Bonaldo M C, Souto-Padron T, de Souza W, Goldenberg S
Fundacao Oswaldo Cruz (FIOCRUZ), Departamento de Bioquímica e Biologia Molecular, Rio de Janeiro, Brazil.
J Cell Biol. 1988 Apr;106(4):1349-58. doi: 10.1083/jcb.106.4.1349.
The transformation of Trypanosoma cruzi epimastigotes to the mammal infective metacyclic trypomastigotes (metacyclogenesis) can be performed in vitro under chemically defined conditions. Under these conditions, differentiating epimastigotes adhere to a surface before their transformation into metacyclic trypomastigotes. Scanning and transmission electron microscopy of adhered and non-adhered parasites during the metacyclogenesis process show that only epimastigotes and few transition forms are found in the first population, whereas metacyclic trypomastigotes are exclusively found in the cell culture supernatant. PAGE analysis of the [35S]methionine metabolic labeling products of adhered and non-adhered parasites shows that although most of the polypeptides are conserved, adhered parasites express specifically four polypeptides in the range of 45-50 kD with an isoelectric point of 4.8. These proteins might be involved in the adhesion process and are recognized by an antiserum against total adhered parasite proteins. This antiserum also recognized a group of 45-50 kD in the iodine-radiolabeled surface proteins of differentiating cells, providing direct evidence that these components are indeed surface antigens. The results suggest that epimastigotes must adhere to a substrate before their transformation to metacyclic trypomastigotes, being released to the medium as the metacyclogenesis process is accomplished. This could correspond to the process naturally occurring within the triatomine invertebrate host.
克氏锥虫上鞭毛体向具有感染哺乳动物能力的循环后期锥鞭毛体的转变(循环后期发育)可在体外化学限定条件下完成。在这些条件下,正在分化的上鞭毛体在转变为循环后期锥鞭毛体之前会附着于一个表面。对循环后期发育过程中附着和未附着的寄生虫进行扫描电子显微镜和透射电子显微镜观察发现,在第一批群体中仅发现上鞭毛体和少数过渡形态,而循环后期锥鞭毛体仅存在于细胞培养上清液中。对附着和未附着的寄生虫的[35S]甲硫氨酸代谢标记产物进行的PAGE分析表明,尽管大多数多肽是保守的,但附着的寄生虫特异性表达四种分子量在45 - 50 kD范围内、等电点为4.8的多肽。这些蛋白质可能参与附着过程,并能被针对总附着寄生虫蛋白质的抗血清识别。该抗血清还识别正在分化细胞的碘放射性标记表面蛋白中的一组45 - 50 kD的蛋白,这提供了直接证据表明这些成分确实是表面抗原。结果表明,上鞭毛体在转变为循环后期锥鞭毛体之前必须附着于一个底物上,并在循环后期发育过程完成时释放到培养基中。这可能对应于在锥蝽无脊椎动物宿主体内自然发生的过程。