title: Chronic LiCl pretreatment suppresses thrombin-stimulated intracellular calcium mobilization through TRPC3 in astroglioma cells.
abstract: Transient receptor potential canonical type 3 (TRPC3) channels are activated in B lymphoblast cell lines from patients with bipolar disorder (BD), and its expression is reduced by chronic lithium treatment, implicating TRPC3 in the intracellular calcium (Ca2+) dyshomeostasis of BD. Thrombin, via a protease-activated receptor, moderates Ca2+signaling and TRPC3 in astrocytes, and also cell proliferation. We examined whether lithium pretreatment attenuates thrombin-stimulated TRPC3 expression and function in astrocytes, and levels of the calcium-binding peptide, S100B, which is expressed mainly in these cells.
  
  
  
  Human astroglioma, U-87MG, cells were pretreated with 1 mmol L-1LiCl for 1 day (acute), 3 days (subacute), and 7 days (chronic). To examine the role of TRPC3, genetically stable knockdown TRPC3 cells (TRPC3Lowcells) were constructed using U-87MG cells. Thrombin (2.0 U/mL)-stimulated Ca2+mobilization was measured by ratiometric fluorimetry. Changes in TRPC3 and S100B expression levels were determined by quantitative reverse transcription-polymerase chain reaction and immunoblotting, respectively. Cell proliferation was also measured using the WST-8 assay.
  
  
  
  In this cell model, thrombin-stimulated Ca2+mobilization, and both TRPC3 and S100B expression were suppressed by chronic LiCl pretreatment and the knockdown of TRPC3. Additionally, cell proliferation was attenuated in TRPC3Lowcells, compared with the negative control vector-transfected cell.
  
  
  
  The reduced Ca2+mobilization and S100B expression levels following chronic LiCl pretreatment and in TRPC3Lowcells support the notion that TRPC3 modulates S100B expression and is the target of the LiCl effect. Downregulation of TRPC3 may be an important mechanism by which lithium ameliorates pathophysiological intracellular Ca2+disturbances as observed in BD, accounting, in part, for its mood-stabilizing effects.
authors.affiliation: Laboratory of Cellular and Molecular Pathophysiology, Campbell Family Mental Health Research Institute, Centre for Addiction and Mental Health, Toronto, ON, Canada.|Department of Psychiatry, University of Toronto, Toronto, ON, Canada.|Department of Neuropsychiatry, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Chuo, Yamanashi, Japan.
authors.forename: Takuji
authors.initials: T
authors.lastname: Uemura
authors.affiliation: Laboratory of Cellular and Molecular Pathophysiology, Campbell Family Mental Health Research Institute, Centre for Addiction and Mental Health, Toronto, ON, Canada.
authors.forename: Marty
authors.initials: M
authors.lastname: Green
authors.affiliation: Laboratory of Cellular and Molecular Pathophysiology, Campbell Family Mental Health Research Institute, Centre for Addiction and Mental Health, Toronto, ON, Canada.|Department of Psychiatry, University of Toronto, Toronto, ON, Canada.|Department of Pharmacology and Toxicology, University of Toronto, Toronto, ON, Canada.|Institute of Medical Science, University of Toronto, Toronto, ON, Canada.|Program in Neuroscience, University of Toronto, Toronto, ON, Canada.
authors.forename: Jerry J
authors.initials: JJ
authors.lastname: Warsh
mesh_terms: D018692:Antimanic Agents / Q000494:pharmacology; D001254:Astrocytoma / Q000378:metabolism; D001714:Bipolar Disorder / Q000188:drug therapy / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D006801:Humans; D008094:Lithium / Q000494:pharmacology*; D064568:S100 Calcium Binding Protein beta Subunit / Q000378:metabolism*; D050052:TRPC Cation Channels / Q000378:metabolism*; D013917:Thrombin / Q000494:pharmacology
chemical_list: D018692:Antimanic Agents; D064568:S100 Calcium Binding Protein beta Subunit; C579160:S100B protein, human; D050052:TRPC Cation Channels; C104603:TRPC3 cation channel; D008094:Lithium; D013917:Thrombin; D002118:Calcium
keywords: S100B; TRPC3; intracellular calcium homeostasis; lithium; thrombin