Chen Shun, Cheng Anchun, Wang Mingshu, Zhu Dekang, Jia Renyong, Luo Qihui, Cui Hengmin, Zhou Yi, Wang Yin, Xu Zhiwen, Chen Zhengli, Chen Xiaoyue, Wang Xiaoyu
Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Yaan 625014, China.
J Vet Sci. 2011 Dec;12(4):333-9. doi: 10.4142/jvs.2011.12.4.333.
In this study, apoptosis was induced by new type gosling viral enteritis virus (NGVEV) in experimentally infected goslings is reported in detail for the first time. After 3-day-old goslings were orally inoculated with a NGVEV-CN strain suspension, the time course of NGVEV effects on apoptotic morphological changes of the internal tissues was evaluated. These changes were observed by histological analysis with light microscopy and ultrastructural analysis with transmission electron microscopy. DNA fragmentation was assessed with a terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay and DNA ladder analysis. A series of characteristic apoptotic morphological changes including chromatin condensation and margination, cytoplasmic shrinkage, plasma membrane blebbing, and formation of apoptotic bodies were noted. Apoptosis was readily observed in the lymphoid and gastrointestinal organs, and sporadically occurred in other organs after 3 days post-infection (PI). The presence and quantity of TUNEL-positive cells increased with infection time until 9 days PI. DNA extracted from the NGVEV-infected gosling cells displayed characteristic 180~200 bp ladders. Apoptotic cells were ubiquitously distributed, especially among lymphocytes, macrophages, monocytes, and epithelial and intestinal cells. Necrosis was subsequently detected during the late NGVEV-infection phase, which was characterized by cell swelling, plasma membrane collapse, and rapidly lysis. Our results suggested that apoptosis may play an important role in the pathogenesis of NGVE disease.
本研究首次详细报道了新型雏鹅病毒性肠炎病毒(NGVEV)在实验感染雏鹅中诱导细胞凋亡的情况。将3日龄雏鹅口服接种NGVEV-CN株悬液后,评估了NGVEV对内脏组织凋亡形态变化的影响时间进程。通过光学显微镜组织学分析和透射电子显微镜超微结构分析观察这些变化。用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(TUNEL)法和DNA梯状分析评估DNA片段化。观察到一系列典型的凋亡形态变化,包括染色质浓缩和边缘化、细胞质收缩、质膜起泡以及凋亡小体形成。感染后3天(PI),在淋巴和胃肠器官中很容易观察到凋亡,在其他器官中偶尔出现。TUNEL阳性细胞的存在和数量随感染时间增加,直至感染后9天。从感染NGVEV的雏鹅细胞中提取的DNA呈现出特征性的180~200 bp梯状条带。凋亡细胞广泛分布,尤其在淋巴细胞、巨噬细胞、单核细胞以及上皮和肠细胞中。在NGVEV感染后期随后检测到坏死,其特征为细胞肿胀、质膜塌陷和迅速溶解。我们的结果表明,凋亡可能在NGVE疾病的发病机制中起重要作用。