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Zhongliang Wang,Bei Wang,Gang Chen,Yishan Lu,Jichang Jian,Zaohe Wu 한국유전학회 2017 Genes & Genomics Vol.39 No.10
MicroRNAs (miRNAs) are a class of noncoding RNA molecules that function as negative regulators of gene expression and play important roles in a wide spectrum of biological processes, including in immune response. However, the physiological regulation function of Pinctada fucata miRNAs, specially their immunomodulation has not been explored yet. Here, two small RNA libraries from hemocytes of P. fucata with or without Vibrio alginolyticus infection were constructed and sequenced using the high-throughput Illumina deep sequencing technology. In total, 11,939,992 and 11,083,327 raw reads, corresponding to 10,993,546 and 9,988,179 clean reads, were respectively obtained in the control and infected libraries. A total of 276 miRNAs, including 225 known miRNAs and 51 putative novel miRNAs, were identified by bioinformatic analysis. By using pairwise comparison between two libraries, 93 miRNAs were found to be significantly differentially expressed, with 42 and 51 miRNAs exhibiting up-regulation and down-regulation, respectively. Thereinto, some known miRNAs were considered to be immune-related. Real-time PCR were implemented for 6 miRNAs co-expressed in the control and infected samples, and agreement was confirmed between the high-throughput sequencing and real-time PCR data. After miRNA targets were predicted, GO and KEGG pathway enrichment analysis were performed, and the results indicated that ten of the differentially expressed miRNAs were involved in immunerelated pathways, and might participate in the host immune response to V. alginolyticus. These results of identification and comparative analysis of miRNAs might deepen our understanding of host-pathogen interactions and immune defense mechanisms in P. fucata.
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Pin Nie,Yishan Lu,Zhen Gan,Bei Wang,Wei Zhou,Yanfei Zhang,JiChang Jian,Zaohe Wu 한국유전학회 2016 Genes & Genomics Vol.38 No.4
The 70 kDa zeta-chain associated protein kinase (ZAP-70) plays a critical role in effective signal transductions that are fundamental to T cell differentiation, proliferation and effector functions. In this paper, the ZAP- 70 gene of Nile tilapia, Oreochromis niloticus (designated as On-ZAP-70) was cloned and its expression pattern under the stimulation of Streptococcus agalactiae was investigated. Sequence analysis showed important structural characteristics required for TCRs signal transduction were detected in the deduced amino acid sequence of On-ZAP- 70, and the deduced genomic structure of On-ZAP-70 was similar to the known ZAP-70. In healthy tilapia, the On- ZAP-70 transcripts were mainly detected in the thymus, spleen, head kidney and gill. Moreover, there was a clear time-dependent expression pattern of On-ZAP-70 after immunization and the expression reached the highest level at 48 h in the spleen and head kidney, and at 72 h in the thymus, respectively. This is the first report on the expression of ZAP-70 induced by intracellular bacteria vaccination in teleosts. These findings indicated that On- ZAP-70 may play an important role in the immune response to intracellular bacteria in Nile tilapia.
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