Hejmej Anna, Kopera Ilona, Kotula-Balak Malgorzata, Gizejewski Zygmunt, Bilińska Barbara
Department of Endocrinology and Tissue Culture, Institute of Zoology, Jagiellonian University, Krakow, Poland.
J Exp Zool A Ecol Genet Physiol. 2009 Nov 1;311(9):667-75. doi: 10.1002/jez.554.
Morphology and the expression of connexin43 (Cx43) was investigated in testes of immature, prepubertal, and adult European bison bulls by means of routine histology, immunohistochemistry, and Western blotting, respectively. Testes were collected from culled animals living in Bialowieza and Borecka Forests, Poland. Histological examination of testicular tissue of immature and prepubertal males revealed normal structure, whereas of adult individuals either normal testicular structure with advanced spermatogenesis or varying degrees of tubule and interstitial tissue abnormality were seen. Immunohistochemical studies revealed Cx43 signal mostly at Leydig cell membrane appositions. In testes of immature males heterogeneous staining was observed; its intensity markedly increased in prepubertal males reaching almost two times more intense staining in adults. Strong Cx43 signal between Leydig cells was also observed in testes with structural alterations; however, sporadically it was of weak linear pattern. In the tubules of the latter, the intensity of Cx43 staining was weak to moderate or it was even absent. Immunohistochemical qualitative analysis was confirmed by quantitative image analysis in which the staining intensity was expressed as relative optical density of diaminobenzidine deposits. Data from Western blot analyses confirmed the results obtained by immunohistochemistry; immunodetectable Cx43 protein as a band of 43 kDa was detected in all testes samples. Overall, the increase in Cx43 level in testes along the bison postnatal development may be capable of a better exchange of metabolites and coordinating Leydig cell activity during maturation. A relationship between homozygosity occurring in European bison and altered intercellular communication is suggested.
分别通过常规组织学、免疫组织化学和蛋白质免疫印迹法，对未成熟、青春期前和成年欧洲野牛公牛的睾丸进行了形态学及连接蛋白43（Cx43）表达的研究。睾丸取自波兰比亚沃维耶扎和博热茨卡森林中被扑杀的动物。未成熟和青春期前雄性动物睾丸组织的组织学检查显示结构正常，而成年个体的睾丸结构正常且精子发生进展良好，或可见不同程度的小管和间质组织异常。免疫组织化学研究显示，Cx43信号主要出现在睾丸间质细胞膜的相邻处。在未成熟雄性动物的睾丸中观察到染色不均一；其强度在青春期前雄性动物中显著增加，在成年动物中染色强度几乎增加了两倍。在结构改变的睾丸中，也观察到睾丸间质细胞之间有强烈的Cx43信号；然而，偶尔呈弱线性模式。在后者的小管中，Cx43染色强度弱至中等，甚至缺失。免疫组织化学定性分析通过定量图像分析得到证实，其中染色强度以二氨基联苯胺沉积物的相对光密度表示。蛋白质免疫印迹分析的数据证实了免疫组织化学获得的结果；在所有睾丸样本中均检测到免疫可检测的43 kDa条带的Cx43蛋白。总体而言，随着野牛出生后的发育，睾丸中Cx43水平的增加可能有助于更好地进行代谢物交换，并在成熟过程中协调睾丸间质细胞活性。提示欧洲野牛中出现的纯合性与细胞间通讯改变之间存在关联。
