title: Endothelin1-induced Ca(2+) mobilization is altered in calvarial osteoblastic cells of Cx43(+/- ) mice.
abstract: During bone remodeling, osteoblastic (OB) cells have a central role leading to the production of extracellular matrix and its subsequent mineralization. As revealed by human physiopathologies, the OB differentiation process is essential for the control of calcium metabolism and normal bone formation. Moreover, accumulating data in the field of bone development suggest that connexin 43 (Cx43)-mediated gap junctional communication plays an important role in OB differentiation and function. Since Ca(2+) has a central role in OB physiology, the aim of the present study was to investigate the hypothetical involvement of Cx43 in OB calcium homeostasis. We performed measurements of intracellular calcium activity ([Ca(2+)]( i )) by a cytofluorimetric method using Fluo-4 as a calcium indicator and endothelin-1 (ET-1) as a physiological calcium-mobilizing factor on cultured OB cells isolated from calvaria of Cx43(+/-) and Cx43(+/+) mice. Partial deletion of the Cx43 gene induced a significant decrease in the [Ca(2+)]( i ) rise elicited by ET-1. This reduction was not correlated to a decrease or a modification of ET receptor subtype expression as assessed by real-time reverse-transcription polymerase chain reaction. Pharmacological investigations led us to demonstrate that the significant difference in [Ca(2+)]( i ) peak amplitude during the ET-1 action was associated with decreased calcium influx involving L-type voltage-sensitive calcium channels, whereas calcium release from intracellular stores and implication of phospholipase C were not affected by the reduced expression of Cx43. In conclusion, our data demonstrate for the first time that the Cx43 level of expression and/or function is able to modulate the [Ca(2+)]( i ) mobilization in OB cells.
authors.affiliation: Institut de Physiologie et de Biologie Cellulaires, CNRS UMR6187, Université de Poitiers, 40 avenue du recteur Pineau, 86022, Poitiers, France.
authors.forename: Graziello
authors.initials: G
authors.lastname: Geneau
authors.forename: Norah
authors.initials: N
authors.lastname: Defamie
authors.forename: Marc
authors.initials: M
authors.lastname: Mesnil
authors.forename: Laurent
authors.initials: L
authors.lastname: Cronier
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling; D002450:Cell Communication; D002478:Cells, Cultured; D018031:Connexin 43 / Q000235:genetics / Q000502:physiology*; D019332:Endothelin-1 / Q000494:pharmacology*; D017629:Gap Junctions / Q000378:metabolism; D006579:Heterozygote; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D010006:Osteoblasts / Q000166:cytology / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D044022:Receptor, Endothelin A / Q000378:metabolism; D044023:Receptor, Endothelin B / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012886:Skull / Q000166:cytology / Q000378:metabolism*; D010738:Type C Phospholipases / Q000378:metabolism
chemical_list: D018031:Connexin 43; D019332:Endothelin-1; D012333:RNA, Messenger; D044022:Receptor, Endothelin A; D044023:Receptor, Endothelin B; D010738:Type C Phospholipases; D002118:Calcium