embryonic Rat Cerebral Cortex
Laboratory of Neurophysiology, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892, USA
We quantified cytoplasmic Ca2+ (Ca2+c) levels in cells dissociated from the embryonic (E) rat cortex during neurogenesis. Dualrecordings by flow cytometry using calcium and voltage-sensitive dyes revealed that, at the beginning of cortical development (E11–12), precursor cells exhibited either low (<100 nM), moderate (~250 nM) or high (>1 µM) resting Ca2+c levels and well-polarized (–70 mV) or less-polarized (–40 mV) resting membrane potentials which reflected postmitotic or proliferative stages of the cell cycle. Ca2+c levels of all cells included a Ca2+o entry component, which was also Mn2+-permeant in actively proliferating precursors. Postmitotic, but not premitotic, precursors exhibited thapsigargin-sensitive intracellular Ca2+ (Ca2+i) stores, which had similar capacities throughout neuronal lineage development. Differentiating neurons, but not precursors expressed Ca2+i stores with ryanodine and caffeine sensitivity and baseline Ca2+c levels that depended on Na+–Ca2+ exchange activity. Voltage-dependent Ca2+o entry was not detected in precursors, but emerged during neuronal differentiation, with most of the neurons expressing functional L-type Ca2+ channels. Ca2+ imaging of individually immunoidentified cells acutely recovered in culture confirmed that precursors differentiate into neurons which stereotypically exhibit Ca2+o entry at the level of the membrane with increased Ca2+i release mechanisms on Ca2+i stores, Na+–Ca2+ exchange activity and expression of voltagedependent Ca2+ channels.
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