采用干涉相差显微术和实时荧光定量PCR (Q-PCR)技术显示,腹毛类纤毛虫包囊游仆虫(Euplotes encysticus)包囊形成和解脱过程中,细胞微管胞器始终处于非装配及装配的功能活动状态,脱包囊过程中伸缩泡的运动对细胞充分吸水用于微管胞器的再装配及细胞运动起了关键的作用,细胞内α-、β-和γ-微管蛋白基因的表达量相伴发生变化.其中,微管胞器去分化时,细胞形成毛基体非吸收型包囊,细胞α-、β-和γ-微管蛋白基因的表达量整体呈现逐渐减少的趋势,其细胞微管蛋白基因的相对表达量从大到小依次是β-、α-和γ-微管蛋白基因,但α-、β-微管蛋白基因的起始拷贝数远大于后一种微管蛋白基因的表达量;微管胞器再分化时,伴随着伸缩泡的剧烈伸缩运动,口纤毛器和体纤毛器微管先后形成,细胞在包囊内做旋转运动并脱囊而出,迅速转化为营养细胞,期间细胞内α-、β-和γ-微管蛋白基因的表达量呈现从小到大增加的趋势,其微管蛋白基因的相对表达量从大到小依次是β-、α-和γ-微管蛋白基因,而前两种微管蛋白基因的起始拷贝数远大于后一种微管蛋白基因的表达量.结果表明,包囊游仆虫形成包囊和脱包囊过程中,伴随着皮层微管胞器的不同分化,细胞内α-、β-和γ-微管蛋白基因始终处于不同的功能活动状态,其中作为微管组织中心主要组分的γ-微管蛋白也始终处于不同的功能状态.
The processes of encystment and excystment of the hypotrichous ciliate Euplotes encysticus,were investigated using interference contrast microscopy and real-time quantitative PCR (Q-PCR).Results showed that the microtubular organelles were always in non-assembly and assembly functional states; furthermore,the activity of the contractile vacuole during excystment played an important role in reassembly of cortical microtubular organelles and cellular activities,and meanwhile,the expression level of α-,β-and γ-tubulin genes changed relatively.During the dedifferentiation of the cortical microtubular organelles,cells formed the non-kinetosome-resorbing cysts and the expression of α-,β-and γ-tubulin genes showed a downward trend.During the re-differentiation of the cortical microtubular organelles,oral and somatic ciliature formed in order and cells rotated inside the cyst and soon after excysted to vegetative cells,while the gene expression of α-,β-and γ-tubulin showed an increasing tendency.During the whole process,the gene expression quantity from more to less were β-,α-and γ-tubulin,and the initial copies of the former two were much more than the latter's.It was revealed that during the processes of encystment and excystment of Euplotes encysticus,with the differentiation of the cortical microtubule organelles,α-,β-and γ-tubulin gene were always in the different functional states relatively intracellular,as well as the γ-tubulin gene which works as microtubule organizing center.