Study summary · research use only
Production and immunohistochemical application of monoclonal antibodies against delta sleep-inducing peptide
Plain-language summary
Paraphrased from the published abstract below — not a verdict on whether anything works.
In this study, the authors produced monoclonal antibodies by immunizing rats with delta sleep-inducing peptide (DSIP) and fusing spleen cells with SP2/0 myeloma cells. Screening by solid-phase immunoassay identified six stable clones reacting with DSIP, all recognizing epitopes in the carboxy-terminal region DSIP5-9. Applied to rat median eminence sections, the antibodies strongly labeled fibres and terminal-like structures, mainly in lateral areas, and their pattern overlapped almost completely with luteinizing hormone-releasing hormone immunoreactivity, though the authors argue the two systems are independent. The antibodies did not label median eminence in mouse, hamster, or gerbil, which the authors interpret as species differences in DSIP5-9-related epitopes.
Abstract
Monoclonal antibodies were produced following immunization of rats with delta sleep-including peptide (DSIP). The spleen cells of the rats were fused with the myeloma cell line SP2/0. The supernatants of hybridomas were screened on a solid-phase immunoassay using dot-immunobinding of DSIP and some DSIP fragments. The supernatants of six stable producer clones were found to react with DSIP. From this procedure it was also deduced that all these monoclonal antibodies recognized epitope(s) of the penta carboxy-terminal region of DSIP (DSIP5-9). Application of these monoclonal antibodies to rat median eminence sections gave a strong immunolabelling of a large population of fibres and terminal-like structures, mainly localized through the lateral areas. Elution-restaining experiments using a monoclonal antibody to DSIP and a polyclonal antiserum to luteinizing hormone-releasing hormone (LHRH) showed that the patterns of immunoreactivity respectively visualized overlap almost completely. Although numerous LHRH-immunoreactive neuronal elements were also easily demonstrated in the median eminence of the mouse, the hamster and the gerbil species, incubation of sections with monoclonal antibodies to DSIP failed to give any immunoreaction. Taken together these data argue for the independence of the DSIP/LHRH immunolabelling systems. Furthermore, it was demonstrated that DSIP5-9-related epitopes detected in the rat median eminence have no counterpart in the three other rodent species investigated. These species differences may reflect the fact that the carboxy-terminal sequence of the nonapeptide DSIP originally discovered in the rabbit is not conserved in all rodent species.
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