Defective extracellular calcium (Ca(o))-sensing receptor (CaR)-mediated stimulation of a Ca(2+)-activated potassium channel in glioblastoma cells transfected with a dominant negative CaR.
Glioblastoma cells exhibit several forms of sensitivity to extracellular calcium (Ca(o)) that might be conferred by the Ca(o)-sensing receptor (CaR) that is intimately involved in the maintenance of Ca(o) homeostasis by various cell types. This receptor is expressed in human glioblastoma cell line, U87, and here we show that CaR activators stimulate a Ca(2+)-activated potassium (K(+)) channel (CAKC) with a conductance of 140 pS. The responses to CaR activators, however, were blunted in U87 cells transfected with a CaR bearing an inactivating mutation (R185Q) that has previously been shown to exert a dominant negative (DN) action on the wild type receptor. Raising Ca(o) from 0.75 to 2.0 mM or addition of a polycationic CaR agonist, each activated CAKC in nontransfected wild type and empty vector-transfected U87 cells, while they had little or no effect on channel activity in cells expressing the DN CaR (DN-CaR cells). In nontransfected wild type and empty vector-transfected cells, the specific 'calcimimetic' CaR activator, NPS R-467, stimulated the channel, while its less active stereoisomer, NPS S-467, did not. In DN-CaR cells, in contrast, NPS R-467, had no effect on channel activity, suggesting defective coupling of the CaR to this ion channel. CaR-mediated stimulation of these K(+) channels could lead to membrane repolarization and related changes in cellular function under normal conditions. Since the R185Q mutation in the CaR produces a more severe phenotype in humans than most inactivating mutations of this receptor, some of its clinical consequences could potentially result from abnormal CaR-dependent channel functioning.
Ye
C
C
Endocrine-Hypertension Division and Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, 221 Longwood Avenue, 02115, Boston, MA, USA.
Chattopadhyay
N
N
Brown
E M
EM
Vassilev
P M
PM
D002118:Calcium / Q000378:metabolism / Q000494:pharmacology*; D005110:Extracellular Space / Q000737:chemistry / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D005909:Glioblastoma; D006706:Homeostasis / Q000502:physiology; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D016296:Mutagenesis / Q000502:physiology; D018408:Patch-Clamp Techniques; D011188:Potassium / Q000378:metabolism; D015221:Potassium Channels / Q000378:metabolism*; D044169:Receptors, Calcium-Sensing; D011956:Receptors, Cell Surface / Q000235:genetics* / Q000378:metabolism*; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000737:chemistry / Q000378:metabolism
D015221:Potassium Channels; D044169:Receptors, Calcium-Sensing; D011956:Receptors, Cell Surface; D011188:Potassium; D002118:Calcium