Polyclonal antisera against estuarine crustacean vitellins a molecular approach to reproductive endocrinology and toxicology

Tuberty, S.R.*; Nates, S.F.; McKenney Jr., C.L.: Polyclonal antisera against estuarine crustacean vitellins: a molecular approach to reproductive endocrinology and toxicology.

In order to fully elucidate the action of crustacean hormones, or their agonists, on reproduction and vitellogenesis it has become increasingly important to develop sensitive assays that indicate a stimulatory or inhibitory effect on easily measured endpoints. Because of the relative abundance of vitellin in crustacean yolk and the ease with which it can be isolated, vitellin makes an excellent model for studying the mechanisms that control and regulate reproduction. With this in mind, adult female Lepidophthalmus louisianensis, Palaemonetes pugio, Rhithropanopeus harrisii, Americamysis (Mysidopsis) bahia, and Uca panacea were collected from estuarine localities in Santa Rosa Sound, Gulf Breeze, Florida during late spring and summer of 1999. Vitellin was purified from each crustacean and used to immunize rabbits for polyclonal antibody production. Specificity of each antisera was tested against their respective vitellins by Western blotting. The antisera will be used to develop enzyme-linked immunosorbent assays for determination of hemolymph vitellogenin and egg vitellin concentrations. These assays could be used to determine effects of endogenous hormones on the rate or onset of vitellogenesis. In addition, these assays would be valuable tools in determining the effects of insect juvenile hormone agonists on reproduction and egg maturation.

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