title: A circadian neuropeptide PDF in the honeybee, Apis mellifera: cDNA cloning and expression of mRNA.
abstract: Pigment-dispersing factor (PDF) is a pacemaker hormone regulating the locomotor rhythm in insects. In the present study, we cloned the cDNAs encoding the Apis PDF precursor protein, and found that there are at least seven different pdf mRNAs yielded by an alternative splicing site and five alternative polyadenylation sites in the 5'UTR and 3'UTR regions. The amino acid sequence of Apis PDF peptide has a characteristic novel amino acid residue, aspargine (Asn), at position 17. Quantitative real-time PCR of total and 5'UTR insertion-type pdf mRNAs revealed, for the first time, that the expression levels change in a circadian manner with a distinct trough at the beginning of night in LD conditions, and at the subjective night under DD conditions. In contrast, the expression level of 5'UTR deletion-type pdf mRNAs was about half of that of the insertion type, and the expression profile failed to show a circadian rhythm. As the expression profile of the total pdf mRNA exhibited a circadian rhythm, transcription regulated at the promoter region was supposed to be controlled by some of the clock components. Whole mount in situ hybridization revealed that 14 lateral neurons at the frontal margin of the optic lobe express these mRNA isoforms. PDF expressing cells examined with a newly produced antibody raised against Apis PDF were also found to have a dense supply of axon terminals in the optic lobes and the central brain.
authors: Division of Biology, Department of Earth System of Science, Faculty of Science, Fukuoka University, Fukuoka 814-0180, Japan.
authors: Miho
authors: M
authors: Sumiyoshi
authors: Seiji
authors: S
authors: Sato
authors: Yukimasa
authors: Y
authors: Takeda
authors: Kazunori
authors: K
authors: Sumida
authors: Keita
authors: K
authors: Koga
authors: Tsunao
authors: T
authors: Itoh
authors: Hiroyuki
authors: H
authors: Nakagawa
authors: Yasuyuki
authors: Y
authors: Shimohigashi
authors: Miki
authors: M
authors: Shimohigashi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001516:Bees / Q000235:genetics / Q000378:metabolism*; D002874:Chromosome Mapping; D059006:Chromosomes, Insect / Q000235:genetics; D002940:Circadian Rhythm / Q000502:physiology; D018076:DNA, Complementary / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000502:physiology*; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000235:genetics / Q000378:metabolism; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*
chemical_list: D018076:DNA, Complementary; D019476:Insect Proteins; D009479:Neuropeptides; D010455:Peptides; D012333:RNA, Messenger; C027714:melanophore-dispersing hormone