Analysis of Steroid‐Induced Genes in the Rat Preoptic Area‐Anterior Hypothalamus Using a Differential‐Display Reverse Transcriptase‐Polymerase Chain Reaction

Abstract

Steroid hormones modulate a variety of physiological functions in the hypothalamus. We attempted to identify steroid-regulated genes in the rat preoptic area-anterior hypothalamus by comparing differentially expressed mRNAs. Adult female rats were ovariectomized and, 1 week later, a silastic capsule containing 17beta-oestradiol (180 microg/ml) was subcutaneously implanted. After 2 days, a single injection of progesterone (1 mg) was administered at 10.00 h and rats were killed at 17.00 h on the same day. Differential-display polymerase chain reaction followed by Northern blot analysis showed that 10 clones were differentially regulated. Using homology search in Genbank, three genes were identified as sodium, potassium-ATPase beta1, protein kinase C-binding Nell-homologue protein and evectin-1. Further characterization of 10 clones showed that the expression patterns were tissue-specific and differentially regulated during puberty. Among these, mRNAs for protein kinase C-binding Nell-homologue protein, evectin-1 and human CGI-118 protein-like gene were induced after vagina opening, and differentially expressed during the oestrous cycle. Taken together, several steroid-regulated genes identified in the present study may play an important role in regulating hypothalamic functions, including puberty and the oestrous cycle.