Abstract
DNA complementary to bovine preproparathyroid hormone mRNA was cloned and labeled by nick translation in order to measure mRNA by molecular hybridization. Bovine parathyroid cells were maintained in primary tissue culture for periods up to 96 h at 0.5 mM, 1.25 mM, and 2.5 mM calcium, which was followed by extraction of cellular RNA. Levels of mRNA showed no differences at 0.5 or 1.25 mM calcium, but at high calcium levels, there was a reversible decrease that began at 16 h to a plateau at 30% of control after 72 h. These studies suggest that the glandular capacity to synthesize hormone may be at or near maximal at normal calcium, but at high calcium, there is a decrease over time in steady state levels of mRNA.
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- Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brown E. M., Hurwitz S., Aurbach G. D. Preparation of viable isolated bovine parathyroid cells. Endocrinology. 1976 Dec;99(6):1582–1588. doi: 10.1210/endo-99-6-1582. [DOI] [PubMed] [Google Scholar]
- Habener J. F., Kemper B., Potts J. T., Jr Calcium-dependent intracellular degradation of parathyroid hormone: a possible mechanism for the regulation of hormone stores. Endocrinology. 1975 Aug;97(2):431–441. doi: 10.1210/endo-97-2-431. [DOI] [PubMed] [Google Scholar]
- Hanley D. A., Takatsuki K., Birnbaumer M. E., Schneider A. B., Sherwood L. M. In vitro perifusion for the study of parathyroid hormone secretion: effects of extracellular calcium concentration and beta-adrenergic regulation on bovine parathyroid hormone secretion in vitro. Calcif Tissue Int. 1980;32(1):19–27. doi: 10.1007/BF02408518. [DOI] [PubMed] [Google Scholar]
- Hanley D. A., Takatsuki K., Sultan J. M., Schneider A. B., Sherwood L. M. Direct release of parathyroid hormone fragments from functioning bovine parathyroid glands in vitro. J Clin Invest. 1978 Dec;62(6):1247–1254. doi: 10.1172/JCI109245. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Heinrich G., Kronenberg H. M., Potts J. T., Jr, Habener J. F. Parathyroid hormone messenger ribonucleic acid: effects of calcium on cellular regulation in vitro. Endocrinology. 1983 Feb;112(2):449–458. doi: 10.1210/endo-112-2-449. [DOI] [PubMed] [Google Scholar]
- Kronenberg H. M., McDevitt B. E., Majzoub J. A., Nathans J., Sharp P. A., Potts J. T., Jr, Rich A. Cloning and nucleotide sequence of DNA coding for bovine preproparathyroid hormone. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4981–4985. doi: 10.1073/pnas.76.10.4981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Merril C. R., Goldman D., Sedman S. A., Ebert M. H. Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science. 1981 Mar 27;211(4489):1437–1438. doi: 10.1126/science.6162199. [DOI] [PubMed] [Google Scholar]
- Moran J. R., Born W., Tuchschmid C. R., Fischer J. A. Calcium-regulated biosynthesis of the parathyroid secretory protein, proparathyroid hormone, and parathyroid hormone in dispersed bovine parathyroid cells. Endocrinology. 1981 Jun;108(6):2264–2268. doi: 10.1210/endo-108-6-2264. [DOI] [PubMed] [Google Scholar]
- Ricciardi R. P., Miller J. S., Roberts B. E. Purification and mapping of specific mRNAs by hybridization-selection and cell-free translation. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4927–4931. doi: 10.1073/pnas.76.10.4927. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sherwood L. M., Burstein Y., Schechter I. Primary structure of the NH2-terminal extra piece of the precursor to human placental lactogen. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3819–3823. doi: 10.1073/pnas.76.8.3819. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Strair R. K., Yap S. H., Shafritz D. A. Use of molecular hybridization to purify and analyze albumin messenger RNA from rat liver. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4346–4350. doi: 10.1073/pnas.74.10.4346. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weaver C. A., Gordon D. F., Kemper B. Nucleotide sequence of bovine parathyroid hormone messenger RNA. Mol Cell Endocrinol. 1982 Nov-Dec;28(3):411–424. doi: 10.1016/0303-7207(82)90136-8. [DOI] [PubMed] [Google Scholar]
- White B. A., Bancroft F. C. Cytoplasmic dot hybridization. Simple analysis of relative mRNA levels in multiple small cell or tissue samples. J Biol Chem. 1982 Aug 10;257(15):8569–8572. [PubMed] [Google Scholar]


