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Endocrinology, Vol 132, 2090-2098, Copyright © 1993 by Endocrine Society
ARTICLES |
FR Bringhurst, H Juppner, J Guo, P Urena, JT Potts Jr, HM Kronenberg, AB Abou- Samra and GV Segre
Endocrine Unit, Massachusetts General Hospital, Boston 02114.
PTH elicits multiple second messenger signals in target cells. This signaling diversity may reflect coupling of a single species of PTH receptors to multiple effectors, the action of different subtypes of PTH receptors, or both. We recently reported the expression cloning, from rat and opossum cells, of closely related cDNAs encoding receptors for PTH [and PTH-related peptide (PTHRP)]. To determine if these cloned PTH/PTHRP receptors can activate multiple intracellular effectors when present at near-physiological levels in intact target cells, we have stably expressed the rat and opossum PTH/PTHRP receptor cDNAs in LLC- PK1 porcine renal epithelial cells. These cells lack endogenous PTH/PTHRP receptors, but do express abundant calcitonin receptors and many features of a proximal tubular phenotype. Subclones of transfected LLC-PK1 cells exhibited high affinity binding (Kd, 1-5 nM) of [Nle8.18,Tyr34]bovine PTH-(1-34)amide (PTH) and dose-dependent activation by PTH of both cAMP accumulation (EC50, 1 nM) and increased release of cytosolic free calcium from intracellular stores (EC50, > or = 20-50 nM) across a wide range of receptor expression. Expressed rat and opossum receptors exhibited similar properties, except for a 5-fold lower binding affinity of the rat receptor for PTH-(7-34). Stimulation by PTH of both cAMP accumulation and elevated cytosolic free calcium was augmented in cells expressing higher numbers of PTH/PTHRP receptors. Like calcitonin, PTH (1-100 nM) reduced the rate of cell proliferation and augmented the rate of inorganic phosphate transport after 24 and 5 h of preincubation, respectively. The growth effect was mimicked by cAMP analogs, forskolin, phorbol esters, and calcium ionophores. Regulation of phosphate transport, however, was mimicked by phorbols, but not by cAMP analogs or forskolin. We conclude that LLC- PK1 cells provide a useful model in which to study the function of cloned PTH/PTHRP receptors. In these cells, a single species of cloned PTH/PTHRP receptors, stably expressed at near-physiological numbers, activates multiple second messenger responses and regulates subsequent biological responses, including at least one (phosphate transport) that is mediated by mechanisms independent of cAMP.
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T. B. Usdin, C. Gruber, and T. I. Bonner Identification and Functional Expression of a Receptor Selectively Recognizing Parathyroid Hormone, the PTH2 Receptor J. Biol. Chem., June 30, 1995; 270(26): 15455 - 15458. [Abstract] [Full Text] [PDF] |
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A. Iida-Klein, J. Guo, L. Y. Xie, H. Jüppner, J. T. Potts Jr., H. M. Kronenberg, F. R. Bringhurst, A. B. Abou-Samra, and G. V. Segre Truncation of the Carboxyl-terminal Region of the Rat Parathyroid Hormone (PTH)/PTH-related Peptide Receptor Enhances PTH Stimulation of Adenylyl Cyclase but Not Phospholipase C J. Biol. Chem., April 14, 1995; 270(15): 8458 - 8465. [Abstract] [Full Text] [PDF] |
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E Schipani, K Kruse, and H Juppner A constitutively active mutant PTH-PTHrP receptor in Jansen-type metaphyseal chondrodysplasia Science, April 7, 1995; 268(5207): 98 - 100. [Abstract] [PDF] |
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T. J. Gardella, M. D. Luck, A. K. Wilson, H. T. Keutmann, S. R. Nussbaum, J. T. Potts Jr., and H. M. Kronenberg Parathyroid Hormone (PTH)-PTH-related Peptide Hybrid Peptides Reveal Functional Interactions between the 1-14 and 15-34 Domains of the Ligand J. Biol. Chem., March 24, 1995; 270(12): 6584 - 6588. [Abstract] [Full Text] [PDF] |
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C. Zierold, J. A. Mings, and H. F. DeLuca Parathyroid hormone regulates 25-hydroxyvitamin D3-24-hydroxylase mRNA by altering its stability PNAS, November 20, 2001; 98(24): 13572 - 13576. [Abstract] [Full Text] [PDF] |
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C. Marfella-Scivittaro, A. Quinones, and S. A. Orellana cAMP-dependent protein kinase and proliferation differ in normal and polycystic kidney epithelia Am J Physiol Cell Physiol, April 1, 2002; 282(4): C693 - C707. [Abstract] [Full Text] [PDF] |
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