A MONOCLONAL-ANTIBODY TO A 56 KDA NEUROSECRETORY POLYPEPTIDE FROM THEPOSTERIOR LATERAL NEUROSECRETORY-CELLS OF THE HOUSEFLY, MUSCA-DOMESTICA

A protocol was developed for generating monoclonal antibodies (MABs) t o neurosecretory cells (NSCs) of the housefly, Musca domestica, using head extract as immunogen. A panel of MABs to housefly NSCs was produc ed using a 3-tier screening involving enzyme-linked immunosorbent assa y (ELISA), western blot, and immunocytochemistry. One clone designated 01C10 was shown to immunostain the posterior lateral NSCs of the adul t housefly brain. Immunostaining of neurohemal organs, corpora cardiac a (CC), was restricted to the late pharate adult stage and the first 6 h of adult eclosion. Loss of immunostained material from CC indicated its release into the hemolymph. Western blot studies revealed that 01 C10 MAB immunoreacted with a 56 kDa polypeptide in the brain, corpora cardiaca/corpora allata (CC/CA) complex of newly emerged adults and he molymph collected 1 h after adult eclosion. These results strongly sug gested the neurohormonal nature of the 01C10-immunoreactive polypeptid e. We speculated the 56 kDa polypeptide to be bursicon based on a HPLC separation of hemolymph proteins from newly emerged adults which show ed the co-elution of bursicon activity and the 56 kDa polypeptide in t he same fractions.