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Injection of bovine parathyroid poly(A) + RNA into xenopus oocytes confers sensitivity to high extracellular calcium
Author(s) -
Chen TsuiHua,
Pratt Stacy A.,
Shoback Dolores M.
Publication year - 1994
Publication title -
journal of bone and mineral research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.882
H-Index - 241
eISSN - 1523-4681
pISSN - 0884-0431
DOI - 10.1002/jbmr.5650090219
Subject(s) - extracellular , parathyroid hormone , xenopus , rna , messenger rna , calcium , intracellular , chemistry , endocrinology , medicine , microbiology and biotechnology , efflux , biology , biochemistry , gene
Parathyroid cells detect increments in the extracellular [Ca 2+ ], which lead to substantial increases in intracellular free Ca 2+ ([Ca 2+ ] i ) and, ultimately, to suppression of parathyroid hormone (PTH) secretion. To determine whether mRNA from parathyroid tissue could confer sensitivity to high extracellular Ca 2+ , we isolated and injected total bovine parathyroid poly(A) + RNA into Xenopus laevis oocytes. To assess translational activity of the RNA, PTH released into the media was measured. Intact PTH was detected in the medium for ≤48 h, and injection of increasing amounts of RNA (∼0.5–50 ng/oocyte) led to the release of greater quantities of PTH. We screened for the expression of a putative Ca 2+ sensor molecule by measuring 45 Ca efflux from preloaded oocytes, in response to raising extracellular [Ca 2+ ] from 0.7 to 5.7 mM. This increment in [Ca 2+ ] stimulated 45 Ca efflux by 249 ± 52 cpm over 20 min from eggs injected with parathyroid poly(A) + RNA ( n = 22). This response was significantly greater than 45 Ca efflux from any group of controls exposed to the same change in extracellular Ca 2+ ( p < 0.02), including oocytes injected with either water, cRNA for the platelet‐derived growth factor (PDGF) BB receptor, or T cell poly(A) + RNA. Size‐fractionation of poly(A) + RNA over sucrose gradients demonstrated that mRNA, which induced responsiveness to high extracellular Ca 2+ , was present in fractions with transcripts of ˜5–9 kB. Injection of these fractions also conferred sensitivity to the presence of Ba 2+ or Sr 2+ (both at 5 mM) in the media. These findings establish that parathyroid cells express mRNA for a molecule capable of detecting changes in the extracellular divalent cation concentration. This molecule may play a role in secretory responses mediated by Ca 2+ in the parathyroid.