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    ELISA detection of vitellogenin in the haemolymph of Penaeus monodon (Fabricius, 1798) (Caridea: Penaeidae): indirect indicator of female maturity

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    Date
    2017
    Author
    Sukumaran, V.
    Jose, S.
    Chekidhenkuzhiyil, J.
    Lalithabai, M.D.
    Nair, S.B.
    Christo, D.
    Kizhakedath, V.K.
    Paran, B.C.
    Philip, R.
    Sarojini, B.S.I.
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    Abstract
    Developing a non-lethal method for analysis of female reproductive maturity will be more accurate and less invasive than assessing maturity through gonad biopsy. Quantification of the expression levels of vitellogenin (Vg) in haemolymph is a vital tool to assess gonadal maturation as Vg is an indirect indicator of maturity. The lack of commercial antibody/ antisera in India for detection of vitellogenin/vitellin (Vg/Vn) levels in haemolypmh of Penaeus monodon (Fabricius, 1798) lead to the present study. To develop an assay, Vg/Vn subunits isolated together and purified from ovary of mature female P. monodon was used as antigen to raise polyclonal antisera in rabbit. This polyclonal antiserum (anti-PmVg/Vn) was used for developing Vg/Vn enzyme-linked immunosorbent assay (ELISA). Anti-PmVg/Vn demonstrated high specificity in detecting vitellogenin subunits present in haemolymph, vitellin subunits in ovary extract through Western blot analysis, and vitellin-like globules in oocytes by immunostaining/immunohistochemistry. The Vg/Vn ELISA developed through was validated by detecting haemolymph Vg based on gonadosomatic index of immature (stage 1) and sexually mature (stage 2) individuals of P. monodon. The specificity of the anti-PmVg/Vn raised was confirmed by detection of recombinant 74 kDa Vg subunit protein of P. monodon. The expressed recombinant Vg was also used to validate the ELISA. The results have demonstrated the specificity and sensitivity of anti-PmVg/Vn polyclonal antisera; an efficient means to monitor and transform the technology of induced breeding.
    URI
    https://doi.org/10.1093/jcbiol/rux040
    http://drs.nio.org/drs/handle/2264/7834
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