Tilapia (Oreochromis niloticus) vitellogenins: development of homologous and heterologous ELISAs and analysis of vitellogenin pathway through the ovarian follicle

Journal of Experimental Zoology. Part A, Comparative Experimental Biology
Pap NdiayeJesús Núñez

Abstract

Vitellogenin (VTG) of Oreochromis niloticus was again purified, due to the conflicting results found in the literature. Three purification processes have been used: electrophoresis and electro-elution, double chromatography (gel filtration and ion-exchange chromatography) and single ion-exchange chromatography. Using SDS-PAGE we confirmed in all cases the presence of two polypeptidic forms of plasma VTG of 130 kDa (VTG1) and 170 kDa (VTG2). We raised polyclonal antibodies against each VTG form and we demonstrated the complete cross-reactivity of each antibody with both forms of VTG by Enzyme Immuno-Assay (EIA) and Western blots. The homologous ELISAs developed exhibited a detection limit of 6 ng x ml(-1), equivalent to 60 ng x ml(-1) of plasma VTG and allowed us to quantify the total plasma VTG of O. niloticus with high specificity and sensitivity. Using photonic and electron immunomicroscopy, we followed the pathway of VTG into the ovarian follicle (OF) demonstrating that VTG enters the oocyte at stage 3 of OF development, at the same time as cortical alveoli and lipid globules appear. Heterologous ELISAs performed on other cichlid species allowed us to quantify plasma VTG in Oreochromis aureus and Sarotherodon melanotheron an...Continue Reading

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Citations

Aug 1, 2008·Fish Physiology and Biochemistry·Jesús Núñez, Fabrice Duponchelle
Mar 8, 2014·Micron : the International Research and Review Journal for Microscopy·Irani Quagio-GrassiottoAriene Cristina Dias Guimarães-Bassoli
Jul 6, 2010·General and Comparative Endocrinology·Ha Ryong KimSeung Min Oh
Apr 2, 2021·General and Comparative Endocrinology·Rodolfo AmthauerLuis Vargas-Chacoff

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