Dluzewski A R, Fryer P R, Griffiths S, Wilson R J, Gratzer W B
Medical Research Council Cell Biophysics Unit, King's College, London, UK.
J Cell Sci. 1989 Apr;92 ( Pt 4):691-9. doi: 10.1242/jcs.92.4.691.
Immuno-gold labelling electron microscopy of thin sections was used to determine the distribution of red cell membrane and membrane skeleton proteins in the vicinity of internalized malaria parasites. When examined immediately after invasion (young ring-stage parasites), the parasitophorous vacuole membranes of both Plasmodium falciparum and P. knowlesi were found to be characterized by the essentially complete absence of spectrin, ankyrin and the most abundant transmembrane protein, band 3. P. knowlesi merozoites were trapped in the attached but not internalized state by pretreatment with cytochalasin B. In this merozoite-red cell complex antibody labelling showed that band 3 had been eliminated from the region of the host cell membrane in contact with the parasite. Internal vesicles, originating apparently from the site of attachment, were often observed in the red cell. Opposite the attached parasite a cavity was also sometimes seen in the host cell, presumably representing an incipient internal vesicle. The membrane was intact, as judged by the absence of protein (haemoglobin) in the cavity, and, like the membranes surrounding the internal vesicles, was devoid of membrane proteins. A large multilamellar body was sometimes seen in the merozoite close to its point of attachment. The lamellar spacing was about 50 nm. The electron microscope images suggest a diffusion of electron-dense material from the lamellar body into the cavity in the host cell.
利用薄切片免疫金标记电子显微镜技术来确定内化疟原虫附近红细胞膜和膜骨架蛋白的分布。在入侵后立即检查(年轻环状期疟原虫)时,发现恶性疟原虫和诺氏疟原虫的寄生泡膜的特征是基本上完全没有血影蛋白、锚蛋白以及最丰富的跨膜蛋白带3。用细胞松弛素B预处理后,诺氏疟原虫裂殖子被困在附着但未内化的状态。在这种裂殖子-红细胞复合物中,抗体标记显示与寄生虫接触的宿主细胞膜区域中的带3已被清除。在红细胞中经常观察到明显起源于附着部位的内部囊泡。在附着的寄生虫对面,有时在宿主细胞中也可见一个腔,推测代表一个初始内部囊泡。根据腔内没有蛋白质(血红蛋白)判断,膜是完整的,并且与围绕内部囊泡的膜一样,没有膜蛋白。有时在裂殖子靠近其附着点处可见一个大的多层体。层间距约为50纳米。电子显微镜图像表明电子致密物质从层状体扩散到宿主细胞的腔中。