Abstract:
:We investigated the early effects of the anti-idiotypic antibody (clone 1D5), which recognized the estrogen receptor (ER), on cytosolic free calcium concentration ([Ca2+]i) and its long term effects on creatine kinase (CK) specific activity in female human and rat osteoblasts. These actions were compared to the known membrane and genomic effects of 17 beta estradiol (E2). Like E2, clone 1D5 increased within 5 s [Ca2+]i in both cell types by two mechanisms: 1) Ca2+ influx through voltage-gated Ca2+ channels as shown by using EGTA a chelator of extracellular Ca2+, and nifedipine, a Ca2+ channel blocker; 2) Ca2+ mobilization from the endoplasmic reticulum as shown by using phospholipase C inhibitors, such as neomycin and U-73122, which involved a Pertussis toxin-sensitive G-protein. Clone 1D5 and E2 stimulated CK specific activity in human and rat osteoblasts with ten fold higher concentrations than those needed for the membrane effects (0.1 microgram/ml and 10 pM, respectively). Both effects were gender-specific since testosterone and 5 alpha-dihydotesterone were uneffective. Tamoxifen and Raloxifene, two estrogen nuclear antagonists, inhibited CK response to 1D5 and E2 and Ca2+ response to 1D5, but not Ca2+ response to E2. By contrast, (Fab')2 dimer, a proteolytic fragment of 1D5 with antagonist properties, inhibited both membrane and genomic effects of 1D5 and E2. In conclusion, these results imply that clone 1D5 has an estrogen like activity both at the membrane and nuclear levels in female human and rat osteoblasts. 1D5 must therefore interact with membrane binding sites, penetrate the cells, and reach the nuclear receptors by an as yet uncharacterized mechanism.
journal_name
J Cell Biochemjournal_title
Journal of cellular biochemistryauthors
Sömjen D,Kohen F,Lieberherr Mdoi
10.1002/(sici)1097-4644(199704)65:1<53::aid-jcb6>3subject
Has Abstractpub_date
1997-04-01 00:00:00pages
53-66issue
1eissn
0730-2312issn
1097-4644pii
10.1002/(SICI)1097-4644(199704)65:1<53::AID-JCB6>3journal_volume
65pub_type
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