Characterization, expression and localization of valosin-containing protein in ovaries of the giant tiger shrimp Penaeus monodon

摘要

Valosin-containing protein (VCP), a member of the ATPase-associated with diverse cellular activity (AAA) family, was identified from the giant tiger shrimp (Penaeus monodon). The full-length cDNA of the PmVCP mRNA consisted of 2724 bp containing an ORF of 2367 bp corresponding to a deduced polypeptide of 788 amino acids. The deduced PmVCP protein contained two putative Cdc48 domains (positions 17-103, E-value = 2.00e-36 and 120-186, E-value = 3.60e-11) and two putative AAA domains (positions 232-368, E-value =3.67e-24 and 505-644, E-value = 3.73e-25). PmVCP mRNA expression in ovaries was greater than that in testes in both juveniles and broodstock. PmVCP was significantly up-regulated in stages II and IV ovaries in intact wild broodstock (P 0.05). The expression level of PmVCP mRNA in ovaries of 14-month-old shrimp was not affected by progesterone injection (0.1 mu g/g body weight, P> 0.05). In contrast, exogenous 5-HT administration (50 mu g body weight) resulted in an increase of PmVCP mRNA in ovaries of 18-month-old shrimp at 6 and 24 h post-injection (hpi) (P < 0.05). The rPmCdc48-VCP protein and its polyclonal antibody were successfully produced. Cellular localization revealed that PmVCP was localized in the ooplasm of previtellogenic oocytes. Subsequently, it was translocated into the germinal vesicle of vitellogenic oocytes. Interestingly, PmVCP was found in nucleo-cytoplasmic compartments, in the cytoskeletal architecture and in the plasma membrane of mature oocytes in both intact and eyestalk-ablated broodstock. (C) 2013 Elsevier B.V. All rights reserved.