Kennedy T G, Keys J L, King G J
Biol Reprod. 1986 Oct;35(3):624-32. doi: 10.1095/biolreprod35.3.624.
Because prostaglandins (PGs) are thought to mediate, in part, endometrial responses at implantation in a number of species and because pig conceptuses are capable of producing PGs, we have characterized the binding of [3H]PGE2 to endometrium obtained from pigs during the estrous cycle and early pregnancy. Binding of [3H]PGE2 to a membrane preparation obtained from endometrium was temperature- and pH-dependent, and reversible. Pretreatment of the membrane preparation with proteolytic enzymes, phospholipase A2, or heat reduced subsequent specific binding of [3H]PGE2. PGE1 and PGE2 competed equally for binding, whereas relative cross-reactivity for other prostanoids and compounds tested was less than 5%. Scatchard analysis of specific binding suggested that two classes of binding sites were present, a high-affinity binding site with an apparent dissociation constant (Kd) of approximately equal to 6 pM, and a lower-affinity site with an apparent Kd of approximately equal to 3 nM. For the high-affinity binding site, estimates of apparent Kd did not differ between days of the estrous cycle, but maximum binding capacity did change, being lowest on Day 10 and highest on Days 0-5 of the cycle. For the lower-affinity binding site, estimates of the apparent Kd differed, being lowest on Day 10 and highest on Day 15; numbers of binding sites were lowest on Day 5 and highest on Day 15. Estimates of these variables on Day 15 of pregnancy did not differ significantly from Day 15 of the cycle. These data are consistent with the presence of PGE receptors within the endometrium of the pig.
由于前列腺素(PGs)被认为在许多物种的着床过程中部分介导子宫内膜反应,并且由于猪的胚胎能够产生PGs,我们已经对[3H]PGE2与发情周期和妊娠早期猪子宫内膜的结合特性进行了表征。[3H]PGE2与从子宫内膜获得的膜制剂的结合是温度和pH依赖性的,并且是可逆的。用蛋白水解酶、磷脂酶A2或加热预处理膜制剂会降低随后[3H]PGE2的特异性结合。PGE1和PGE2对结合的竞争程度相同,而其他测试的前列腺素和化合物的相对交叉反应性小于5%。特异性结合的Scatchard分析表明存在两类结合位点,一类高亲和力结合位点的表观解离常数(Kd)约等于6 pM,另一类低亲和力位点的表观Kd约等于3 nM。对于高亲和力结合位点,发情周期各天的表观Kd估计值没有差异,但最大结合容量确实发生了变化,在周期的第10天最低,在第0 - 5天最高。对于低亲和力结合位点,表观Kd的估计值有所不同,在第10天最低,在第15天最高;结合位点数在第5天最低,在第15天最高。妊娠第15天这些变量的估计值与周期第15天没有显著差异。这些数据与猪子宫内膜中存在PGE受体一致。