Identification of the Cg Wnt5b gene and its function in melanin production in the Pacific oyster Crassostrea gigas
The Wnt gene family encodes a class of conserved secretory glycoproteins that are widely involved in different biological processes,including cell differentiation and proliferation,cell motility,apoptosis,immune regulation,and pigmentation.In this study,we identified a Wnt family gene,CgWnt5b,and compared the expression of CgWnt5b in black and white shelled oysters in the Pacific oyster Crassostrea gigas.A preliminary study on the function of CgWnt5b in regulating melanogenesis was also conducted.Our results reveal that the CgWnt5b coding sequence is 1 116 bp in length and encodes 371 amino acids.Bioinformatics analysis predicted a relative molecular weight of 42 KDa and an isoelectric point of 9.28 for the CgWnt5b protein.Phylogenetic analysis of CgWnt5b with 19 members of the mammalian Wnt gene family revealed that Cg Wnt5b is a homolog of mammalian Wnt5a and Wnt5b.Quantitative real-time PCR was performed to detect the mRNA distribution of Cg Wnt5b in the mantle,gill,digestive gland,adductor muscle,gonad,labial papls,and hemolymph tissues of the oyster.The results reveal that CgWnt5b is highly expressed in mantle,adductor muscle and labial papls tissues,and has a relatively low expression level in digestive gland,gills,hemolymph,and gonad tissues.The expression of CgWnt5b mRNA in black-shelled oyster was significantly lower than that in white-shelled oyster,and the expression of several melanin-related tyrosinase genes was significantly up-regulated after knockdown of CgWnt5b by RNA interference.In summary,this study identified a member of the Wnt family,CgWnt5b,which may inhibit melanogenesis in C.gigas by suppressing the expression of TYRs,the key factors controlling melanin production.
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