Gallo J M, Ouaissi A, Capron A, Schrével J
Laboratoire de Biologie Cellulaire (UA CNRS No. 290), UFR Sciences Fondamentales et Appliquées, Poitiers, France.
Exp Parasitol. 1987 Oct;64(2):133-8. doi: 10.1016/0014-4894(87)90136-6.
The distribution of tubulin domains in the mammalian stages of Trypanosoma cruzi was investigated by using monoclonal antibodies elicited against bovine brain tubulin. Western blotting performed on T. brucei trypomastigotes and T. cruzi epimastigotes showed that the monoclonal antibodies 16D3 and 24E3 reacted only with tubulin in these cell types. Indirect immunofluorescence revealed that, whereas 16D3 stained all microtubules, including subpellicular microtubules, the epitope defined by 24E3 was found in only a part of the tubulin pool of amastigotes and intermediate stages infecting murine fibroblasts and of broad trypomastigotes; the staining was limited to the basal bodies and the distal region of the flagellar adhesion zone in these developmental forms. By contrast, slender trypomastigotes did not exhibit any reaction with 24E3. These results are consistent with a transformation of broad trypomastigotes into slender trypomastigotes during which the tubulin domain recognized by 24E3 would undergo modifications leading to its complete masking in slender forms. The morphogenesis of the mammalian stages of T. cruzi would involve modifications of the tubulin molecule.
利用针对牛脑微管蛋白产生的单克隆抗体,研究了克氏锥虫哺乳动物阶段微管蛋白结构域的分布情况。对布氏锥虫锥鞭毛体和克氏锥虫上鞭毛体进行的蛋白质免疫印迹分析表明,单克隆抗体16D3和24E3仅与这些细胞类型中的微管蛋白发生反应。间接免疫荧光显示,16D3可对所有微管进行染色,包括表膜下微管,而24E3所定义的表位仅存在于感染小鼠成纤维细胞的无鞭毛体和中间阶段以及粗大锥鞭毛体的部分微管蛋白库中;在这些发育形态中,染色仅限于基体和鞭毛附着区的远端区域。相比之下,细长锥鞭毛体与24E3没有任何反应。这些结果与粗大锥鞭毛体向细长锥鞭毛体的转变一致,在此过程中,24E3识别的微管蛋白结构域会发生修饰,导致其在细长形态中完全被掩盖。克氏锥虫哺乳动物阶段的形态发生可能涉及微管蛋白分子的修饰。