Porcine calbindin-D9k gene: expression in endometrium, myometrium, and placenta in the absence of a functional estrogen response element in intron A.
The expression of Calbindin-D9k (CaBP-9k) in the pig uterus and placenta was measured by Northern blot analysis and reverse transcription polymerase chain reaction (PCR), respectively. Progesterone (P4) administration to ovariectomized pigs decreased CaBP-9k mRNA levels. Expression of endometrial CaBP-9k mRNA was high on pregnancy Days 10-12 and below the detection limit on Days 15 and 18. On Day 60, expression could be detected at low levels. In myometrium and placenta, CaBP-9k mRNA expression was not detectable by Northern analysis using total RNA. Reverse-transcribed RNA from both tissues demonstrated the presence of CaBP-9k transcripts by means of PCR. The partial CaBP-9k gene was amplified by PCR and cloned to determine the sequence of intron A. In contrast to the rat CaBP-9k gene, the pig gene does not contain a functional estrogen response element (ERE) within this region. A similar ERE-like sequence located at the identical location was examined by gel retardation analysis and failed to bind the estradiol receptor. A similar disruption of this ERE-like sequence has been described in the human CaBP-9k gene, which is not expressed at any level in placenta, myometrium, or endometrium. It is concluded that the pig CaBP-9k gene is regulated in these reproductive tissues in a manner distinct from that in rat and human tissues. The regulation is probably due to a regulatory region outside of intron A, which in the rat gene contains the key cis element for uterine expression of the CaBP-9k gene.
Krisinger
J
J
Department of Obstetrics and Gynecology, University of British Columbia Vancouver, Canada.
Jeung
E B
EB
Simmen
R C
RC
Leung
P C
PC
D000818:Animals; D001483:Base Sequence; D064026:Calbindins; D004247:DNA / Q000235:genetics; D004717:Endometrium / Q000378:metabolism*; D004967:Estrogens / Q000494:pharmacology; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007438:Introns; D008969:Molecular Sequence Data; D009215:Myometrium / Q000378:metabolism*; D010052:Ovariectomy; D010920:Placenta / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011374:Progesterone / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D064030:S100 Calcium Binding Protein G / Q000235:genetics*; D013552:Swine; D013997:Time Factors
D064026:Calbindins; D004967:Estrogens; D012333:RNA, Messenger; D064030:S100 Calcium Binding Protein G; C576506:S100G protein, human; C576508:S100g protein, rat; D011374:Progesterone; D004247:DNA