Molecular neuro-physiological study of insect diapause mechanism

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PCR was carried out using cDNA prepared from brain of the silkworm, Bombyx mori with primers by which opsin encoding genes of the compound eyes can be amplified. We got an amplification of a band corresponding about 200 bases, and then subcloned a fragment of 210 bases DNA (70 amino acids) encoding a opsin-like gene. On the base of this DNA sequence, we conducted 3'-5'-Race method to determine all length of ORF region. The cloned DNA sequence had about 1600 bases and the ORF coded 377 amino acids. It was found that the DNA sequence has a high homology with ManipI reported as one of the photoreceptive pigments in the Manduca sexta compound eyes. We named this opsin like protein in the silkworm brain Boceropisn (bombyx cerebral opsin). One peptide corresponding to the carboxyterminal 25 amino acids of Boceropsin was synthesized. And immunocytochemistry revealed that some pairs of cells of the silkworm brain showed potent immunoreactive to the rabbit antisera of the peptide. These immunoreactive cells is suggested to be a pupative photoreceptor involved in the photoperiodic response in the silkworm.