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雌激素依赖性猪输卵管中促黄体生成素/人绒毛膜促性腺激素受体的表达及其在输卵管松弛中的作用。

Oestrogen-dependent expression of LH/hCG receptors in pig Fallopian tube and their role in relaxation of the oviduct.

作者信息

Gawronska B, Paukku T, Huhtaniemi I, Wasowicz G, Ziecik A J

机构信息

Institute of Animal Reproduction and Food Research of Polish Academy of Sciences, Olsztyn, Poland.

出版信息

J Reprod Fertil. 1999 Mar;115(2):293-301. doi: 10.1530/jrf.0.1150293.

Abstract

The current studies investigated the concentration and distribution of LH receptors in the oviduct of ovariectomized gilts at various times after administration of oestradiol benzoate (10 micrograms kg-1 body weight) to determine whether LH participates in the regulation of oviductal contractions. Polyclonal antibodies to the LH receptor were used in immunocytochemical and western blot analyses of oviductal tissues. The mechanical activity of the isthmus and ampullar segments of oviduct, collected from 16 cyclic gilts, was recorded for 30 min after LH or hCG treatment. In the oviduct, there was little competition for receptor occupancy between hCG and pig FSH, bovine thyroid-stimulating hormone (TSH), pig growth hormone (GH) and pig prolactin (1.2, 0.1, 0.01 and < 0.001%, respectively) but pig LH could completely inhibit the binding of [125I]hCG. Oestradiol benzoate increased (P < 0.01) the number of LH binding sites in oviduct 24, 48 and 72 h (0.60 +/- 0.08, 1.62 +/- 0.15, 2.48 +/- 0.35 fmol mg-1 protein; n = 4 per treatment, respectively) after injection compared with the control gilts treated with corn oil (0.20 +/- 0.04 fmol mg-1 protein; n = 4). The affinity of oviductal LH/hCG binding sites (Ka) varied from 4.0 to 8.5 x 10(10) l mol-1 and was similar to that of luteal cell binding sites (6.1 x 10(10) l mol-1). Oestradiol benzoate also resulted in more intense LH receptor immunostaining of the tubal mucosal epithelium, smooth muscle cells and blood vessels as compared with controls. Western blotting has revealed that the pig oviduct, similar to the corpus luteum, contains 75, 48 and 45 kDa immunoreactive LH receptor proteins. Treatment with LH in vitro (100 ng ml-1) affected the contractility of oviduct. During the peri-ovulatory stage of the oestrous cycle, the amplitude, frequency and area under curve(s) of the isthmus decreased (P < 0.05), as did the frequency and area under curve (P < 0.05 and P < 0.01, respectively) of the ampulla (n = 4). The frequency and area under curve of the oviductal contractions were also significantly reduced during the early follicular phase of the oestrous cycle (P < 0.05). There was no effect of LH (or hCG) on the frequency and area under curve of the oviductal contractions during luteal stages of the oestrous cycle (n = 8). These data indicate that (1) the pig oviduct possesses immunoreactive and functional LH receptor, (2) oestradiol promotes the synthesis of LH receptor in the epithelium and smooth muscles, and (3) LH causes the relaxation of oviduct, especially during the peri-ovulatory stage of the oestrous cycle. In summary, the results of the present study indicate that LH can control oviductal contractions directly and may be partially responsible for the relaxation of isthmus during fertilization in pigs.

摘要

本研究调查了在给去卵巢后备母猪注射苯甲酸雌二醇(10微克/千克体重)后的不同时间,其输卵管中促黄体生成素(LH)受体的浓度和分布,以确定LH是否参与输卵管收缩的调节。采用针对LH受体的多克隆抗体对输卵管组织进行免疫细胞化学和蛋白质印迹分析。记录了从16头发情周期正常的后备母猪采集的输卵管峡部和壶腹部节段在LH或人绒毛膜促性腺激素(hCG)处理后30分钟的机械活性。在输卵管中,hCG与猪促卵泡素(FSH)、牛促甲状腺激素(TSH)、猪生长激素(GH)和猪催乳素之间对受体占据几乎没有竞争(分别为1.2%、0.1%、0.01%和<0.001%),但猪LH可完全抑制[125I]hCG的结合。与用玉米油处理的对照后备母猪(0.20±0.04飞摩尔/毫克蛋白质;n = 4)相比,注射苯甲酸雌二醇后24、48和72小时,输卵管中LH结合位点的数量增加(P < 0.01)(分别为0.60±0.08、1.62±0.15、2.48±0.35飞摩尔/毫克蛋白质;每个处理n = 4)。输卵管LH/hCG结合位点的亲和力(Ka)在4.0至8.5×10¹⁰升/摩尔之间,与黄体细胞结合位点的亲和力(6.1×10¹⁰升/摩尔)相似。与对照相比,苯甲酸雌二醇还导致输卵管黏膜上皮、平滑肌细胞和血管的LH受体免疫染色更强。蛋白质印迹显示,猪输卵管与黄体一样,含有75、48和45千道尔顿的免疫反应性LH受体蛋白。体外给予LH(100纳克/毫升)会影响输卵管的收缩性。在发情周期的排卵前期,峡部的振幅、频率和曲线下面积降低(P < 0.05),壶腹部的频率和曲线下面积也降低(分别为P < 0.05和P < 0.01)(n = 4)。在发情周期的黄体期,LH(或hCG)对输卵管收缩的频率和曲线下面积没有影响(n = 8)。这些数据表明:(1)猪输卵管具有免疫反应性和功能性LH受体;(2)雌二醇促进上皮和平滑肌中LH受体的合成;(3)LH导致输卵管松弛,尤其是在发情周期的排卵前期。总之,本研究结果表明LH可直接控制输卵管收缩,可能部分负责猪受精期间峡部的松弛。

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