Vitellogenins of Oreochromis niloticus: identification, isolation, and biochemical and immunochemical...

Two native forms of vitellogenin (EIP1 and EIP2) were identified in the plasma of Oreochromis niloticus. They were present in females and were estrogen-inducible in males. Both were phosphoglycolipoproteins and both immunoreacted with the antiserum raised against egg proteins. Two prominent bands (EIpp1 and EIpp2; corresponding to 185 and 120 kDa, respectively), observed on sodium dodecyl sulfate-polyacrylamide gels, were induced by estradiol treatment of males and immunoreacted with the antiserum against egg proteins. EIP1 and EIP2 were isolated by precipitation with Mg2+ ions and ethylenediamine tetraacetic acid, preparative polyacrylamide gel electrophoresis, and electroelution. During purification the fractions containing EIP1 and EIP2 also retained EIpp2 and EIpp1, respectively. Immunoblot analyses using affinity-purified antibodies against EIP1, EIP2, EIpp1, and EIpp2 confirmed that EIP1 and EIP2 were at least composed of EIpp2 and EIpp1, respectively. These results suggest that at least two immunochemically different proteins are induced and are secreted into the blood to serve as vitellogenin and to provide a source of nutrient for the developing embryo of Oreochromis niloticus.