Identification of relaxin in the placenta of the ewe.

Abstract

Sheep placentomes were collected at the abattoir and the stage of gestation was estimated from the crown-rump length and appearance of the fetus. Samples were extracted and either freeze dried (crude extracts) or fractionated on Sephadex G-50 and CM-cellulose. Relaxin immunoreactivity (RXN-IR) was detected in all samples by a pig relaxin RIA and diluted in parallel with the standard curve. Two patterns of RXN-IR were seen after Sephadex G50 purification: (a) a single main peak of RXN-IR eluting at a position similar to pig relaxin; or (b) a 3-peak pattern with additional higher (void volume) and lower (approximately 1000) molecular weight peaks. These peaks were all found with 4 different and specific antisera. The 6000 molecular weight peak eluted at a similar position to pig relaxin on CM cellulose and inhibited electrically stimulated rat uterine contractions in vitro. The amount of relaxin measured in crude extracts of placentomes from different ewes was very variable. Most samples were within the range 0.05-11.2 ng/g wet weight of tissue (3.0 +/- 0.45 (s.e.m.), n = 44) but a few contained much higher concentrations (25.5-61.4 ng/g, n = 3). There was no obvious variation in concentration with stage of pregnancy (20 days to term). Samples of intercotyledonary endometrium, allantochorion and whole ovaries from pregnant ewes were also extracted. All contained low concentrations of RXN-IR (0.6 +/- 0.13 ng/g, n = 4; 0.6 +/- 0.29 ng/g, n = 3; 1.0 +/- 0.66 ng/g, n = 7, respectively). We conclude that relaxin-like peptides are present in the pregnant ewe and that, as the placentomes are the largest component by weight, they represent the major source.