Characterization and expression studies of Gaduscidin-1 and Gaduscidin-2; paralogous antimicrobial peptide-like transcripts from Atlantic cod (Gadus morhua)
Characterization and expression studies of Gaduscidin-1 and Gaduscidin-2; paralogous antimicrobial peptide-like transcripts from Atlantic cod (Gadus morhua)
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DOI：
10.1016/j.dci.2010.11.010
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发表时间：
2011-03-01
影响因子：
2.9
通讯作者：
Rise, Matthew L.
中科院分区：
文献类型：
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作者：
Browne, Mitchell J.;Feng, Charles Y.;Rise, Matthew L.
鱼杀菌素是鱼类抗菌肽的一个家族，是鱼类先天免疫系统的重要组成部分。基于先前生成的大西洋鳕鱼（Cadets morhua）表达序列标签（ESTs）。我们鉴定了代表两种旁系同源AMP样转录物的序列。这些大西洋鳕鱼的旁系同源物被称为gaduscidins（GAD-1和GAD-2），来源于Gadus属的名称。我们使用双向cDNA末端快速扩增（RACE）获得了这些推定AMP编码转录本的全长cDNA序列。GAD-1和GAD-2推定肽显示出与硬骨鱼的piscidin家族成员的序列相似性。在GAD-1和GAD-2的转录表达研究中使用定量逆转录聚合酶链反应（QPCR）。我们研究了这些转录本的组成性表达在六个组织（头肾，血液，脑，鳃，幽门盲囊，脾脏）的非应激少年鳕鱼; GAD-1和GAD-2的转录本检测在所有六个组织中，与最高的表达这两个转录本是在脾脏，头肾，鳃。还在腹膜内（IP）注射福尔马林杀死的非典型杀卫星气单胞菌（ASAL）或磷酸盐缓冲盐水（PBS对照）后，在免疫组织（脾和头肾）中检查GAD-1和GAD-2的转录物表达。这两种转录物在脾脏中被ASAL微弱（小于4倍）上调，但在头肾中对ASAL无反应。由于GAD-1和GAD-2转录本在免疫相关组织（如脾脏和头肾）中组成型高表达，并且在IP注射细菌抗原后在脾脏中诱导较弱，因此它们可能代表大西洋鳕鱼先天免疫的重要组成部分。(C)2010爱思唯尔有限公司版权所有。
Piscidins are a family of antimicrobial peptides (AMPs) from fish that constitute an important component of their innate immune system. Based on previously generated Atlantic cod (Cadets morhua) expressed sequence tags (ESTs). we identified sequences representing two paralogous AMP-like transcripts. These Atlantic cod paralogues were termed gaduscidins (GAD-1 and GAD-2), derived from the genus name Gadus. We obtained full-length cDNA sequences for these putative AMP-encoding transcripts using bi-directional rapid amplification of cDNA ends (RACE). GAD-1 and GAD-2 putative peptides exhibit sequence similarity with members of the piscidin family from teleost fish. Quantitative reverse transcription polymerase chain reaction (QPCR) was utilized in transcript expression studies of GAD-1 and GAD-2. We examined the constitutive expression of these transcripts in six tissues (head kidney, blood, brain, gill, pyloric caecum, and spleen) of non-stressed juvenile cod; GAD-1 and GAD-2 transcripts were detected in all six tissues, with the highest expression of both transcripts being in spleen, head kidney, and gill. Transcript expression of GAD-1 and GAD-2 was also examined in immune tissues (spleen and head kidney) following intraperitoneal (IP) injection of formalin-killed, atypical Aeromonas satmonicida (ASAL) or phosphate-buffered saline (PBS control). Both transcripts were weakly (less than 4-fold) up-regulated by ASAL in spleen but non-responsive to ASAL in head kidney. Since GAD-1 and GAD-2 transcripts are highly expressed constitutively in immune-relevant tissues (e.g. spleen and head kidney), and are weakly induced in spleen following IP injection with bacterial antigens, they may represent important components of innate immunity in Atlantic cod. (C) 2010 Elsevier Ltd. All rights reserved.