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花生凝集素与虹鳟鱼鳃上皮中富含线粒体的细胞亚群结合。

Peanut lectin binds to a subpopulation of mitochondria-rich cells in the rainbow trout gill epithelium.

作者信息

Goss G G, Adamia S, Galvez F

机构信息

Department of Biology Sciences, University of Alberta, Edmonton, Alberta, Canada T6G 2E9.

出版信息

Am J Physiol Regul Integr Comp Physiol. 2001 Nov;281(5):R1718-25. doi: 10.1152/ajpregu.2001.281.5.R1718.

Abstract

Fluorescently labeled peanut lectin agglutinin (PNA-FITC) was used to identify a subtype of mitochondria-rich (MR) cells in the gills of freshwater rainbow trout. In situ binding of PNA-FITC was visualized by inverted fluorescence microscopy and found to bind to cells on the trailing edge of the filament epithelium as demonstrated by differential interference contrast optics. The amount of PNA-FITC binding on the filament epithelium increased with cortisol pretreatment concomitant with an increased chloride cell fractional area as demonstrated by scanning electron microscopy. Dispersed gill cells were isolated by trypsinization and separated using a discontinuous Percoll density gradient. Cells migrating to the 1.06-1.09 g/ml interface were found to be MR as demonstrated by staining with the vital mitochondrial dye 4-(4-(dimethylamino)styryl)-N-methylpyridinium iodide and transmission electron microscopy (TEM). However, only approximately 40% of the MR cells were found to bind PNA-FITC. Cortisol pretreatment increased the relative numbers of MR cells isolated from the dispersed gill cell population, but the relative proportions of PNA binding cells remained unchanged. Ultrastructural analysis of isolated cells in the TEM demonstrated that the MR cell fraction was comprised of a mixed population of chloride cells and pavement cells.

摘要

用荧光标记的花生凝集素(PNA-FITC)来鉴定淡水虹鳟鱼鳃中富含线粒体(MR)细胞的一种亚型。通过倒置荧光显微镜观察PNA-FITC的原位结合情况,并通过微分干涉相差光学显微镜观察发现其与鳃丝上皮后缘的细胞结合。扫描电子显微镜显示,随着皮质醇预处理,鳃丝上皮上PNA-FITC的结合量增加,同时氯细胞的分数面积也增加。通过胰蛋白酶消化分离出分散的鳃细胞,并使用不连续的Percoll密度梯度进行分离。用活性线粒体染料4-(4-(二甲基氨基)苯乙烯基)-N-甲基碘化吡啶染色和透射电子显微镜(TEM)显示,迁移到1.06-1.09 g/ml界面的细胞为MR细胞。然而,仅约40%的MR细胞被发现能结合PNA-FITC。皮质醇预处理增加了从分散的鳃细胞群体中分离出的MR细胞的相对数量,但PNA结合细胞的相对比例保持不变。透射电子显微镜对分离细胞的超微结构分析表明,MR细胞部分由氯细胞和平铺细胞的混合群体组成。

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