Characterization of a Hematopoietic Tissue-Specific Gene, LP5, from Litopenaeus vannamei

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簡歷

基本資料

Project title

Code/計畫編號

MOST104-2311-B019-003

Translated Name/計畫中文名

白蝦造血組織專一性表現基因LP5之研究

Project Coordinator/計畫主持人

Jiann-Horng Leu

Funding Organization/主管機關

National Science and Technology Council

Department/Unit

Institute of Marine Biology

Website

https://www.grb.gov.tw/search/planDetail?id=11580345

Year

2015

Start date/計畫起

01-08-2015

Expected Completion/計畫迄

31-07-2016

Bugetid/研究經費

1077千元

ResearchField/研究領域

生物技術(理)
海洋科學

"甲殼類動物的血細胞在免疫防禦上扮演著舉足輕重的角色，因此血細胞數的恆定對維持一定程度的免疫力是必須的，而血細胞數的恆定則端賴造血作用來加以維持。甲殼類動物的造血作用是在造血組織(Hematopoietic tissue, HPT)中進行，在之前的研究計畫，我們利用抑制性扣除雜交法 (Suppression subtractive hybridization, SSH)在HPT 中找到一個HPT 專一性表現基因LP5, 而它也是首次在蝦類發現的新穎基因。LP5 的全長cDNA 序列為3251 bp，其ORF 可轉譯出738 個胺基酸的蛋白質，預測分子量為82kDa。經由胺基酸序列同源性分析，顯示LP5 蛋白與organic anion transporting polypeptides (OATPs；有機陰離子運輸器)家族蛋白具有很高的相似性。為了釐清LP5 基因是否與造血作用有關，我們將LP5 dsRNA 注入蝦中，將LP5 基因靜默，結果發現，施打LP5 dsRNA 後第一天，血細胞總數(total hemocyte count) 迅即下降，並在第三、五、七天都維持在低點；而注射Luciferase dsRNA 蝦的血細胞總數，在第一天下降至低點後，在第三天慢慢增加，而在第五、七天恢復至未注射前的數目。此外，BrdU incorporation 分析顯示，與luciferase dsRNA 注射蝦相比， LP5 dsRNA 注射蝦的BrdU-incorporated 血細胞數目以及BrdU-incorporated HPT 細胞數目都是下降的。這些實驗結果顯示LP5 基因的確是與造血作用有關，而本新提計畫即是以LP5 為主題，希望能夠經由更多的實驗，確認LP5 基因在白蝦造血作用的重要性，進一步對白蝦甚至是甲殼類動物的造血作用有更深入的了解。" "Hemocytes play prominent roles in crustacean innate immune system, actively participating in both the cellular and humoral defenses. Because of their importance in crustacean immunity, maintaining the numbers of circulating hemocytes at a relative constant level is necessary. In crustaceans, a variety of factors decrease the number of circulating hemocytes, such as microbial infections and environmental stresses, and hematopoiesis is the only way to replenish new circulating hemocytes. The crustacean hematopoiesis occurs in the specific, sheet-like hematopoietic tissues (HPT), which is situated on and covers the dorsal and dorsolateral sides of the stomach. In our previous project, by using Suppression subtractive hybridization technique (SSH), we identified a HPT-specific gene, LP5, from the HPT of Litopenaeus vannamei shrimp. LP5, a novel gene firstly identified in L. vannamei, had a full-length cDNA of 3251 bp, with an ORF that encodes a protein consisting of 738 amino acid residues with a calculated molecular mass of 82 kDa. Amino acid sequence homology analysis showed that LP5 protein has high similarity to members of organic anion transporting polypeptides (OATPs). To investigate whether LP5 is involved in hematopoiesis in L. vannamei, we injected LP5 dsRNA into the shrimp and then determined the total hemocyte count (THC) changes after injection. The results showed that THC dropped dramatically at 1 day post-injection (dpi) and then remained at low levels at 3, 5 and 7 dpi. However, for shrimp injected with luciferase dsRNA, even though THC dropped at 1 dpi, it gradually increased at 3 dpi, and then recovered at 5 and 7 dpi to levels before dsRNA injection. Additionally, by BrdU incorporation assay, we found that the number of both BrdU-incorporated hemocytes and BrdU-incorporated HPT cells was all lowered than that in luciferase dsRNA-injected shrimp. Therefore, all the results suggested that LP5 is involved in hematopoiesis. This new proposed project aims to perform more experiments to further characterize the importance of LP5 in L. vannamei hematopoiesis, so that we could have better understanding of the hematopoiesis process in shrimp and even in crustaceans."

Keyword(s)

甲殼類動物；白蝦；造血作用；造血組織；有機陰離子運輸器
白蝦；造血作用；造血組織；有機陰離子運輸器
造血作用；造血組織；有機陰離子運輸器
造血組織；有機陰離子運輸器
有機陰離子運輸器
Litopneaeus vannamei
hematopoiesis
hematopoietic tissue
organic anion transporting polypeptide
OATP

DSpace CRIS

Characterization of a Hematopoietic Tissue-Specific Gene, LP5, from Litopenaeus vannamei

基本資料

Description