Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
Laboratório de Aquacultura, Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
Mol Reprod Dev. 2019 Nov;86(11):1592-1602. doi: 10.1002/mrd.23245. Epub 2019 Jul 29.
Follicular atresia is a hormonally controlled degenerative process involving apoptosis of the somatic and germ cells. Since different signaling pathways can induce cell death, the aim of the present study was to investigate cell death signaling and crosstalk between autophagic, apoptotic, and lysosomal proteins during follicular atresia in Nile tilapia. For this, females were kept in controlled conditions for 21 days, and ovary samples were collected weekly. The atretic follicles (AF) were analyzed in three regression phases: Early, advanced, and late. Under electron microscopy, the follicular cells exhibited numerous protein synthesis organelles in the early AF. Immunoreactivity for Bcl2, Beclin1, Lc3, and Cathepsin D increased significantly in advanced AF (p < .001), when follicular cells were in intense yolk phagocytosis. In this phase, autophagosomes and autolysosomes were frequently observed. In the late AF, follicular cells had a markedly electron-lucid cytoplasm and immunoreactivity for Bax and TUNEL assay indicated an elevated apoptosis rate. Colocalisation of Lamp1/Cathepsin D and Lc3/Caspase-3 suggests dynamic crosstalk between the autophagy, apoptosis, and lysosome pathways. Taken together, the data indicate that autophagy plays a role in the homeostasis and clearance of the follicular cells preceding Cathepsin D mediated apoptosis during follicular atresia in Nile tilapia.
卵泡闭锁是一个受激素控制的退化过程,涉及体细胞和生殖细胞的凋亡。由于不同的信号通路可以诱导细胞死亡,本研究的目的是研究在尼罗罗非鱼卵泡闭锁过程中细胞死亡信号和自噬、凋亡和溶酶体蛋白之间的串扰。为此,将雌性鱼在受控条件下饲养 21 天,并每周采集卵巢样本。对处于三个退化阶段的闭锁卵泡(AF)进行分析:早期、晚期和晚期。在电子显微镜下,早期 AF 的卵泡细胞显示出许多蛋白质合成细胞器。晚期 AF 中 Bcl2、Beclin1、Lc3 和组织蛋白酶 D 的免疫反应性显著增加(p < .001),此时卵泡细胞正在进行强烈的卵黄吞噬作用。在这个阶段,经常观察到自噬体和自溶体。在晚期 AF 中,卵泡细胞的细胞质明显透亮,Bax 的免疫反应性和 TUNEL 检测表明凋亡率升高。Lamp1/组织蛋白酶 D 和 Lc3/Caspase-3 的共定位表明自噬、凋亡和溶酶体途径之间存在动态串扰。综上所述,数据表明在尼罗罗非鱼卵泡闭锁过程中,自噬在 Cathepsin D 介导的凋亡之前发挥作用,以维持卵泡细胞的稳态和清除。