Molecular characterization of a novel tetraspanin from the oyster, Crassostrea ariakensis: variation, localization and relationship to oyster host defense.
We identified a tetraspanin family member gene, named Ca-TSP, in the oyster Crassostrea ariakensis and found that the transcription profiles of Ca-TSP were variable in the oyster hemocytes. Three distinct patterns of variation of Ca-TSP were observed. Using immunofluorescence and immunoelectron microscopy, we show that Ca-TSP was present in granules and in vesicular structures of the oyster hemocyte. Sequence analysis, structural features and immunogold electron microscopy showed that Ca-TSP is an integral membrane glycoprotein of granules of hemocyte and may be a novel CD63-like gene of the tetraspanin family of molluscs. The gene expression analysis of Ca-TSP using isolated oyster hemocytes, was done following challenge of the oysters with LPS and Poly I:C. The Ca-TSP mRNA levels increased in hemocytes in the first 12 h after LPS and Poly I:C stimulation, and decreased after the addition of H(2)O(2). Western blot analysis using anti-Ca-TSP antibody indicated that gene expression and protein levels were similar. The recombinant Ca-TSP was found to significantly inhibit hemocytes aggregation. Our results suggested that Ca-TSP participates in the innate immunity of the oyster.
Laboratory of Marine Life Science and Technology, College of Animal Sciences, Zhejiang University, Hangzhou, PR China.
Ming
M
Luo
Shigen
S
Ye
Ting
T
Xu
Xinzhong
X
Wu
Ping
P
Yang
D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D049896:Crassostrea / Q000145:classification / Q000187:drug effects / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D006434:Hemocytes / Q000187:drug effects; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007113:Immunity, Innate; D008969:Molecular Sequence Data; D010802:Phylogeny; D011070:Poly I-C / Q000494:pharmacology; D011994:Recombinant Proteins / Q000378:metabolism; D016415:Sequence Alignment; D059470:Tetraspanins / Q000235:genetics* / Q000378:metabolism*
D000276:Adjuvants, Immunologic; D011994:Recombinant Proteins; D059470:Tetraspanins; D006861:Hydrogen Peroxide; D011070:Poly I-C