Expression of mRNAs for the alpha 1 subunit of voltage-gated calcium channels in human osteoblast-like cell lines and in normal human osteoblasts.
The activation of osteoblast calcium channels by many bone regulatory factors suggests an important role for intracellular calcium signaling in the control of bone remodeling. At least six different genes for the alpha 1 subunit of voltage-gated calcium channels have been cloned including L-type (alpha 1S, alpha 1C, and alpha 1D) and non-L-type (alpha 1A, alpha 1B, and alpha 1E) isoforms. The goal of the present study was to identify which of these calcium channel isoforms are transcribed in human osteoblast-like cell lines (hFOB, MG-63, SAOS-2, TE-85, G-292) and in cultures of normal human osteoblasts. Reverse transcriptase-PCR was used to amplify sequences corresponding to each of the alpha 1 subunits using isoform specific primers. The products of the PCR reaction were cloned and sequenced to verify their identify and used to probe southern blots of the PCR reactions. The results indicate that among the different types of osteoblast-like cells examined, two calcium channel isoforms were always expressed (alpha 1C and alpha 1A), three isoforms were variably expressed (alpha 1S, alpha 1D and alpha 1B), and one isoform was not expressed in any of the osteoblast-like cells (alpha 1E) but was easily detected in human brain controls. Our results indicate that mRNAs for multiple calcium channel alpha 1 subunits are expressed in human osteoblasts, including both L-type and non-L-type isoforms. In addition, significant heterogeneity exists between the different osteoblast cell models examined in the type and mRNA abundance of the different calcium channel isoforms.
Barry
E L
EL
Department of Pharmacology and Toxicology, Dartmouth Medical School, Hanover, New Hampshire 03755-3835, USA.
D001859:Bone Neoplasms; D015220:Calcium Channels / Q000235:genetics*; D020746:Calcium Channels, L-Type / Q000235:genetics; D002460:Cell Line; D017931:DNA Primers; D006801:Humans; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism*; D012516:Osteosarcoma; D020033:Protein Isoforms / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D015220:Calcium Channels; D020746:Calcium Channels, L-Type; D017931:DNA Primers; D020033:Protein Isoforms; D012333:RNA, Messenger