| Project/Area Number |
18570075
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Animal physiology/Animal behavior
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| Research Institution | Fukuoka University |
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Principal Investigator |
SHIMOHIGASHI Miki Fukuoka University, Faculty of Science, Senior Assistant Professor (60078590)
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| Co-Investigator(Kenkyū-buntansha) |
SHIMOHIGASHI Yasuyuki Kyushu University, Faculty of Science, Professor (00211293)
MATSUMOTO Akira Kyushu University, Center for Research and Advancement in Higher Education, Assistant Professor (40229539)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,830,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥330,000)
Fiscal Year 2007: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2006: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | Circadian Rhythm / Insect / Apia mellifera / Drosophila / Pace maker hormone / PDF / 概目リズム |
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| Research Abstract |
Pigment-dispersing factor (PDF) is a pacemaker hormone regulating the locomotor rhythm in insects. The aim of this study was to clarify the mechanism to exhert the circadian oscilation between gene expression and paracrine exocytocis of PDF at the axonal terminal. First of all, in order to clarify whether the expression of pdf gene exhibit the circadian rhythm, we cloned the cDNAs encoding Apis PDF precursor protein, and found that there are at least seven different pdf mRNAs afforded by an alternative splicing site and five alternative polyadenylation sites in its 5'UTR and 3'UTR, respectably. The quantitative real-time PCR revealed the expression levels of each alternative splicing mRNA variant (5'UTR insertion-type and 5'UTR deletion-type) exhibit the daily fluctuation with a peak at the end of the day and that the expression profiles of both types of pdf mRNAs are similar each other. The whole mount in situ hybridization revealed the cells in the optic lobe of the brain express the
… More
se mRNA variants. Further experiments by the serial semi-thin sectioning identified the precise location and number of the expressing cells. The cells expressing the insertion type of pdf mRNAs were found to be 12 in hemisphere, and those of deletion type were 13. Since there are fourteen cells expressing any types of pdf mRNAs, these results implied there are three types expressing cells of pdf mRNAs.
PAP is the other neuropeptide encoded in the cis region of pdf locus. In order to investigate the function of PAP, we established three kinds of transgenic flies by using P-element mediated transfomation. One expresses only PAP driven by the native promoter of PDF locus, another does only pdf, and the remaining does both. Then we selected transgenic lines carrying each transgene only in the second chromosome, and established the homozygous transgenic strain in the pdf[01] background. We recorded their locomotor activity rhythms. The full length transgene of PDF locus rescued the arrhythmic phenotype of pdf[01], while the remaining two didn't. This suggests that not only PDF but also PAP are necessary to establish a normal circadian locomotion. Less
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