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Calcium control of gene regulation in rat hippocampal neuronal cultures
Author(s) -
Pinato Giulietta,
Pegoraro Silvia,
Iacono Giovanni,
Ruaro Maria Elisabetta,
Torre Vincent
Publication year - 2009
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.21820
Subject(s) - nmda receptor , voltage dependent calcium channel , hippocampal formation , microbiology and biotechnology , chemistry , calcium in biology , receptor , patch clamp , calcium , biology , neuroscience , intracellular , biochemistry , organic chemistry
Abstract Blockage of GABA‐A receptors in hippocampal neuronal cultures triggers synchronous bursts of spikes initiating neuronal plasticity, partly mediated by changes of gene expression. By using specific pharmacological blockers, we have investigated which sources of Ca 2+ entry primarily control changes of gene expression induced by 20 µM gabazine applied for 30 min (GabT). Intracellular Ca 2+ transients were monitored with Ca 2+ imaging while recording electrical activity with patch clamp microelectrodes. Concomitant transcription profiles were obtained using Affymetrix oligonucleotide microarrays and confirmed with quantitative RT‐PCR. Blockage of NMDA receptors with 2‐amino‐5‐phosphonovaleric acid (APV) did not reduce significantly somatic Ca 2+ transients, which, on the contrary, were reduced by selective blockage of L, N, and P/Q types voltage gated calcium channels (VGCCs). Therefore, we investigated changes of gene expression in the presence of blockers of NMDA receptors and L, N, and P/Q VGCCs. Our results show that: (i) among genes upregulated by GabT, there are genes selectively dependent on NMDA activation, genes selectively dependent on L‐type VGCCs and genes dependent on the activation of both channels; (ii) the majority of genes requires the concomitant activation of NMDA receptors and Ca 2+ entry through VGCCs; (iii) blockage of N and P/Q VGCCs has an effect similar but not identical to blockage of L‐type VGCCs. J. Cell. Physiol. 220: 727–747, 2009. © 2009 Wiley‐Liss, Inc.