Effects of RNA interference-mediated knock-down of hypoxia-inducible factor-α on respiratory burst activity of the Pacific oyster Crassostrea gigas hemocytes

Choi, Seung Hyuk; Jee, Bo Young; Lee, Su Jin; Cho, Mi Young; Lee, Soon Jeong; Kim, Jin Woo; Jeong, Hyun Do; Kim, Ki Hong

doi:10.1016/j.fsi.2013.05.001

Cited by 25 publications

(19 citation statements)

References 47 publications

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“…Silencing of ChTβ4 in oyster was carried out by injection with dsRNA as previously described (Choi et al, 2013). The dsRNA sequences for ChTβ4 and EGFP (dsChTβ4 and dsEGFP, respectively) were prepared with T7 RiboMAX Express RNAi System (Promega, USA) as manufacturer's instructions.…”

Section: Silencing Of Chtβ4 and Circulating Hemocytes Countmentioning

confidence: 99%

A thymosin beta-4 is involved in production of hemocytes and immune defense of Hong Kong oyster, Crassostrea hongkongensis

Zhang

Liu

et al. 2016

Developmental & Comparative Immunology

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Thymosin beta-4 (Tβ4) is a ubiquitous protein with multiple and diverse intracellular and extracellular functions in vertebrates. In this study, the full-length cDNA of Tβ4 was cloned and identified in Crassostrea hongkongensis, designated as ChTβ4. The full-length cDNA of ChTβ4 consists of 530 bp with an open reading frame of 126 bp encoding a 41 amino acid polypeptide. SMART analysis indicated that there is one thymosin domain and a highly conserved actin-binding motif (18LKKTET23) in ChTβ4. In vivo injection of recombinant ChTβ4 protein could significantly increase total hemocytes count in oysters, and knockdown of the expression of ChTβ4 resulted in a significant decrease in the circulating hemocytes. Tissue distribution analysis revealed a ubiquitous presence of ChTβ4, with the highest expression in hemocytes. The upregulated transcripts of ChTβ4 in response to bacterial challenge and tissue injury suggest that ChTβ4 is involved in both innate immunity against pathogen infection and wound healing. Moreover, bacteria-clearance experiment showed ChTβ4 could facilitate the clearance of injected bacteria in oysters. In vivo injection with ChTβ4 resulted in reduction of the intracellular ROS in hemocytes, which was associated with increased expression of antioxidant enzymes Cu/Zn superoxide dismutase (SOD), Catalase, and Glutathione Peroxidase (GPX) by pre-treatment with ChTβ4. These results suggest that ChTβ4 is a thymosin beta-4 homolog and plays a vital role in the immune defense of C. hongkongensis.

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Section: Silencing Of Chtβ4 and Circulating Hemocytes Countmentioning

confidence: 99%

A thymosin beta-4 is involved in production of hemocytes and immune defense of Hong Kong oyster, Crassostrea hongkongensis

Zhang

Liu

et al. 2016

Developmental & Comparative Immunology

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“…Finally, direct investigations using RNA interference (RNAi) experiments were conducted. RNAi uses synthetic double-stranded (ds) RNA to disrupt the translation of target genes (Agrawal et al, 2003;Zamore, 2001) and has been successfully used to assess gene function in several invertebrate taxa, including mollusks (Choi et al, 2013;Fabioux et al, 2009;Huvet et al, 2015). Here, several Dicer-substrate small interfering RNAs (DsiRNAs) were synthesized and used to knockdown the C-type lectins before assessing changes in food selection among silenced oysters.…”

Section: Introductionmentioning

confidence: 99%

Reverse genetics demonstrate the role of mucosal C-type lectins in food particle selection in the oyster Crassostrea virginica

Espinosa

Allam

2018

Journal of Experimental Biology

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Prey selection governs species interactions and regulates physiological energetics of individuals and populations. Suspensionfeeding bivalves represent key species in coastal and estuarine systems for their ecological and economic value. These animals are able to sort and selectively ingest nutritious microalgae from dilute and composite mixtures of particulate matter. This aptitude was suggested to be mediated by interactions between carbohydrates associated with the surface of microalgae and C-type lectins present in mucus covering the feeding organs, although a direct, unequivocal, role of lectins in food sorting in bivalves remains elusive. This study was designed to identify and characterize mucosal C-type lectins from oysters and manipulate the expression of these proteins in order to obtain decisive information regarding their involvement in food choice. Thus, two mucosal C-type lectins (CvML3912 and CvML3914) were identified based on transcriptomic and proteomic information. Transcripts of these lectins were detected in the feeding organs and their expression was upregulated following starvation. Recombinant lectin (rCvML3912) competitively inhibited the binding of commercial mannose/glucose-specific lectins to microalgae. Short Dicersubstrate small interfering RNA (DsiRNA) targeting these two lectins were designed and used to evaluate the effect of gene silencing on food particle sorting. As a result, the abundance of the two cognate transcripts significantly decreased and food sorting ability was significantly reduced among silenced oysters as compared with control animals. Overall, these findings propose a novel concept establishing the role of carbohydrate-protein interactions to provide efficient food particle sorting, and establish a new dimension for the role of evolutionarily conserved mannose/glucose-binding proteins in metazoans.

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“…Hypoxia-inducible factors (HIFs) are implicated to play essential roles in oxygen homeostasis and innate immune responses of various tissues (Choi et al, 2013;Semenza and Wang, 1992). They are heterodimers consisting of two subunits, HIF-a and HIF-b (also known as aryl-hydrocarbon receptor nuclear translocator, ARNT) which belong to the basic helix-loop-helix (bHLH)-Per-Arnt-Sim (PAS) family of transcription factors (Sogawa and Fujii-Kuriyama, 1997;Wang and Zhang, 1995;Wenger et al, 1997).…”

Section: Introductionmentioning

confidence: 99%

HIFs genes expression and hematology indices responses to different oxygen treatments in an ovoviviparous teleost species Sebastes schlegelii

Wen

et al. 2015

Marine Environmental Research

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Effects of RNA interference-mediated knock-down of hypoxia-inducible factor-α on respiratory burst activity of the Pacific oyster Crassostrea gigas hemocytes

Cited by 25 publications

References 47 publications

A thymosin beta-4 is involved in production of hemocytes and immune defense of Hong Kong oyster, Crassostrea hongkongensis

A thymosin beta-4 is involved in production of hemocytes and immune defense of Hong Kong oyster, Crassostrea hongkongensis

Reverse genetics demonstrate the role of mucosal C-type lectins in food particle selection in the oyster Crassostrea virginica

HIFs genes expression and hematology indices responses to different oxygen treatments in an ovoviviparous teleost species Sebastes schlegelii

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