Analysis of steroid-induced genes in the rat preoptic area-anterior hypothalamus using a differential-display reverse transcriptase-polymerase chain reaction.
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.
Park
S
S
School of Biological Sciences, Seoul National University, Seoul 151-742, Korea.
Seong
J Y
JY
Son
G H
GH
Kang
S S
SS
Lee
S
S
Kim
S R
SR
Kim
K
K
D000818:Animals; D015152:Blotting, Northern; D004343:Drug Implants; D004958:Estradiol / Q000494:pharmacology; D004971:Estrus; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects*; D007032:Hypothalamus, Anterior / Q000737:chemistry / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics; D010052:Ovariectomy; D011301:Preoptic Area / Q000737:chemistry / Q000378:metabolism*; D011374:Progesterone / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017385:Sequence Homology; D012741:Sexual Maturation; D000254:Sodium-Potassium-Exchanging ATPase / Q000235:genetics; D013256:Steroids / Q000494:pharmacology*
D004343:Drug Implants; D008565:Membrane Proteins; C485942:Plekhb1 protein, rat; D012333:RNA, Messenger; D013256:Steroids; D011374:Progesterone; D004958:Estradiol; D011493:Protein Kinase C; D000254:Sodium-Potassium-Exchanging ATPase