title: Store-operated calcium entry is essential for glial calcium signalling in CNS white matter.
abstract: 'Calcium signalling' is the ubiquitous response of glial cells to multiple extracellular stimuli. The primary mechanism of glial calcium signalling is by release of calcium from intracellular stores of the endoplasmic reticulum (ER). Replenishment of ER Ca2+stores relies on store-operated calcium entry (SOCE). However, despite the importance of calcium signalling in glial cells, little is known about their mechanisms of SOCE. Here, we investigated SOCE in glia of the mouse optic nerve, a typical CNS white matter tract that comprises bundles of myelinated axons and the oligodendrocytes and astrocytes that support them. Using quantitative RT-PCR, we identified Orai1 channels, both Stim1 and Stim2, and the transient receptor potential M3 channel (TRPM3) as the primary channels for SOCE in the optic nerve, and their expression in both astrocytes and oligodendrocytes was demonstrated by immunolabelling of optic nerve sections and cultures. The functional importance of SOCE was demonstrated by fluo-4 calcium imaging on isolated intact optic nerves and optic nerve cultures. Removal of extracellular calcium ([Ca2+]o) resulted in a marked depletion of glial cytosolic calcium ([Ca2+]i), which recovered rapidly on restoration of [Ca2+]ovia SOCE. 2-aminoethoxydiphenylborane (2APB) significantly decreased SOCE and severely attenuated ATP-mediated calcium signalling. The results provide evidence that Orai/Stim and TRPM3 are important components of the 'calcium toolkit' that underpins SOCE and the sustainability of calcium signalling in white matter glia.
authors.affiliation: Institute of Biomedical and Biomolecular Sciences, School of Pharmacy and Biomedical Science, University of Portsmouth, Portsmouth, PO1 2DT, UK.
authors.forename: M
authors.initials: M
authors.lastname: Papanikolaou
authors.affiliation: Institute of Biomedical and Biomolecular Sciences, School of Pharmacy and Biomedical Science, University of Portsmouth, Portsmouth, PO1 2DT, UK.
authors.forename: A
authors.initials: A
authors.lastname: Lewis
authors.affiliation: Institute of Biomedical and Biomolecular Sciences, School of Pharmacy and Biomedical Science, University of Portsmouth, Portsmouth, PO1 2DT, UK. arthur.butt@port.ac.uk.
authors.forename: A M
authors.initials: AM
authors.lastname: Butt
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001896:Boron Compounds / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling / Q000502:physiology*; D018507:Gene Expression Regulation, Developmental / Q000235:genetics*; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D018991:Myelin Proteolipid Protein / Q000235:genetics / Q000378:metabolism; D009457:Neuroglia / Q000378:metabolism* / Q000648:ultrastructure; D000071740:ORAI1 Protein / Q000235:genetics / Q000378:metabolism; D009900:Optic Nerve / Q000166:cytology*; D009924:Organ Culture Techniques; D012333:RNA, Messenger / Q000378:metabolism; D000071737:Stromal Interaction Molecule 1 / Q000235:genetics / Q000378:metabolism; D000071738:Stromal Interaction Molecule 2 / Q000235:genetics / Q000378:metabolism; D050053:TRPM Cation Channels / Q000235:genetics / Q000378:metabolism; D066127:White Matter / Q000378:metabolism*
chemical_list: C504876:2-aminoethoxydiphenylborane; D001896:Boron Compounds; D005904:Glial Fibrillary Acidic Protein; D008164:Luminescent Proteins; D018991:Myelin Proteolipid Protein; D000071740:ORAI1 Protein; C520430:Orai1 protein, mouse; D012333:RNA, Messenger; C100757:Stim1 protein, mouse; C525024:Stim2 protein, mouse; D000071737:Stromal Interaction Molecule 1; D000071738:Stromal Interaction Molecule 2; D050053:TRPM Cation Channels; C533868:TRPM3 protein, mouse; C413663:fluorescent protein 583; D002118:Calcium
keywords: Astrocyte; CRAC; Calcium signalling; Glia; Oligodendrocyte; Store-operated calcium channel; TRP channel; White matter