Ottobre J S, Eyster K M, Stouffer R L
Biol Reprod. 1984 Mar;30(2):271-7. doi: 10.1095/biolreprod30.2.271.
In polyovular species, it is unclear whether the characteristics of each individual corpus luteum (CL), such as mass, progesterone concentration and receptors for luteinizing hormone (LH), are representative of those of its cohorts during the ovarian cycle. The current study was performed 1) to characterize the conditions for estimation of binding parameters for LH receptors in porcine CL, and 2) to compare LH binding sites, luteal progesterone concentrations and luteal masses among CL of ovaries within individual pigs. Gonadotropin binding sites in porcine CL were characterized via specific binding of 125I-human (h) LH to 20,000 X g particulate fractions of luteal tissue. Specific binding was directly proportional to tissue content and was detectable at the lowest content tested (0.5 mg tissue equivalents/tube). Specific uptake of 0.25 ng LH by 5.0 mg tissue equivalents was time- and temperature-dependent; steady-state binding was achieved within 20 h at 37 and 25 degrees C. Binding of LH after 20 h incubation at 37 degrees C (4718 +/- 192 cpm, means +/- SEM) and 25 degrees C (4112 +/- 340 cpm) was greater than that at 4 degrees C (1930 +/- 5 cpm, P less than 0.01). Luteal particulates from individual CL of ovaries collected from four mature nonpregnant pigs (13-23 CL/pig) were incubated with eight concentrations of 125I-hLH. Steady-state binding depended upon hormone concentration until reaching saturation at 2.5 ng 125I-hLH/tube. Scatchard analyses yielded linear plots. Binding capacities for LH ranged among pigs from 0.71 +/- 0.03 to 3.69 +/- 0.13 fmol/mg CL equivalents and receptor affinities (Kd) ranged from 0.92 +/- 0.05 to 4.89 +/- 0.41 X 10(-11) M.(ABSTRACT TRUNCATED AT 250 WORDS)
在多卵性物种中,尚不清楚每个黄体(CL)的特征,如质量、孕酮浓度和促黄体生成素(LH)受体,在卵巢周期中是否代表其同组黄体的特征。本研究的目的是:1)确定猪黄体中LH受体结合参数的估计条件;2)比较个体猪卵巢内黄体之间的LH结合位点、黄体孕酮浓度和黄体质量。通过将125I-人(h)LH与黄体组织的20,000×g颗粒部分进行特异性结合,来表征猪黄体中的促性腺激素结合位点。特异性结合与组织含量成正比,在测试的最低含量(0.5毫克组织当量/管)时即可检测到。5.0毫克组织当量对0.25纳克LH的特异性摄取具有时间和温度依赖性;在37℃和25℃下20小时内达到稳态结合。在37℃(4718±192 cpm,平均值±标准误)和25℃(4112±340 cpm)孵育20小时后LH的结合量大于4℃(1930±5 cpm,P<0.01)。从四只成熟未孕猪(每头猪13 - 23个黄体)收集的卵巢中,将单个黄体的颗粒与八种浓度的125I-hLH一起孵育。稳态结合取决于激素浓度,直到在2.5纳克125I-hLH/管时达到饱和。Scatchard分析产生线性图。猪之间LH的结合能力范围为0.71±0.03至3.69±0.13飞摩尔/毫克黄体当量,受体亲和力(Kd)范围为0.92±0.05至4.89±0.41×10-11 M。(摘要截断于250字)