Dias J A, Reeves J J
J Reprod Fertil. 1982 Sep;66(1):39-45. doi: 10.1530/jrf.0.0660039.
Bulls (N = 42) ranging in age from 1 day to 5.5 years were used to determine whether a change in the concentration of FSH receptors in the bovine testis occurred as bulls matured. 125I-labelled human FSH was used as the ligand to evaluate binding to bovine testicular membranes. Membrane fractions were collected by centrifugation of testicular homogenates at 120 g and recentrifugation of the 120 g supernatant at 1250 g. Relative binding activity of membrane sedimented at 1250 g was determined after incubation of membranes with 125I-labelled FSH for 16--18h at 25 degrees C, followed by centrifugation (1250 g) to separate bound from free hormone. Specifically bound FSH when expressed as fmol/mg protein was negatively correlated with age (r = -0.73). The association constant (Ka) determined by Scatchard analysis was the same for bulls at all ages with a mean (+/- s.e.m.) Ka = 1.5 +/- 0.3 x 10(9) M-1. Concentration of FSH receptors on a per mg protein basis declined rapidly from birth to 2.5 years of age and remained low up to 5.5 years of age. On a whole testis basis the total number of receptors increased as the bulls matured. After 2.5 years of age total testicular binding did not change.
选用年龄从1日龄到5.5岁的公牛(N = 42头)来确定随着公牛成熟,其睾丸中促卵泡素(FSH)受体浓度是否会发生变化。使用125I标记的人促卵泡素作为配体,以评估其与牛睾丸膜的结合情况。通过将睾丸匀浆在120g下离心,然后将120g上清液在1250g下再次离心来收集膜部分。在25℃下将膜与125I标记的促卵泡素孵育16 - 18小时,随后离心(1250g)以分离结合态和游离态激素,之后测定在1250g下沉淀的膜的相对结合活性。以fmol/mg蛋白质表示的特异性结合促卵泡素与年龄呈负相关(r = -0.73)。通过Scatchard分析确定的结合常数(Ka)在所有年龄段的公牛中相同,平均(±标准误)Ka = 1.5 ± 0.3×10(9) M-1。以每毫克蛋白质计算,促卵泡素受体浓度从出生到2.5岁迅速下降,直至5.5岁一直保持在低水平。以整个睾丸计算,随着公牛成熟,受体总数增加。2.5岁后,睾丸总结合量没有变化。