Biochemical characterization and immunohistochemical identification of vitellin in the ovary of kuruma prawn (Marsupenaeus japonicus)
2004
Tahara, D. (Fukui Prefectural Univ., Obama (Japan)) | Nakano, K. | Miyadai, T. | Yano, I.
Vitellin (Vn) was purified from the mature ovaries of the kuruma prawn (Marsupenaeus japonicus) with gel filtration, Polyacrylamide Gel Electrophoresis (PAGE), and electro-elution. Native- and SDS-PAGE revealed that the purified Vn was a lipo-glyco-caroteno-protein of 530 kDa consisting of four sub units (70, 83, 105, and 190 kDa). Gly, Ala, and Glu were the dominant amino acids of Vn. The antiserum against the purified Vn, which was raised in a rabbit, immuno-reacted with Vitellogenin (VTG) in the hemolymph and Vn in the ovary by immunodiffusion and Western blotting. Positive immuno-reaction with the antiserum was detected in the cytoplasm of oocytes at the late perinucleolus stage. However follicle cells surrounding the oocytes were not immuno-stained at any developmental stage. No immuno-reaction was detected in the cortical rods of mature oocytes. Immuno-histochemical observation revealed that the accumulation of Vn in the cytoplasm of the oocytes began at the early vitellogenic stage. These results indicate that the level of in situ cellular VTG in follicle cells is below the limit of immunodetection.
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