Production and characterization of antibodies to neuroparsins A and B isolated from the corpora cardiaca of the locust

Abstract

Antisera were raised in rabbits against neuroparsins A and B which were purified to near homogeneity using electroelution from 7,5% polyacrylamide electrophoresis gels. They were characterized using immunohistochemical, enzyme-linked immunosorbent assay (ELISA), and protein-blotting methods. The antineuroparsin A and the antineuroparsin B sera have different titers and sensitivities (higher titer for antineuroparsin A, higher sensitivity for antineuroparsin B). They exhibit very good specificity. The immunohistochemical study of the entire central nervous system using either antineuroparsin A or antineuroparsin B sera shows that only the A1 type of the protocerebral median neurosecretory cells are immunostained. Moreover, among the numerous proteins of the median region of the brain and of the corpora cardiaca, each immune serum recognized only neuroparsin A or neuroparsin B. Displacement curves obtained for each immune serum by competition between either neuroparsin A or neuroparsin B demonstrate that the neuroparsin A is recognized as well as neuroparsin B, with both antisera supporting the concept that these two proteins are chemically related. The nonspecific binding of neuroparsins to an antisomatostatin immune serum used at 1 100 dilution indicates thay any cross-reactivities of invertebrate molecules obtained with very low dilutions of antisera to vertebrate molecules must be considered carefully before concluding any immunological relation between invertebrate and vertebrate products.