Extremely low-frequency electromagnetic fields promote in vitro neurogenesis via upregulation of Ca(v)1-channel activity.
We previously reported that exposure to extremely low-frequency electromagnetic fields (ELFEFs) increases the expression and function of voltage-gated Ca2+)channels and that Ca2+ influx through Ca(v)1 channels plays a key role in promoting the neuronal differentiation of neural stem/progenitor cells (NSCs). The present study was conducted to determine whether ELFEFs influence the neuronal differentiation of NSCs isolated from the brain cortices of newborn mice by modulating Ca(v)1-channel function. In cultures of differentiating NSCs exposed to ELFEFs (1 mT, 50 Hz), the percentage of cells displaying immunoreactivity for neuronal markers (beta-III-tubulin, MAP2) and for Ca(v)1.2 and Ca(v)1.3 channels was markedly increased. NSC-differentiated neurons in ELFEF-exposed cultures also exhibited significant increases in spontaneous firing, in the percentage of cells exhibiting Ca2+ transients in response to KCl stimulation, in the amplitude of these transients and of Ca2+ currents generated by the activation of Ca(v)1 channels. When the Ca(v)1-channel blocker nifedipine (5 microM) was added to the culture medium, the neuronal yield of NSC differentiation dropped significantly, and ELFEF exposure no longer produced significant increases in beta-III-tubulin- and MAP2-immunoreactivity rates. In contrast, the effects of ELFEFs were preserved when NSCs were cultured in the presence of either glutamate receptor antagonists or Ca(v)2.1- and Ca(v)2.2-channel blockers. ELFEF stimulation during the first 24 h of differentiation caused Ca(v)1-dependent increases in the number of cells displaying CREB phosphorylation. Our data suggest that ELFEF exposure promotes neuronal differentiation of NSCs by upregulating Ca(v)1-channel expression and function.
Piacentini
Roberto
R
Institute of Human Physiology, Medical School, Catholic University S. Cuore, Rome, Italy.
Ripoli
Cristian
C
Mezzogori
Daniele
D
Azzena
Gian Battista
GB
Grassi
Claudio
C
D000818:Animals; D020746:Calcium Channels, L-Type / Q000378:metabolism*; D002454:Cell Differentiation; D049109:Cell Proliferation; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism; D004574:Electromagnetic Fields; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009474:Neurons / Q000166:cytology*; D010641:Phenotype; D010766:Phosphorylation; D013234:Stem Cells / Q000166:cytology; D013997:Time Factors; D014404:Tubulin / Q000378:metabolism; D015854:Up-Regulation
C514350:CACNA1C protein, mouse; C485995:Cacna1d protein, mouse; D020746:Calcium Channels, L-Type; D017362:Cyclic AMP Response Element-Binding Protein; D008869:Microtubule-Associated Proteins; C079240:Mtap2 protein, mouse; D014404:Tubulin