IMMUNOHISTOCHEMICAL LOCALIZATION OF ANDROGEN RECEPTORS IN THE RAT TESTIS - EVIDENCE FOR STAGE-DEPENDENT EXPRESSION AND REGULATION BY ANDROGENS
IMMUNOHISTOCHEMICAL LOCALIZATION OF ANDROGEN RECEPTORS IN THE RAT TESTIS - EVIDENCE FOR STAGE-DEPENDENT EXPRESSION AND REGULATION BY ANDROGENS
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DOI：
10.1210/en.135.3.1227
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发表时间：
1994-09-01
期刊：
影响因子：
4.8
通讯作者：
SAUNDERS, PTK
中科院分区：
文献类型：
--
作者：
BREMNER, WJ;MILLAR, MR;SAUNDERS, PTK
雄激素对维持大鼠正常的精子发生是必不可少的。我们评估了大鼠睾丸中雄激素受体(AR)的位置、发育模式和激素控制。成年雄性大鼠在1)不处理；2)乙烷二甲基磺酸盐(EDS)，它根除间质细胞和内源性睾酮(T)；3)EDS加T在给药时开始替换；或4)甲氧基乙酸，导致特定生殖细胞类型的丧失后进行研究。取正常未成熟大鼠(5、14、16、21、28、31、35、38和45d龄)的睾丸。微波抗原修复后，用针对AR N-末端的多肽的兔多克隆抗体(Novocasta)和生物素标记的猪抗兔免疫球蛋白G、生物素-生物素复合体/碱性磷酸酶、硝基蓝四氮唑盐(NBT)/5-溴-4-氯-3-吲哚基磷酸(BCIP)底物进行免疫组织化学检测。在成人，从生精周期的第二阶段到第七阶段，支持细胞(SC)的核免疫染色逐渐增强，然后在第八阶段急剧下降，到第九阶段到第十三阶段几乎检测不到。肾小管周围肌样细胞、小动脉和间质细胞也有明显的AR免疫染色；这些细胞的染色不随相邻小管周期的不同而变化。EDS导致所有细胞类型的AR免疫染色严重丧失。在EDS处理的动物中，T的替换导致AR免疫染色的模式与对照组相似，尽管染色强度降低。甲氧基乙酸给药不影响AR染色模式。幼年大鼠肾小管周围肌样细胞免疫染色从第5天开始明显，第5天可见SC染色，染色强度随年龄增加而增强，21~35天呈阶段依赖性。1)免疫组织化学显示，AR在SC中的表达主要发生在生精周期的II-VII期，在VII期表达最高。2)AR免疫染色在肾小管周围肌样细胞、小动脉和间质细胞中也有明显表达(但在生殖细胞中不明显)，但与邻近小管的分期无关。3)内源性T和/或其代谢产物控制AR在睾丸的表达。4)AR免疫染色在5日龄时可检测到，并在第21-35日龄出现阶段特异性。
Androgens are essential for the maintenance of normal spermatogenesis in the rat. We assessed the sites, developmental pattern, and hormonal control of androgen receptors (AR) in the rat testis. Adult male rats were studied after 1) no treatment; 2) ethane dimethane sulfonate (EDS), which eradicates Leydig cells and endogenous testosterone (T); 3) EDS plus T replacement beginning at the time of EDS administration; or 4) methoxyacetic acid, which leads to the loss of specific germ cell types. Testes were also obtained from normal immature rats (aged 5, 14, 16, 21, 28, 31, 35, 38, and 45 days). After microwave antigen retrieval, immunohistochemistry was performed using a rabbit polyclonal antibody (Novocastra) raised against a peptide unique to the N-terminal region of the AR and detection with biotinylated swine antirabbit immunoglobulin G, avidin-biotin complex/ alkaline phosphatase, and nitroblue tetrazolium salt (NBT)/5 bromo-4-chloro-3-indolylphosphate (BCIP) substrate. In adults, nuclear immunostaining of Sertoli cells (SC) increased progressively in intensity from stages II through VII of the spermatogenic cycle, and then declined precipitously during stage VIII to become barely detectable in stages IX-XIII. Prominent AR immunostaining was also evident in peritubular myoid cells, arterioles, and interstitial cells; staining in these cells did not vary with the stage of the cycle of the adjacent tubules. EDS caused a severe loss of AR immunostaining in all cell types. Replacement of T in EDS-treated animals resulted in a pattern of AR immunostaining comparable to that in controls, although staining intensity was reduced. Methoxyacetic acid administration did not affect the pattern of AR staining. In immature rats, peritubular myoid cell immunostaining was prominent from day 5; SC staining was detectable on day 5, increased in intensity with age, and became stage dependent between days 21-35.The following conclusions were reached. 1) Immunohistochemically detectable AR expression in SC occurs predominantly in stages II-VII of the spermatogenic cycle, with highest levels at stage VII. 2) AR immunostaining is also prominent in peritubular myoid cells, arterioles, and Leydig cells (but not in germ cells), but is unrelated to the stage of adjacent tubules. 3) Endogenous T and/or its metabolites control the expression of AR in the testis. 4) AR immunostaining is detectable by day 5 of age and becomes stage specific in SC between days 21-35.