Departamento de Biología, Facultad de Ciencias del Mar y Ambientales, Universidad de Cádiz, Puerto Real, Spain.
J Fish Biol. 2021 Sep;99(3):765-772. doi: 10.1111/jfb.14756. Epub 2021 May 11.
In this study, the authors examined late atretic follicles in the ovaries of Atlantic bluefin tuna, Thunnus thynnus (Linnaeus 1758), at different times of the year using transmitted light and epifluorescence microscopy. Atresia (degeneration and resorption of developing ovarian follicles) is a natural process involved in fecundity downregulation in teleosts and is substantially enhanced in stressful conditions. Early (α and β) atretic stages of yolked oocytes have a relatively short duration in seasonally reproducing species, whereas later (γ and δ) atretic follicles (LAF) persist for longer time in the ovary, serving as a sign of previous vitellogenic activity. LAF can thus be used as reliable markers of maturity during non-reproductive periods. Lipofuscin granules accumulate in the cytoplasm of LAF cells as a result of lysosomal digestion of oocyte components. Taking advantage of the well-known autofluorescent properties of lipofuscins, LAF may be identified in unstained histological sections under fluorescence microscopy using appropriate excitation and emission wavelengths. The authors explore in this study the applicability of fluorescence microscopy to provide a fast and effective method to assess late atresia in fishes. This method may be particularly useful to determine sexual maturity in individuals sampled long after the spawning season, where LAF are difficult to detect in standard histological sections. Furthermore, LAF autofluorescence provides a rapid way to quantify late atresia in fishes using image analysis.
在这项研究中,作者使用透射光和荧光显微镜检查了大西洋金枪鱼(Thunnus thynnus)卵巢中不同时期的晚期闭锁卵泡。闭锁(发育中的卵巢卵泡的退化和吸收)是硬骨鱼类中繁殖力下调的自然过程,在应激条件下会显著增强。卵黄期卵母细胞的早期(α 和 β)闭锁阶段在季节性繁殖物种中的持续时间相对较短,而晚期(γ 和 δ)闭锁卵泡(LAF)在卵巢中持续时间更长,是先前卵黄生成活性的标志。因此,LAF 可作为非繁殖期成熟的可靠标志物。脂褐素颗粒由于溶酶体消化卵母细胞成分而在 LAF 细胞的细胞质中积累。利用脂褐素众所周知的自发荧光特性,在荧光显微镜下使用适当的激发和发射波长,可以在未染色的组织切片中识别 LAF。作者在本研究中探索了荧光显微镜在提供快速有效的方法来评估鱼类晚期闭锁中的适用性。这种方法在产卵季节后很长时间采集的个体中确定性成熟时特别有用,在标准组织切片中很难检测到 LAF。此外,LAF 的自发荧光为使用图像分析定量鱼类晚期闭锁提供了一种快速方法。