title: Store-operated CRAC channels regulate gene expression and proliferation in neural progenitor cells.
abstract: Calcium signals regulate many critical processes during vertebrate brain development including neurogenesis, neurotransmitter specification, and axonal outgrowth. However, the identity of the ion channels mediating Ca(2+) signaling in the developing nervous system is not well defined. Here, we report that embryonic and adult mouse neural stem/progenitor cells (NSCs/NPCs) exhibit store-operated Ca(2+) entry (SOCE) mediated by Ca(2+) release-activated Ca(2+) (CRAC) channels. SOCE in NPCs was blocked by the CRAC channel inhibitors La(3+), BTP2, and 2-APB and Western blots revealed the presence of the canonical CRAC channel proteins STIM1 and Orai1. Knock down of STIM1 or Orai1 significantly diminished SOCE in NPCs, and SOCE was lost in NPCs from transgenic mice lacking Orai1 or STIM1 and in knock-in mice expressing the loss-of-function Orai1 mutant, R93W. Therefore, STIM1 and Orai1 make essential contributions to SOCE in NPCs. SOCE in NPCs was activated by epidermal growth factor and acetylcholine, the latter occurring through muscarinic receptors. Activation of SOCE stimulated gene transcription through calcineurin/NFAT (nuclear factor of activated T cells) signaling through a mechanism consistent with local Ca(2+) signaling by Ca(2+) microdomains near CRAC channels. Importantly, suppression or deletion of STIM1 and Orai1 expression significantly attenuated proliferation of embryonic and adult NPCs cultured as neurospheres and, in vivo, in the subventricular zone of adult mice. These findings show that CRAC channels serve as a major route of Ca(2+) entry in NPCs and regulate key effector functions including gene expression and proliferation, indicating that CRAC channels are important regulators of mammalian neurogenesis.
authors.affiliation: Department of Molecular Pharmacology and Biological Chemistry, Feinberg School of Medicine, Northwestern University, Chicago, Illinois 60611.
authors.forename: Agila
authors.initials: A
authors.lastname: Somasundaram
authors.affiliation: Department of Molecular Pharmacology and Biological Chemistry, Feinberg School of Medicine, Northwestern University, Chicago, Illinois 60611.
authors.forename: Andrew K
authors.initials: AK
authors.lastname: Shum
authors.affiliation: Department of Discovery Toxicology, Amgen, Inc., Thousand Oaks, California 91320.
authors.forename: Helen J
authors.initials: HJ
authors.lastname: McBride
authors.affiliation: Department of Neurology, Northwestern University Feinberg School of Medicine, Northwestern University, Chicago, Illinois 60611, and.
authors.forename: John A
authors.initials: JA
authors.lastname: Kessler
authors.affiliation: Department of Pathology, New York University Langone Medical Center, New York, New York 10016.
authors.forename: Stefan
authors.initials: S
authors.lastname: Feske
authors.affiliation: Department of Molecular Pharmacology and Biological Chemistry, Feinberg School of Medicine, Northwestern University, Chicago, Illinois 60611.
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Miller
authors.affiliation: Department of Molecular Pharmacology and Biological Chemistry, Feinberg School of Medicine, Northwestern University, Chicago, Illinois 60611, m-prakriya@northwestern.edu.
authors.forename: Murali
authors.initials: M
authors.lastname: Prakriya
mesh_terms: D053687:Adult Stem Cells / Q000378:metabolism*; D000818:Animals; D017209:Apoptosis; D019703:Calcineurin / Q000502:physiology; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D020013:Calcium Signaling / Q000502:physiology*; D002455:Cell Division; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000378:metabolism*; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000502:physiology*; D017136:Ion Transport; D008562:Membrane Glycoproteins / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009116:Muscarine / Q000494:pharmacology; D050778:NFATC Transcription Factors / Q000378:metabolism; D058953:Neural Stem Cells / Q000378:metabolism*; D055495:Neurogenesis / Q000235:genetics / Q000502:physiology*; D000071740:ORAI1 Protein; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000494:pharmacology; D000071737:Stromal Interaction Molecule 1
chemical_list: D015220:Calcium Channels; D008562:Membrane Glycoproteins; D050778:NFATC Transcription Factors; D000071740:ORAI1 Protein; C520430:Orai1 protein, mouse; D034741:RNA, Small Interfering; C100757:Stim1 protein, mouse; D000071737:Stromal Interaction Molecule 1; D004815:Epidermal Growth Factor; D009116:Muscarine; D019703:Calcineurin; D002118:Calcium
keywords: CRAC channels; Orai1; STIM1; calcium; progenitor cell; proliferation