Institute of Aquaculture, University of Stirling, Stirling, FK9 3LA, Scotland, UK,
Fish Physiol Biochem. 2013 Oct;39(5):1101-14. doi: 10.1007/s10695-012-9767-1. Epub 2013 Jan 11.
This study examines the structural differentiation of the apical crypts of mitochondria-rich cells (MRCs) in Nile tilapia as a response to osmotic challenge. Larvae were transferred from freshwater at 3 days post-hatch to 12.5 and 20 ppt and were sampled at 24- and 48-h post-transfer. Scanning electron microscopy allowed quantification of MRCs, based on apical crypt appearance and surface area, resulting in a morphological classification of 'sub-types', that is, Type I or absorptive (surface area range 5.2-19.6 μm(2)), Type II or active absorptive form (surface area range 1.1-15.7 μm(2)), Type III or weakly functioning form (surface area range 0.08-4.6 μm(2)) and Type IV or active secreting form (surface area range 4.1-11.7 μm(2)). Mucus cell crypts were discriminated from those of MRCs based on the presence of globular extensions and quantified. Density and frequency of MRCs and mucus cells varied significantly according to the experimental salinity and time post-transfer; in freshwater-adapted larvae, all types were present except Type IV but, following transfer to elevated salinities, Type I and Type II disappeared and appeared to be replaced by Type IV crypts. Type III crypt density remained constant following transfer. Transmission electron microscopy with immunogold labelling, using a novel pre-fixation technique with anti-Na(+)/K(+)-ATPase, allowed complementary ultrastructural visualisation of specific localisation of the antibodies on active MRCs, permitting a review of MRC apical morphology and related Na(+)/K(+)-ATPase binding sites.
本研究考察了尼罗罗非鱼顶泌细胞(MRCs)的顶端隐窝结构分化,作为其对渗透压挑战的反应。将孵化后 3 天的幼虫从淡水中转移到 12.5 和 20 ppt,并在转移后 24 和 48 小时取样。扫描电子显微镜允许根据顶泌细胞的隐窝外观和表面积来量化 MRCs,从而对“亚型”进行形态分类,即 I 型或吸收型(表面积范围 5.2-19.6 μm²)、II 型或活跃吸收型(表面积范围 1.1-15.7 μm²)、III 型或功能较弱型(表面积范围 0.08-4.6 μm²)和 IV 型或活跃分泌型(表面积范围 4.1-11.7 μm²)。根据存在球状延伸的特征,可以将粘液细胞隐窝与 MRC 隐窝区分开来,并进行量化。根据实验盐度和转移后时间的不同,MRC 和粘液细胞的密度和频率有显著变化;在适应淡水的幼虫中,除了 IV 型外,所有类型都存在,但转移到高盐度后,I 型和 II 型消失,似乎被 IV 型隐窝取代。转移后,III 型隐窝密度保持不变。使用新型的抗 Na(+)/K(+)-ATPase 预固定技术,结合免疫金标记的透射电子显微镜,允许对特定的 MRC 进行抗体的局部定位进行互补的超微结构可视化,从而可以对 MRC 顶泌细胞形态和相关的 Na(+)/K(+)-ATPase 结合位点进行综述。