A cloned porcine renal calcitonin receptor couples to adenylyl cyclase and phospholipase C

Am J Physiol. 1992 Jun;262(6 Pt 2):F1110-5. doi: 10.1152/ajprenal.1992.262.6.F1110.

Abstract

The signal transduction pathways of the recently cloned porcine kidney calcitonin (CT) receptor were evaluated. This receptor, when stably transfected into MC-3T3 cells, avidly bound salmon CT (SCT) [dissociation constant (Kd) = 4 nM]. Incubation with SCT resulted in a dose-dependent accumulation of adenosine 3',5'-cyclic monophosphate (cAMP) [50% effective concentration (EC50) = 0.02 nM] in transfected cells (referred to as PC-1 cells). Binding kinetics and cAMP dose response relationships were similar to those of the native receptor in LLC-PK1 cells. PC-1 cells also responded to calcitonin gene-related peptide (CGRP), but the EC50 value for cAMP accumulation was more than three orders of magnitude higher than for SCT. Exposure of PC-1 cells to SCT (5 nM to 1 microM) produced a dose-dependent rise in cytosolic free Ca2+ concentration ([Ca2+]i), whereas CGRP did not. The initial rise in [Ca2+]i was not dependent on extracellular Ca2+, suggesting that SCT induced release of Ca2+ from intracellular stores. SCT also increased inositol trisphosphate production in PC-1 cells. In conclusion, the cloned, transfected porcine CT receptor functionally couples to and activates both adenylyl cyclase and phospholipase C. This dual coupling is also a characteristic of the parathyroid hormone receptor, which has significant homology in amino acid sequence with the CT receptor.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenylyl Cyclases / metabolism*
  • Animals
  • Calcitonin / analysis
  • Calcitonin / pharmacology
  • Calcium / pharmacology
  • Cell Line
  • Cloning, Molecular
  • Culture Media
  • Kidney / cytology
  • Kidney / metabolism*
  • Osmolar Concentration
  • Receptors, Calcitonin
  • Receptors, Cell Surface / genetics
  • Receptors, Cell Surface / metabolism*
  • Salmon / metabolism
  • Signal Transduction
  • Swine
  • Type C Phospholipases / metabolism*

Substances

  • Culture Media
  • Receptors, Calcitonin
  • Receptors, Cell Surface
  • Calcitonin
  • Type C Phospholipases
  • Adenylyl Cyclases
  • Calcium