An improved method for preparation of follicle stimulating and luteinizing hormones from horse pituitary glands

Abstract

Finely ground fresh frozen horse pituitaries were extracted with 33% ethanol, and the extract was adjusted to pH 6 with 0.1 N HCl. The proteins in the supernatant containing the gonadotropic activity were precipitated with 2 vol. of acetone. This precipitate could be separated into FSH (follicle stimulating hormone) and LH (luteinizing hormone) fractions by zone electrophoresis. The FSH fraction was further separated into FSH and LH by starch-gel electrophoresis and both FSH and LH eluted by a second electrophoresis in an elution cell. Soluble starch from the FSH and LH preparations was removed by ion-exchange chromatography on Dowex-2 X8. In normal rats FSH equivalent to 2 and 5 g fresh tissue produced 80 and 119 mg ovaries, respectively; whereas in hypophysectomized rats these doses produced 57 and 71 mg ovaries. The ovaries of the hypophysectomized rats contained follicles and uteri were distended with fluid indicating estrogen production. FSH equivalent to 2 and 5 g fresh tissue stimulated the ventral prostates of normal and hypophysectomized rats little, if any, but FSH equivalent to 10 g fresh tissue increased the weights of the ventral prostates of the hypophysectomized rats from 11 to 37 mg. Doses of LH equivalent to 2 and 5 g fresh tissue in normal and 2, 5 and 10 g fresh tissue in hypophysectomized rats significantly stimulated the ventral prostates but did not stimulate the ovaries significantly. LH also stimulated testes and seminal vesicles. Two distinct LH entities were identified.