Journal of Neuroscience, Vol 6, 1738-1747, Copyright © 1986 by Society for Neuroscience
Identification of the cerebral neurosecretory cells that contain eclosion hormone in the moth Manduca sexta
PF Copenhaver and JW Truman
Eclosion hormone (EH) is an insect neuropeptide that is released at the end
of metamorphosis from the CNS and triggers the stereotyped motor program of
adult emergence. Using three distinct experimental approaches, we have
identified a discrete set of neurosecretory cells in the brain of the moth
Manduca sexta that contains and releases EH. By isolating the
neurosecretory somata and testing them with a sensitive behavioral
bioassay, we identified a cluster of ipsilaterally projecting cells (Group
Ia) that contain EH. Intracellular stimulation of individual cells within
this group induced the release of bioactive EH into the hemolymph
surrounding the neurohemal organs of the brain, whereas stimulation of
cells in the other cerebral neurosecretory clusters did not. We also
developed a polyclonal antiserum against purified EH that precipitated all
bioactive material from samples containing the peptide. This antiserum
selectively stained 5 of the Group Ia cells on either side of the brain, as
well as their central and terminal processes. Preincubation of the serum
with EH dramatically reduced its ability to bind the peptide subsequently.
The combined application of these physiological and immunological
techniques has led to the unequivocal identification of the EH neurons in
the moth brain.
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