Burge Erin J, Madigan Daniel J, Burnett Louis E, Burnett Karen G
Grice Marine Laboratory, College of Charleston and Hollings Marine Laboratory, 331 Fort Johnson Road, Charleston, SC 29412, USA.
Fish Shellfish Immunol. 2007 Apr;22(4):327-39. doi: 10.1016/j.fsi.2006.06.004. Epub 2006 Jun 15.
The purpose of this study was to quantify the gene expression of lysozyme, an important antibacterial protein produced by shrimp hemocytes, within tissues of Litopenaeus vannamei Boone in response to a pathogen challenge. We quantified lysozyme transcripts with a real-time PCR method and used these data, along with total hemocyte counts, to infer patterns of hemocyte trafficking during the immune response. Transcript expression was detected by in situ hybridization of mRNA in circulating hemocytes, and within tissues with high hemocyte concentrations. Lysozyme gene expression was monitored in 5 tissues and in circulating hemocytes for 48 h following challenge with the shrimp pathogen Vibrio campbellii Baumann. The results suggest that lysozyme is expressed in most if not all hemocytes in circulation and in peripheral tissues. Injection with V. campbellii produced a significant decrease in transcriptional signal in circulating hemocytes and peripheral tissues 4 h after injection. Over the same early time period lysozyme signal increased significantly in the muscle at the site of injection and remained high for the duration of the time-course, suggesting that hemocytes are recruited to the site of injection early during the course of the immune response. After 4 h, lysozyme signal increased in circulating hemocytes and tissues, with a return to control levels noted for all tissues except the muscle at the site of injection.
本研究的目的是量化凡纳滨对虾（Litopenaeus vannamei Boone）组织中溶菌酶（一种由对虾血细胞产生的重要抗菌蛋白）的基因表达，以应对病原体攻击。我们采用实时定量PCR方法对溶菌酶转录本进行定量，并利用这些数据以及总血细胞计数，推断免疫反应期间血细胞的迁移模式。通过对循环血细胞以及血细胞浓度高的组织中的mRNA进行原位杂交来检测转录本表达。在用对虾病原体坎氏弧菌（Vibrio campbellii Baumann）攻击后的48小时内，监测了5种组织和循环血细胞中的溶菌酶基因表达。结果表明，溶菌酶在循环中的大多数（如果不是全部）血细胞以及外周组织中表达。注射坎氏弧菌后，注射后4小时，循环血细胞和外周组织中的转录信号显著降低。在同一早期时间段内，注射部位肌肉中的溶菌酶信号显著增加，并在整个时间进程中保持较高水平，这表明在免疫反应过程早期，血细胞被招募到注射部位。4小时后，循环血细胞和组织中的溶菌酶信号增加，除注射部位的肌肉外，所有组织的溶菌酶信号均恢复到对照水平。
