Evidence for cell apoptosis suppressing white spot syndrome virus replication in Procambarus clarkii at high temperature.
Evidence for cell apoptosis suppressing white spot syndrome virus replication in Procambarus clarkii at high temperature.
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DOI：
10.3354/dao02532
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发表时间：
2012-12
影响因子：
1.4
通讯作者：
Xiao-Guo Wu;Haitao Xiong;Yi-zhen Wang;H. Du
中科院分区：
文献类型：
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作者：
Xiao-Guo Wu;Haitao Xiong;Yi-zhen Wang;H. Du
在对虾中，较高的水温(~32℃)可以抑制对虾白斑综合征病毒(WSSV)的复制并导致死亡，但其机制尚不清楚。为探讨WSSV感染克氏原虾是否与细胞凋亡有关，采用TdT介导的dUTP缺口末端标记(TUNEL)方法检测了感染WSSV的克氏原虾肝胰腺和鳃细胞中染色体DNA的断裂水平，并将其维持在32℃或24℃±1℃。根据黄绿色TUNEL阳性核的相对细胞数，感染WSSV的克氏原虾在32℃时的细胞凋亡指数明显升高。流式细胞仪对Annexin染色的细胞进行分类显示，颗粒血细胞中最常见的是凋亡，而caspase-3活性的检测显示，它在肝胰腺组织中最高。尽管有这些细胞凋亡的指标，但与WSSV复制在高温下受到限制一致，RT-PCR检测到保持在32℃的对虾中病毒抗凋亡基因ORF390和ORF222的转录没有增加，这可能是由于细胞凋亡水平增加所致。
In shrimp, higher water temperatures (~32°C) can suppress the ability of white spot syndrome virus (WSSV) to replicate and cause mortality, but the mechanisms remain unclear. To investigate whether cell apoptosis might be involved, a Tdt-mediated dUTP nick-end label (TUNEL) method was used to assess levels of chromosomal DNA fragmentation in hepatopancreas and gill cells of Procambarus clarkii crayfish infected with WSSV and maintained at either 32 ± 1°C or 24 ± 1°C. Based on relative cell numbers with yellow-green colored TUNEL-positive nuclei, the apoptotic index was elevated in WSSV-infected crayfish maintained at 32°C. In gill tissue sections examined by transmission electron microscope, cells with nuclei displaying apoptotic bodies or marginated, condensed and fragmented chromatin without concurrent cell cytoplasm damage were also more prevalent. Flow cytometry sorting of annexin-stained cells showed apoptosis to be most prevalent in granular haemocytes, and assays for caspase-3 activity showed it to be most elevated in hepatopancreas tissue. Despite these indicators of cell apoptosis but consistent with WSSV replication being restricted at elevated temperatures, no increases in transcription of the viral anti-apoptosis genes ORF390 and ORF222 were detected by RT-PCR in shrimp maintained at 32°C, possibly due to the elevated levels of cellular apoptosis.