Koeva Yvetta A, Bakalska Mariana V, Atanassova Ninan N, Davidoff Michail S
Department of Anatomy, Histology and Embryology, Medical University, Plovdiv, Bulgaria.
Folia Med (Plovdiv). 2008 Jul-Sep;50(3):53-7.
The regulatory effect of thyroid hormones on the proliferation and maturation of the Leydig cells (LC) in testis is still poorly understood. To date, it remains obscure whether the thyroid hormones have direct effect on the LC, as far as in rat testis the thyroid hormones receptors are localized predominantly in the Sertoli cells. A single intraperitoneal dose of cytotoxin ethane-1,2-dimethanesulphonate (EDS) injected into mature rats caused a rapid, selective elimination of the adult LC associated with temporary impairment of fertility. Regeneration of the LC population by EDS model is a result of the differentiation of LC progenitors as well as of the proliferation of the newly formed LC whereas the process is similar with the development of adult LC in the prepubertal testis.
The present study aimed to establish the immunohistochemical expression of high affinity triiodothyronine nuclear receptors c-erbAalpha and c-erbAbeta in the regenerating LC after treatment with EDS of mature rats.
Mature male Wistar rats were divided into two groups: (1) a group of rats receiving a single intraperitoneal (i.p.) injection of EDS (75 mg/kg body weight) and (2) a group of control animals. The animals were killed 24 hours, 7, 14, 21 and 35 days after treatment. Testicular fragments were prepared for routine histological and immunohistochemical examinations.
The immunohistochemical analysis revealed similar changes in the immunoreactivity for both c-erbAalpha and c-erbAbeta after EDS administration. On day 1 after EDS treatment, the intensity of the immune reactions for c-erbAalpha and c-erbAbeta in the LCs decreased simultaneously with their number. Seven days after EDS administration there was neither LCs nor c-erbAalpha nor c-erbAbeta-immunoreactivity. The first positive stained LCs were found 14 days after EDS when LCs progenitors were detected. The most prominent c-erbAalpha- and c-erbAbeta-immunostaining in the regenerating LCs was evident 21 days after EDS; this coincided with the increased number of LCs progenitors and their transformation into adult LCs population. Thirty-five days after EDS c-erbAalpha and c-erbAbeta-positive LCs were abundant their number and localization in the testicular interstitium being very similar to that in the control rats.
The observed change in the intensity of the immune reactions for c-erbAalpha and c-erbAbeta in LC repopulation after EDS treatment corresponds to the process of differentiation of progenitors into mature LC. The results obtained support the idea about the regulatory role of thyroid hormones in the differentiation of LC in prepubertal rat testis.
甲状腺激素对睾丸中莱迪希细胞(LC)增殖和成熟的调节作用仍了解甚少。迄今为止,甲状腺激素是否对LC有直接作用仍不清楚,因为在大鼠睾丸中甲状腺激素受体主要定位于支持细胞。向成熟大鼠腹腔内单次注射细胞毒素乙烷-1,2-二甲磺酸盐(EDS)会导致成年LC迅速、选择性地消除,并伴有生育能力的暂时受损。EDS模型中LC群体的再生是LC祖细胞分化以及新形成的LC增殖的结果,而这个过程与青春期前睾丸中成年LC的发育相似。
本研究旨在确定成熟大鼠经EDS处理后再生LC中高亲和力三碘甲状腺原氨酸核受体c-erbAα和c-erbAβ的免疫组化表达。
将成熟雄性Wistar大鼠分为两组:(1)一组大鼠接受腹腔内(i.p.)单次注射EDS(75mg/kg体重);(2)一组为对照动物。在处理后24小时、7天、14天、21天和35天处死动物。制备睾丸组织切片用于常规组织学和免疫组化检查。
免疫组化分析显示,EDS给药后c-erbAα和c-erbAβ的免疫反应性变化相似。在EDS处理后第1天,LC中c-erbAα和c-erbAβ的免疫反应强度随其数量同时下降。EDS给药7天后,既没有LC,也没有c-erbAα和c-erbAβ免疫反应性。在EDS处理14天后发现了第一批阳性染色的LC,此时检测到了LC祖细胞。在EDS处理21天后,再生LC中最显著的c-erbAα和c-erbAβ免疫染色明显;这与LC祖细胞数量增加及其向成年LC群体的转化相吻合。EDS处理35天后,c-erbAα和c-erbAβ阳性LC数量丰富,它们在睾丸间质中的数量和定位与对照大鼠非常相似。
在EDS处理后LC再填充过程中观察到的c-erbAα和c-erbAβ免疫反应强度变化与祖细胞分化为成熟LC的过程相对应。获得的结果支持甲状腺激素在青春期前大鼠睾丸LC分化中起调节作用的观点。