Olea G B, Aguirre M V, Lombardo D M
Universidad Nacional del Nordeste, Facultad de Medicina, Laboratorio de Investigaciones Bioquímicas y Médicas (LIBIM) - CONICET, Moreno 1240, Corrientes, C.P. 3400, Argentina.
Universidad de Buenos Aires, Facultad de Ciencias Veterinarias, Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Chorroarin 280, Buenos Aires (CABA), C.P. 1428, Argentina.
Acta Histochem. 2018 Jul;120(5):446-455. doi: 10.1016/j.acthis.2018.05.010.
In this work, testicular ontogeny is analyzed at the anatomical, histological and immunohistochemical levels; the latter through the detection of GnRHR and PCNA in the testicles of embryos, neonates and juveniles of Columba livia. We analyzed 150 embryos, 25 neonates and 5 juveniles by means of observations under a stereoscopic magnifying glass and scanning electron microscope (SEM). The histological analysis was performed using hematoxylin-eosin staining techniques and the PAS reaction. For the immunohistochemical analysis, the expression of GnRHR and PCNA in embryos corresponding to stages 41, 43 and in neonates of 2, 5, 7 and 75 days post-hatch was revealed in testicular histological preparations. That gonadal outline is evident in stage 18. In stage 29, the testes are constituted of a medulla in which the PGCs are surrounded by the Sertoli cells, constituting the seminiferous tubules. From stage 37 a greater organization of the tubules is visualized and at the time of hatching the testicle is constituted of the closed seminiferous tubules, formed of the PGCs and Sertoli cells. The Leydig cells are evident outside the tubules. In the juvenile stages, the differentiation of germline cells and the organization of small vessels that irrigate the developing testicle begin to be visible. In the analyzed stages, the immunodetection of the GnRHR receptor and PCNA revealed specific marking in the plasma membrane and in the perinuclear zone for GnRHR and in the nucleus of the germline cells in juvenile testicles for PCNA. These results can be used as a basis for further study of endocrine regulation events during testicular ontogeny in avian species.
在本研究中,从解剖学、组织学和免疫组织化学水平对家鸽睾丸个体发育进行了分析;后者通过检测家鸽胚胎、新生儿和幼鸽睾丸中的GnRHR和PCNA来实现。我们借助立体放大镜和扫描电子显微镜(SEM)对150个胚胎、25个新生儿和5只幼鸽进行了分析。组织学分析采用苏木精-伊红染色技术和PAS反应。对于免疫组织化学分析,在睾丸组织学切片中显示了GnRHR和PCNA在孵化后第41、43阶段胚胎以及2、5、7和75天新生儿中的表达。性腺轮廓在第18阶段明显可见。在第29阶段,睾丸由髓质构成,其中原始生殖细胞被支持细胞包围,形成生精小管。从第37阶段开始,可以看到小管有更明显的组织结构,孵化时睾丸由封闭的生精小管构成,由原始生殖细胞和支持细胞组成。间质细胞在小管外明显可见。在幼鸽阶段,可以开始看到生殖细胞的分化以及为发育中的睾丸供血的小血管的组织结构。在分析的阶段中,GnRHR受体和PCNA的免疫检测显示,在幼鸽睾丸中,GnRHR在质膜和核周区域有特异性标记,而PCNA在生殖细胞的细胞核中有特异性标记。这些结果可作为进一步研究鸟类睾丸个体发育过程中内分泌调节事件的基础。