Díaz de Barboza Gabriela, Rodríguez Valeria, Ponce Rubén, Theiler Gerardo, Maldonado Cristina, Tolosa de Talamoni Nori
Laboratorio "Cañas", Bioquímica y Biología Molecular, Facultad de Ciencias Médicas, INICSA (CONICET-Universidad Nacional de Córdoba), Córdoba, Argentina.
Química Biológica "A", Facultad de Ciencias Médicas, INICSA (CONICET-Universidad Nacional de Córdoba), Córdoba, Argentina.
Acta Histochem. 2014 Jul;116(6):1037-45. doi: 10.1016/j.acthis.2014.04.007. Epub 2014 Jun 2.
Spermatogenesis is disrupted in Graomys griseoflavus×Graomys centralis male hybrids. This study was aimed to determine whether morphological alterations in Leydig cells from hybrids accompany the arrest of spermatogenesis and cell death of germ cells and whether apoptotic pathways are also involved in the response of these interstitial cells. We used three groups of 1-, 2- and 3-month-old male animals: (1) G. centralis, (2) G. griseoflavus and (3) hybrids obtained by crossing G. griseoflavus females with G. centralis males. Testicular ultrastructure was analyzed by transmission electron microscopy. TUNEL was studied using an in situ cell death detection kit and the expression of apoptotic molecules by immunohistochemistry. The data confirmed arrest of spermatogenesis and intense apoptotic processes of germ cells in hybrids. These animals also showed ultrastructural alterations in the Leydig cells. Fas, FasL and calbindin D28k overexpression without an increase in DNA fragmentation was detected in the Leydig cells from hybrids. In conclusion, the sterility of Graomys hybrids occurs with ultrastructural changes in germ and Leydig cells. The enhancement of Fas and FasL is not associated with cell death in the Leydig cells. Probably the apoptosis in these interstitial cells is inhibited by the high expression of the antiapoptotic molecule calbindin D28k.
黄褐稻鼠×中央稻鼠雄性杂种的精子发生受到破坏。本研究旨在确定杂种莱迪希细胞的形态改变是否伴随着精子发生的停滞和生殖细胞的死亡，以及凋亡途径是否也参与这些间质细胞的反应。我们使用了三组1个月、2个月和3个月大的雄性动物：（1）中央稻鼠，（2）黄褐稻鼠，（3）黄褐稻鼠雌性与中央稻鼠雄性杂交获得的杂种。通过透射电子显微镜分析睾丸超微结构。使用原位细胞死亡检测试剂盒研究TUNEL，并通过免疫组织化学研究凋亡分子的表达。数据证实杂种中精子发生停滞和生殖细胞强烈的凋亡过程。这些动物的莱迪希细胞也表现出超微结构改变。在杂种的莱迪希细胞中检测到Fas、FasL和钙结合蛋白D28k的过表达，而DNA片段化没有增加。总之，稻鼠杂种的不育伴随着生殖细胞和莱迪希细胞的超微结构变化。Fas和FasL的增强与莱迪希细胞的死亡无关。可能这些间质细胞中的凋亡受到抗凋亡分子钙结合蛋白D28k高表达的抑制。
