Vornberger W, Prins G, Musto N A, Suarez-Quian C A
Department of Cell Biology, Georgetown University Medical Center, Washington, D.C. 20007.
Endocrinology. 1994 May;134(5):2307-16. doi: 10.1210/endo.134.5.8156934.
The distribution of the androgen receptor (AR) in the adult rat testis was determined by biotin-streptavidin immunoperoxidase, employing tissue embedded in polyester wax which preserves antigenicity without compromising tissue preservation. The antibody probe used, which has been characterized previously, was an affinity purified, rabbit polyclonal antibody raised to the amino terminus peptide of the rat AR. Within the interstitial compartment, AR immunostaining was detected in some Leydig cells and all smooth muscle cells forming the walls of blood vessels, but endothelial cells of blood vessels were negative. Furthermore, in those Leydig cells that were clearly identified as exhibiting AR immunostaining, the intensity of the reaction varied. In the seminiferous tubules AR immunostaining was observed in all peritubular myoid cell nuclei, but not in the distal layer of lymphatic endothelial cells. In Sertoli cells, nuclear AR immunostaining was stage specific. Moderate AR immunostaining first became evident at late stage IV or early stage V of the cycle, reached a robust peak at stages VII-VIII, and then disappeared completely. Specific AR immunostaining was also discerned in the nuclei of stage XI elongated spermatids, the spermatids in which nuclear elongation is apparent but chromatin condensation has not yet begun. Next, with onset of chromatin condensation, nuclear AR immunostaining in elongated spermatids was not discerned concomitant with its detection in the cytoplasm of the germ cells. These results are interpreted in the following manner: 1) The presence of AR in Leydig cells is consistent with the hypothesis that androgens modify Leydig cell activity in an autocrine fashion. Further, that not all Leydig cells exhibited AR immunostaining at steady state suggests a differential, functional activity of these cells within the population. 2) The intense AR immunostaining of smooth muscle cells present in the interstitium indicates that these cells are targets for androgens. 3) AR immunoreactivity in both Sertoli and peritubular myoid cells suggests their involvement in the androgenic control of spermatogenesis. The stage specific AR immunoreactivity in Sertoli cells, however, may be more indicative of a specific androgen response during these stages, whereas peritubular cells may participate in the tonal maintenance of spermatogenesis. 4) The specific presence of AR in step 11 elongated spermatids may suggest that androgens can act directly on germ cells to regulate spermatogenesis.
采用保存在聚酯蜡中的组织,运用生物素 - 链霉亲和素免疫过氧化物酶法,测定成年大鼠睾丸中雄激素受体(AR)的分布,该方法既能保留抗原性又不影响组织保存。所用抗体探针先前已进行过特性鉴定,是一种针对大鼠AR氨基末端肽产生的亲和纯化兔多克隆抗体。在间质区,在一些睾丸间质细胞和构成血管壁的所有平滑肌细胞中检测到AR免疫染色,但血管内皮细胞呈阴性。此外,在那些明确显示有AR免疫染色的睾丸间质细胞中,反应强度各不相同。在生精小管中,在所有管周肌样细胞核中观察到AR免疫染色,但在远端淋巴管内皮细胞层中未观察到。在支持细胞中,核AR免疫染色具有阶段特异性。中等强度的AR免疫染色在周期的IV期末或V期初首次明显出现,在VII - VIII期达到高峰,然后完全消失。在XI期伸长精子细胞核中也可检测到特异性AR免疫染色,此时精子细胞核伸长明显但染色质浓缩尚未开始。接下来,随着染色质浓缩开始,伸长精子细胞核中的AR免疫染色消失,同时在生殖细胞的细胞质中可检测到AR。这些结果可作如下解释:1)睾丸间质细胞中存在AR与雄激素以自分泌方式调节睾丸间质细胞活性的假说一致。此外,在稳态时并非所有睾丸间质细胞都显示AR免疫染色,这表明这些细胞群体内存在功能差异。2)间质中平滑肌细胞强烈的AR免疫染色表明这些细胞是雄激素的作用靶点。3)支持细胞和管周肌样细胞中的AR免疫反应性表明它们参与雄激素对精子发生的调控。然而,支持细胞中阶段特异性的AR免疫反应性可能更表明这些阶段存在特定的雄激素反应,而管周细胞可能参与精子发生的张力维持。4)在第11步伸长精子细胞中特异性存在AR可能表明雄激素可直接作用于生殖细胞以调节精子发生。