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a Prince Henry's Institute of Medical Research, Clayton, Victoria 3168, Australia
Establishment of receptive endometrium is essential for implantation. Our aim was to identify and characterize genes uniquely regulated at the sites of implantation in mouse uterus by RNA differential display polymerase chain reaction (DDPCR). One of the gene fragments identified was 86% homologous to rat calcium-binding protein D9k (calbindin-D9k); the mouse counterpart had not then been cloned, but subsequently an mRNA sequence of mouse calbindin-D9k became available in GenBank (accession number: AF028071). This sequence is 99% homologous to the DDPCR-derived gene tag but has a shorter 3' end. Reverse transcription-polymerase chain reaction (RT-PCR) was performed using the sequence of 3' end of the DDPCR product and the 5' end of AF028071, and a full cDNA was obtained. This gene was primarily up-regulated by progesterone, but not by estrogen. It was further increased by the combination of the two steroids. Expression of calbindin-D9k was overall increased in the uterus during early pregnancy, but the level was significantly lower in implantation compared to interimplantation sites on Days 4.5 and 5.5 of pregnancy, becoming barely detectable in both sites after Day 6.5. In situ hybridization localized this mRNA predominantly in the luminal epithelium of the pregnant uterus. The complex regulation of calbindin-D9k in mouse uterus suggests an important role for this protein during pregnancy.
1 Research supported by the Rockefeller Foundation Contraceptive 21 Initiative Project, the Wellcome Trust (grant 52666), and the NH&MRC of Australia (grant 971297).
2 Correspondence: Gui-Ying Nie, Prince Henry's Institute of Medical Research, PO Box 5152, Block E, Level 4, Monash Medical Centre, 246 Clayton Rd, Clayton, Vic 3168, Australia. FAX: 61 3 9594 6125; guiying.nie{at}med.monash.edu.au
3 Current address: Shanghai Institute of Planned Parenthood Research, 2140 Xie Tu Road, Shanghai 200032, China.
4 Current address: Department of Obstetrics and Gynecology, Mie University School of Medicine, 2-174 Edobashi Tsu, Mie, 514 Japan.
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