Localization and regulation of a functional GHRH receptor in the rat renal medulla

Endocrinology. 2002 Apr;143(4):1475-84. doi: 10.1210/endo.143.4.8746.

Abstract

To provide information about the kidney GHRH receptor (GHRH-R), we assessed its tissue and cellular localization, defined its pattern of expression in developing and aging rats, and studied the effects of GHRH on the regulation of GHRH-R mRNA levels and receptor internalization. In situ hybridization and ribonuclease protection assay demonstrated that GHRH-R mRNA is restricted to the Henle's loop (HL). GHRH-R mRNA levels were low in the medulla from 3- and 12-d-old male rats, increased significantly in that from 30- to 70-d-old rats, and decreased in that from 12- and 18-month-old animals. Compared with the GHRH-R mRNA profile obtained in the pituitary, these data support the concept of a tissue-specific regulation of GHRH-R. In HL cell cultures from 70-d-old rats, a 4-h incubation with 1-100 nM rat GHRH-(1-29)NH(2) reduced GHRH-R mRNA levels significantly. As anti-GHRH-R- (392-404) immunoreactivity was demonstrated in HL cells, internalization of [N(alpha)-5-carboxyfluoresceinyl-D-Ala(2),Ala(8), Ala(15),Lys(22)]hGHRH-(1-29)NH(2) in a time- and temperature-dependent manner and inhibition of this process by phenyl arsine oxide indicate that desensitization to GHRH involves both GHRH-R internalization and down-regulation of GHRH-R mRNA levels. Localization of a functional GHRH-R in HL and its regulation during development and aging suggest roles associated with cellular proliferation, differentiation, and/or water/electrolyte transport.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Northern
  • Cell Separation
  • Female
  • Gene Expression Regulation / genetics*
  • Immunohistochemistry
  • In Situ Hybridization
  • In Vitro Techniques
  • Kidney Medulla / anatomy & histology
  • Kidney Medulla / growth & development
  • Kidney Medulla / metabolism*
  • Kidney Tubules, Collecting / cytology
  • Kidney Tubules, Collecting / metabolism
  • Loop of Henle / cytology
  • Loop of Henle / metabolism
  • Male
  • Nuclease Protection Assays
  • Pituitary Gland, Anterior / growth & development
  • Pituitary Gland, Anterior / metabolism
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / genetics
  • Rats
  • Rats, Sprague-Dawley
  • Receptors, LHRH / biosynthesis*
  • Receptors, LHRH / genetics

Substances

  • RNA, Messenger
  • Receptors, LHRH