Chen Shun, Cheng An-Chun, Wang Ming-Shu, Zhou Yi
Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Yaan 625014, Sichuan, China.
Bing Du Xue Bao. 2008 Sep;24(5):396-400.
The characteristics changes of apoptosis of Duck Embryo Fibroblasts (DEF) cells induced by New type gosling viral enteritis virus, NGVEV) were observed by means of HE staining, electron microscopy and Annexin V-FITC/PI fluorescent staining. During 24-48 h post infection (pi), the difference of morphological change between infected DEF cells and the mock infected cells was invisible. At 72 h pi, the nuclear chromatin was getting condensed through HE staining; apoptotic morphological change such as abnormal shape of the nucleus, condensation of the cytoplasm and chromatin were observed under electron microscope; and the early apoptotic cells (Annexin V-FITC positive and PI negative) were detected under fluorescence microscope. At 96-120 h pi, by means of HE staining and electron microscopy, the advanced morphological change of apoptosis such as formation of different kinds of apoptotic bodies, and shrink of the DEF cells and nucleus were detected; under fluorescence microscope the different stages of the apoptotic DEF can be easily distinguished: early apoptotic cells (Annexin V-FITC postive and pi negative), advanced or late apoptotic cells (both Annexin V-FITC and PI positive), necrosis cells or dead cells (Annexin V-FITC negative and PI positive). This investigation shows that NGVEV might induce apoptosis and form characteristic apoptotic morphological changes in the DEF cells. NGVEV inducement of apoptosis may be an important mechanism of efficient dissemination of virus progeny.
通过苏木精-伊红(HE)染色、电子显微镜观察以及膜联蛋白V-异硫氰酸荧光素/碘化丙啶(Annexin V-FITC/PI)荧光染色,观察新型鹅细小病毒(NGVEV)诱导鸭胚成纤维细胞(DEF)凋亡的特征性变化。在感染后24至48小时(pi),感染DEF细胞与 mock感染细胞之间的形态学变化差异不可见。在感染后72小时,通过HE染色可见细胞核染色质浓缩;在电子显微镜下观察到凋亡形态学变化,如细胞核形态异常、细胞质和染色质浓缩;在荧光显微镜下检测到早期凋亡细胞(膜联蛋白V-FITC阳性且PI阴性)。在感染后96至120小时,通过HE染色和电子显微镜观察到凋亡的晚期形态学变化,如形成各种凋亡小体,以及DEF细胞和细胞核缩小;在荧光显微镜下可轻松区分凋亡DEF的不同阶段:早期凋亡细胞(膜联蛋白V-FITC阳性且PI阴性)、晚期凋亡细胞(膜联蛋白V-FITC和PI均阳性)、坏死细胞或死亡细胞(膜联蛋白V-FITC阴性且PI阳性)。本研究表明,NGVEV可能诱导DEF细胞凋亡并形成特征性凋亡形态学变化。NGVEV诱导凋亡可能是病毒子代高效传播的重要机制。