Role of L-type Ca2+ channels in neural stem/progenitor cell differentiation

Eur J Neurosci. 2006 Feb;23(4):935-44. doi: 10.1111/j.1460-9568.2006.04628.x.


Ca(2+) influx through voltage-gated Ca(2+) channels, especially the L-type (Ca(v)1), activates downstream signaling to the nucleus that affects gene expression and, consequently, cell fate. We hypothesized that these Ca(2+) signals may also influence the neuronal differentiation of neural stem/progenitor cells (NSCs) derived from the brain cortex of postnatal mice. We first studied Ca(2+) transients induced by membrane depolarization in Fluo 4-AM-loaded NSCs using confocal microscopy. Undifferentiated cells (nestin(+)) exhibited no detectable Ca(2+) signals whereas, during 12 days of fetal bovine serum-induced differentiation, neurons (beta-III-tubulin(+)/MAP2(+)) displayed time-dependent increases in intracellular Ca(2+) transients, with DeltaF/F ratios ranging from 0.4 on day 3 to 3.3 on day 12. Patch-clamp experiments revealed similar correlation between NSC differentiation and macroscopic Ba(2+) current density. These currents were markedly reduced (-77%) by Ca(v)1 channel blockade with 5 microm nifedipine. To determine the influence of Ca(v)1-mediated Ca(2+) influx on NSC differentiation, cells were cultured in differentiative medium with either nifedipine (5 microm) or the L-channel activator Bay K 8644 (10 microm). The latter treatment significantly increased the percentage of beta-III-tubulin(+)/MAP2(+) cells whereas nifedipine produced opposite effects. Pretreatment with nifedipine also inhibited the functional maturation of neurons, which responded to membrane depolarization with weak Ca(2+) signals. Conversely, Bay K 8644 pretreatment significantly enhanced the percentage of responsive cells and the amplitudes of Ca(2+) transients. These data suggest that NSC differentiation is strongly correlated with the expression of voltage-gated Ca(2+) channels, especially the Ca(v)1, and that Ca(2+) influx through these channels plays a key role in promoting neuronal differentiation.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aniline Compounds / metabolism
  • Animals
  • Animals, Newborn
  • Calcium Channel Blockers / pharmacology
  • Calcium Channels, L-Type / physiology*
  • Calcium Signaling / physiology
  • Cell Count / methods
  • Cell Differentiation / drug effects
  • Cell Differentiation / physiology*
  • Cells, Cultured
  • Cerebral Cortex / cytology*
  • Glial Fibrillary Acidic Protein / metabolism
  • Immunohistochemistry / methods
  • Intermediate Filament Proteins / metabolism
  • Membrane Potentials / drug effects
  • Membrane Potentials / physiology
  • Membrane Potentials / radiation effects
  • Mice
  • Microscopy, Confocal / methods
  • Microtubule-Associated Proteins / metabolism
  • Nerve Tissue Proteins / metabolism
  • Nestin
  • Neurons / cytology*
  • Nifedipine / pharmacology
  • Patch-Clamp Techniques / methods
  • Potassium Chloride / pharmacology
  • Protein Serine-Threonine Kinases / metabolism
  • Receptor-Interacting Protein Serine-Threonine Kinases
  • Sodium Channel Blockers / pharmacology
  • Stem Cells / physiology*
  • Tetrodotoxin / pharmacology
  • Time Factors
  • Tubulin / metabolism
  • Tumor Necrosis Factor Receptor-Associated Peptides and Proteins / metabolism
  • Xanthenes / metabolism


  • Aniline Compounds
  • Calcium Channel Blockers
  • Calcium Channels, L-Type
  • Fluo 4
  • Glial Fibrillary Acidic Protein
  • Intermediate Filament Proteins
  • Microtubule-Associated Proteins
  • Mtap2 protein, mouse
  • NES protein, human
  • Nerve Tissue Proteins
  • Nes protein, mouse
  • Nestin
  • Sodium Channel Blockers
  • Tubulin
  • Tumor Necrosis Factor Receptor-Associated Peptides and Proteins
  • Xanthenes
  • Tetrodotoxin
  • Potassium Chloride
  • Protein Serine-Threonine Kinases
  • Receptor-Interacting Protein Serine-Threonine Kinases
  • Ripk1 protein, mouse
  • Nifedipine