title: Effects of RNA interference-mediated knock-down of hypoxia-inducible factor-α on respiratory burst activity of the Pacific oyster Crassostrea gigas hemocytes.
abstract: In mammals, hypoxia-inducible factor-1 α (HIF-1α) is known to play important roles not only in oxygen homeostasis but also in innate immune responses. In this study, to assess the functional role of HIF-α in respiratory burst activity of Crassostrea gigas hemocytes, oysters were injected with HIF-α- or green fluorescent protein (GFP)-targeted-long double-stranded RNAs (dsRNAs), and at 1, 3, and 7 days post-injection, knock-down of C. gigas HIF-α expression and production of reactive oxygen species (ROS) were analyzed. Expression of HIF-α in mantle, gill, and hemocytes of C. gigas was clearly down-regulated by injection of the HIF-α-targeted-long dsRNA, but was not inhibited by the GFP-targeted-long dsRNA, indicating that HIF-α expression was suppressed through sequence-specific and systemic RNA interference (RNAi). Respiratory burst activity of hemocytes was significantly increased by administration of GFP-targeted-long dsRNA. However, knock-down of HIF-α expression led to significant decrease of chemiluminescence (CL) response of C. gigas hemocytes at 3 and 7 days post-administration of HIF-α-targeted-long dsRNA, indicating the critical role of HIF-α in activation of respiratory burst activity of oyster hemocytes.
authors.affiliation: Department of Aquatic Life Medicine, Pukyong National University, 599-1, Daeyeondong, Namgu, Busan 608-737, Republic of Korea.
authors.forename: Seung Hyuk
authors.initials: SH
authors.lastname: Choi
authors.forename: Bo Young
authors.initials: BY
authors.lastname: Jee
authors.forename: Su Jin
authors.initials: SJ
authors.lastname: Lee
authors.forename: Mi Young
authors.initials: MY
authors.lastname: Cho
authors.forename: Soon Jeong
authors.initials: SJ
authors.lastname: Lee
authors.forename: Jin Woo
authors.initials: JW
authors.lastname: Kim
authors.forename: Hyun Do
authors.initials: HD
authors.lastname: Jeong
authors.forename: Ki Hong
authors.initials: KH
authors.lastname: Kim
mesh_terms: D000818:Animals; D049896:Crassostrea / Q000235:genetics* / Q000378:metabolism; D055785:Gene Knockdown Techniques; D006434:Hemocytes / Q000378:metabolism; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics* / Q000378:metabolism; D008163:Luminescent Measurements; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000235:genetics* / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism*; D016897:Respiratory Burst
chemical_list: D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D012330:RNA, Double-Stranded; D017382:Reactive Oxygen Species
keywords: Crassostrea gigas; Hypoxia-inducible factor-α; Knock-down of HIF-α; RNA interference; Respiratory burst activity