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S-M Luoh, F Di Marco, N Levin, M Armanini, M-H Xie, C Nelson, G L Bennett, M Williams, S A Spencer, A Gurney, and F J de Sauvage

ABSTRACT

Leptin, the product of the ob gene, is a hormone secreted by fat cells which is primarily involved in the regulation of body weight. We have generated a leptin immunoadhesin (leptin-IgG) which was more potent than leptin alone at reducing body weight and food intake when injected into ob/ob mice. This molecule was used to identify high affinity binding sites on human embryonic 293 kidney cells and subsequently to isolate a cDNA encoding the leptin receptor from this cell line by expression cloning. This receptor corresponds to the short form of the recently isolated leptin receptor. Analysis of the expression pattern of the two forms of receptor by Northern blot, in situ hybridization and quantitative PCR showed that the receptor is expressed in most tissues but that the long form is prevalent in the hypothalamus.

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T E Spencer, N H Ing, T L Ott, J S Mayes, W C Becker, G H Watson, M A Mirando, and F W Bazer

ABSTRACT

This study determined the effects of intrauterine injections of recombinant ovine interferon-τ (roIFN-τ; 2 × 107 antiviral units/day) or control proteins (6 mg/day) from day 11 to day 14 post-oestrus (oestrus=day 0) on endometrial expression of receptors for oestrogen, progesterone and oxytocin in cyclic ewes. Plasma concentrations of progesterone were greater on day 15 in ewes receiving roIFN-τ compared with control proteins (P<0·02, treatment × day). Ewes injected with roIFN-τ had lower endometrial levels of oestrogen receptor mRNA (P<0·10) and protein (P<0·01) on day 15 compared with ewes receiving control proteins. In situ hybridization analysis indicated that oestrogen receptor mRNA was more abundant in the luminal and glandular epithelium of control ewes compared with roIFN-τ-treated ewes. Immunoreactive oestrogen receptor was also present in the luminal and glandular epithelium of control, but not roIFN-τ-treated ewes. Endometrial levels of progesterone receptor mRNA and protein were not different (P>0·10) between control and roIFN-τ-treated ewes. In situ hybridization analyses indicated that progesterone receptor mRNA abundance was low in endometrial epithelium and stroma of both control and roIFN-τ-injected ewes. Immunoreactive progesterone receptors were present in the endometrial stroma and epithelium of control ewes, but confined to the stroma of roIFN-τ-treated ewes. Oxytocin receptor density was lower (P<0·10) in the endometrium of ewes injected with roIFN-τ than control proteins; however, oxytocin receptor affinity was not affected (P>0·10) by treatment. Concentrations of 13,14-dihydro-15-keto-prostaglandin F (PGFM) were not increased by exogenous oxytocin administration in control and roIFN-τ-treated ewes on days 10 or 12 post-oestrus. However, on day 14, control ewes responded to oxytocin with increased plasma concentrations of PGFM, whereas ewes receiving roIFN-τ remained unresponsive to oxytocin. These results indicate that the antiluteolytic effects of IFN-τ are to prevent increases in endometrial oestrogen receptor mRNA and protein and oxytocin receptor density which abrogates uterine release of prostaglandin F during maternal recognition of pregnancy. IFN-τ may inhibit the synthesis of oestrogen receptor mRNA by a transcriptional or post-transcriptional regulatory mechanism to suppress oxytocin receptor formation during early pregnancy in ewes.