Ob receptor in rabbit ovary and leptin in vitro regulation of corpora lutea

J Endocrinol. 2004 Nov;183(2):279-88. doi: 10.1677/joe.1.05507.

Abstract

We studied leptin involvement in rabbit corpora lutea (CL) activity, and its post-transcriptional signalling pathway. The expression of leptin receptor (Ob-R) in rabbit ovary at day 9 of pseudopregnancy was evaluated by immunohistochemistry and Western blot analysis. The specificity of the Ob-R receptor antibodies was characterised by immunoprecipitation and competition with blocking peptide. Day 9 CL were incubated in vitro with leptin alone or with inhibitors of PLC (phospholipase C), PLD (phospholipase D), AC (adenylate cyclase), JAK (janus kinase), MAPK (mitogen-activated protein kinase) and both cAMP- and cGMP-specific PDE (phosphodiesterase). Prostaglandin F2alpha(PGF2alpha), PGE2 and progesterone levels were measured in the culture medium, while NOS (nitric oxide synthase) and cAMP- and cGMP- specific PDE activities were measured in CL tissue. Positive staining for Ob-R was found within the cytoplasm of large luteal cells of CL as well as in granulosa cells of follicles and oocytes. Immunoblots detected a band of about 99 kDa size in Ob-R immunoprecipitates from CL homogenates. This band was not detectable after pre-incubation of the primary antibody with the immunising leptin peptide. Leptin increased PGF2alphaand cAMP-specific PDE, decreased basal progesterone and did not affect PGE2 and NOS levels. Leptin used the JAK pathway in increasing PGF2alpha, and MAPK and cAMP-specific PDE in decreasing progesterone. This study supports a permissive luteolytic role for leptin in rabbit CL.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western / methods
  • Corpus Luteum / drug effects*
  • Dinoprost / analysis
  • Female
  • Immunohistochemistry / methods
  • Leptin / pharmacology*
  • Nitric Oxide Synthase / analysis
  • Ovary / chemistry*
  • Phosphoric Diester Hydrolases / analysis
  • Progesterone / analysis
  • Pseudopregnancy
  • Rabbits
  • Receptors, Cell Surface / analysis*
  • Receptors, Leptin
  • Tissue Culture Techniques

Substances

  • Leptin
  • Receptors, Cell Surface
  • Receptors, Leptin
  • Progesterone
  • Dinoprost
  • Nitric Oxide Synthase
  • Phosphoric Diester Hydrolases