Molecular cloning and circadian expression profile of insect neuropeptide PDF in black blowfly, Phormia regina

Ayami Matsushima, Satoru Yokotani, Xiaohui Lui, Kazunori Sumida, Takeshi Honda, Seiji Sato, Atsushi Kaneki, Yukimasa Takeda, Yoshiro Chuman, Mamiko Ozaki, Daisuke Asai, Takeru Nose, Hitoshi Onoue, Yushi Ito, Yoshiya Tominaga, Yasuyuki Shimohigashi, Miki Shimohigashi

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Pigment-dispersing factor PDF is an 18-amino acid insect neuropeptide that mediates a circadian rhythmicity in locomotor activity. PDF is coded in a precursor protein together with another neuropeptide named PDF-associated peptide, PAP. PDF is highly conserved among insects, whereas the homology of PAPs is very low with considerably varied amino acid sequences. Since such dissimilarity has suggested that the function of PAP peptide is not associated with that of PDF, we have attempted to analyze the sequences of PDF precursor proteins among a series of species of insects and hypothesized that PDF precursors are classified into at least three different classes: Drosophila-Musca, Meimuna-Romalea, and Gryllus. In order to exemplify this hypothesis, we here describe the molecular cloning of the pdf-gene of the black blowfly Phormia regina and an in silico screening for the pdf-gene in the genome databank of the mosquito Anopheles gambie, both species belonging to the Diptera. It was found that deduced amino acid sequences of PDF peptides are almost completely conserved among all Dipterans and also the amino acid sequences of PAPs are considerably highly preserved (55-82 similarity) among the species of Diptera. The results confirmed the validity of grouping the PDF precursor proteins. In situ hybridization was carried out in fly brains to identify the precise locations of pdf-expressing cells and to examine any daily cycling of pdf mRNA. Intense signals for pdf mRNA were identified in the medulla, but not in the pars lateralis where PDF neurons were strongly immunostained by the antibody raised against PDF peptide. Hybridization was also performed for the brain samples at two hour intervals throughout the day. Although very intense hybridization signals were observed at ZT8 even in some neurites, no prominent rhythmicity of pdf mRNA expression was observed.

Original languageEnglish
Pages (from-to)419-430
Number of pages12
JournalLetters in Peptide Science
Volume10
Issue number5-6
DOIs
Publication statusPublished - Nov 1 2003

Fingerprint

Cloning
Molecular Cloning
Neuropeptides
Peptides
Protein Precursors
Insects
Amino acids
Diptera
Amino Acids
Amino Acid Sequence
Genes
Periodicity
Proteins
Messenger RNA
Brain
Anopheles
Neurites
Locomotion
Culicidae
Reproducibility of Results

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Molecular cloning and circadian expression profile of insect neuropeptide PDF in black blowfly, Phormia regina. / Matsushima, Ayami; Yokotani, Satoru; Lui, Xiaohui; Sumida, Kazunori; Honda, Takeshi; Sato, Seiji; Kaneki, Atsushi; Takeda, Yukimasa; Chuman, Yoshiro; Ozaki, Mamiko; Asai, Daisuke; Nose, Takeru; Onoue, Hitoshi; Ito, Yushi; Tominaga, Yoshiya; Shimohigashi, Yasuyuki; Shimohigashi, Miki.

In: Letters in Peptide Science, Vol. 10, No. 5-6, 01.11.2003, p. 419-430.

Research output: Contribution to journalArticle

Matsushima, A, Yokotani, S, Lui, X, Sumida, K, Honda, T, Sato, S, Kaneki, A, Takeda, Y, Chuman, Y, Ozaki, M, Asai, D, Nose, T, Onoue, H, Ito, Y, Tominaga, Y, Shimohigashi, Y & Shimohigashi, M 2003, 'Molecular cloning and circadian expression profile of insect neuropeptide PDF in black blowfly, Phormia regina', Letters in Peptide Science, vol. 10, no. 5-6, pp. 419-430. https://doi.org/10.1007/BF02442573
Matsushima, Ayami ; Yokotani, Satoru ; Lui, Xiaohui ; Sumida, Kazunori ; Honda, Takeshi ; Sato, Seiji ; Kaneki, Atsushi ; Takeda, Yukimasa ; Chuman, Yoshiro ; Ozaki, Mamiko ; Asai, Daisuke ; Nose, Takeru ; Onoue, Hitoshi ; Ito, Yushi ; Tominaga, Yoshiya ; Shimohigashi, Yasuyuki ; Shimohigashi, Miki. / Molecular cloning and circadian expression profile of insect neuropeptide PDF in black blowfly, Phormia regina. In: Letters in Peptide Science. 2003 ; Vol. 10, No. 5-6. pp. 419-430.
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abstract = "Pigment-dispersing factor PDF is an 18-amino acid insect neuropeptide that mediates a circadian rhythmicity in locomotor activity. PDF is coded in a precursor protein together with another neuropeptide named PDF-associated peptide, PAP. PDF is highly conserved among insects, whereas the homology of PAPs is very low with considerably varied amino acid sequences. Since such dissimilarity has suggested that the function of PAP peptide is not associated with that of PDF, we have attempted to analyze the sequences of PDF precursor proteins among a series of species of insects and hypothesized that PDF precursors are classified into at least three different classes: Drosophila-Musca, Meimuna-Romalea, and Gryllus. In order to exemplify this hypothesis, we here describe the molecular cloning of the pdf-gene of the black blowfly Phormia regina and an in silico screening for the pdf-gene in the genome databank of the mosquito Anopheles gambie, both species belonging to the Diptera. It was found that deduced amino acid sequences of PDF peptides are almost completely conserved among all Dipterans and also the amino acid sequences of PAPs are considerably highly preserved (55-82 similarity) among the species of Diptera. The results confirmed the validity of grouping the PDF precursor proteins. In situ hybridization was carried out in fly brains to identify the precise locations of pdf-expressing cells and to examine any daily cycling of pdf mRNA. Intense signals for pdf mRNA were identified in the medulla, but not in the pars lateralis where PDF neurons were strongly immunostained by the antibody raised against PDF peptide. Hybridization was also performed for the brain samples at two hour intervals throughout the day. Although very intense hybridization signals were observed at ZT8 even in some neurites, no prominent rhythmicity of pdf mRNA expression was observed.",
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AU - Honda, Takeshi

AU - Sato, Seiji

AU - Kaneki, Atsushi

AU - Takeda, Yukimasa

AU - Chuman, Yoshiro

AU - Ozaki, Mamiko

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AU - Nose, Takeru

AU - Onoue, Hitoshi

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