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Development and Application of a Radioimmunoassay for Bovine Follicle-Stimulating Hormone^1,2,

US Department of Agriculture, Beltsville, MD 20705

Abstract

A sensitive, specific and reliable homologous bovine follicle-stimulating hormone (FSH) radioimmunoassay was developed. An antibovine FSH /3-subunit antiserum (USDA-5-0122) and a highly purified, radioiodinated bovine FSH (USDA-FSH-BP1) were used. Cross-reactivities with purified preparations of bovine luteinizing hormone (LH), bovine growth hormone (GH), and bovine prolactin were <.09%. For bovine thyroid-stimulating hormone (TSH), cross-reactivity was .5%. Sensitivity of the assay was about 3 ng relative to the reference standard NIH-FSH-B1. Bovine plasma and bovine FSH inhibited binding in a parallel fashion. However, inhibition curves obtained with human FSH, porcine FSH and several preparations of ovine FSH were not parallel, indicating species differences in FSH. The concentration of FSH in plasma of cows was relatively uniform throughout the estrous cycle, during pregnancy and the first 40 d postpartum. Mean values ranged from 50 ± 5 ng/ml around the time of estrus to 21 ± 5 ng/ml late in pregnancy. Mean FSH values (n = 6 to 16) for 1-mo-old heifers, ovariectomized heifers, bulls and steers were 68 ± 12, 227 ± 19, 31 ± 5 and 136 ± 12 ng/ml, respectively. The concentration of FSH and LH increased in plasma at a similar rate after ovariectomy; about sixfold in 22 d. After administration of gonadotropin-releasing hormone or an analogue of gonadotropin-releasing hormone, FSH increased only marginally, but LH increased about 20-fold.

¹ The authors thank Drs. L. Reichert, Jr., H. Papkoff, J. Pierce, A. Parlow and the National Pituitary Agency for providing purified pituitary hormones and Dr. G. Kiracofe for supplying plasma samples from ovariectomized heifers and sexually mature bulls.

² Mention of a trade name, proprietary product, or specific equipment does not constitute a guarantee or warranty by the USDA and does not imply its approval to the exclusion of other products that may be suitable.

³ Reprod. Lab., Anim. Sci. Institute, Beltsville Agr. Res. Center, ARS.