Partial identification, synthesis and immunolocalization of locustamyoinhibin, the third myoinhibiting neuropeptide isolated from Locusta migratoria

Abstract

A blocked neuropeptide that suppresses the motility of the cockroach hindgut has been isolated from an extract of 9000 brain-corpora cardiaca-corpora allata-suboesophageal ganglion complexes of Locusta migratoria. Biological activity was monitored during HPLC purification by observing the myoinhibiting activity of column fractions on the isolated hindgut of Leucophaea maderae. Due to the low amount of material left after deblocking, this myoinhibiting peptide--designated as locustamyoinhibin or Lom-MIH--could only be partially sequenced: pGlu-X-Tyr-X'-Lys-Gln-Ser-Ala-Phe-Asn-Ala-Val-Ser-NH2. Nevertheless, the carboxy-terminal nonamer sequence (Lom-MIH5-13) was synthesized and also displayed myoinhibiting activity, indicating that the biologically active core lies in the carboxy-terminal sequence. Lom-MIH shows no sequence similarities with other peptides from vertebrate or invertebrate sources and is the third myoinhibiting peptide identified in Locusta migratoria. A polyclonal antiserum was raised against Lom-MIH5-13 and used to investigate the distribution of immunoreactive peptide in the central nervous system and its associated neurohaemal structures. Two groups of neurons with somata in the optic lobes show locustamyoinhibin (Lom-MIH)-like immunoreactivity. These groups have somata at the dorsal and ventral edge of the lamina ganglionaris. The neurons have dense ramifications in the lamina, with processes extending into the first optic chiasma and into the accessory medulla. Four cell bodies were detected in the protocerebrum, and two cells were found at the externo-lateral edge of the tritocerebrum. No immunoreactive perikarya could be observed in the suboesophageal ganglion nor in the ganglia of the ventral nerve cord. Neither the corpora cardiaca nor the neurohaemal organs of the ventral nerve cord showed immunolabelling. Therefore, our findings provide anatomical evidence for a central neurotransmitter role of Lom-MIH.