血管紧张素II受体亚型在大鼠睾丸发育中的作用

H. KANEHARA, K. SONG, K. HIRAI, H. UEDA, N. SHIOTA, H. AZUMA, Y. KATSUOKA, H. MIYAZAKI, M. MIYAZAKI
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引用次数: 14

摘要

采用体外放射自显像和Northern blot方法研究了Sprague-Dawley大鼠在不同发育阶段(出生后1、5天、2、3、4、7周)睾丸血管紧张素II (AT2)受体的表达。受体以125I-[Sar1, Ile8]AT2标记,并根据其对AT1(氯沙坦,5 μM)和AT2 (PD123319, 5 μM)拮抗剂的敏感性分为两种亚型。睾丸AT2受体结合总量在1日龄时最高(8.12±0.35 fmol/mg蛋白,平均±secEM, n = 8),此后逐渐降低(5天:6.9±0.41,2周:2.85±0.10,3周:1.64±0.19,4周:0.76±0.09,6周:0.77±0.09 fmol/mg蛋白,n = 8 - 11)。1日龄大鼠睾丸中AT2受体结合明显丰富(6.98±0.34 fmol/mg蛋白),而AT1受体结合明显较少(1.46±0.19 fmol/mg蛋白)。每种亚型的相对数量在前3周内没有变化,但4周龄大鼠睾丸几乎完全含有AT1受体(0.63±0.05 fmol/mg蛋白)。Northern blot分析显示,AT1和AT2型mRNA的表达随年龄的增长而下降。显微乳化液放射自显影以明确结合的定位。10日龄时,间质区同时存在AT1和AT2受体,而精小管中主要含有AT2受体。7周龄时,精小管未见明显结合,间质区仅含有AT1受体。这些结果表明AT2受体在快速生长的睾丸中表达,并提示AT2受体亚型水平的变化可能与睾丸的发育和/或生长有关。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Involvement of angiotensin II receptor subtypes during testicular development in rats

Involvement of angiotensin II receptor subtypes during testicular development in rats

Expression of testicular angiotensin II (AT2) receptors in Sprague–Dawley rats at various stages of development (1 and 5 days, 2, 3, 4 and 7 weeks postnatal) were studied by in vitro autoradiography and Northern blot analysis. The receptors were labelled with 125I-[Sar1, Ile8]AT2 and differentiated into two subtypes according to their susceptibility to AT1 (losartan, 5 μM) or AT2 (PD123319, 5 μM) antagonists. Total AT2 receptor binding in the testis was highest at 1 day of age (8.12 ± 0.35 fmol/mg protein, mean ± secEM, n = 8) and decreased gradually thereafter (5 days: 6.9 ± 0.41, 2 weeks: 2.85 ± 0.10, 3 weeks: 1.64 ± 0.19, 4 weeks: 0.76 ± 0.09, 6 weeks: 0.77 ± 0.09 fmol/mg protein, n = 8–11). AT2 receptor binding was strikingly abundant in 1-day-old rat testis (6.98 ± 0.34 fmol/mg protein), while considerably less AT1 receptor binding (1.46 ± 0.19 fmol/mg protein) was observed. The relative amounts of each subtype did not change for the first 3 weeks but the 4-week-old rat testis contained almost exclusively AT1 receptors (0.63 ± 0.05 fmol/mg protein). Northern blot analysis showed that mRNA expression of both AT1 and AT2 types decreased with age. Microscopic emulsion autoradiography was undertaken to clarify the localization of binding. At 10 days of age, both AT1 and AT2 receptors were present in the interstitial area, whereas seminiferous tubules contained mainly AT2 receptors. At 7 weeks of age, no significant binding was observed in the seminiferous tubule and the interstitial area contained AT1 receptors exclusively. These results demonstrate expression of AT2 receptors in the rapidly growing testis and suggest that change in the levels of AT2 receptor subtypes may be relevant to development and/or growth of the testis.

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