| Project/Area Number |
16591912
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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 |
Conservative dentistry
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| Research Institution | Okayama University |
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Principal Investigator |
TASHIRO Yoko Okayama university hospital, associate researcher, 医学部・歯学部附属病院, 助手 (20362981)
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| Co-Investigator(Kenkyū-buntansha) |
IZAWA Shunji Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, associate researcher, 大学院・医歯薬学総合研究科, 助手 (20273998)
TORII Yasuhiro Okayama university hospital, professor, 医学部・歯学部附属病院, 教授 (10188831)
TAKABATAKE Yasumitsu Okayama university hospital, associate researcher, 医学部・歯学部附属病院, 助手 (10362980)
YOSHIYAMA Masahiro Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, professor, 大学院・医歯薬学総合研究科, 教授 (10201071)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 2005: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2004: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | nucleus of the solitary tract / area postrema / dopamine receptor / RT-PCR / immunohistochemistry / in situ hybridization / rat / ドーパミン受容体mRNA / 免疫染色 |
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| Research Abstract |
The present study elucidated the localization of the dopamine D_2 and D_5 receptors and mRNAs in the rat area postrema and the nucleus of the solitary tract using RT-PCR, immunohistochemistry and in situ hybridization. Dopamine is an important central nervous system neurotransmitter that exerts many physiological activities, and animal studies suggest that the dopaminergic system plays a role in central pain modulation. In view of this, the area postrema and the nucleus of the solitary tract within the medulla oblongata may play a role in central pain modulation, suggesting that it may be involved in central pain control. RT-PCR Male Sprague-Dawley albino rats were anesthetized with halothane and then decapitated. The area postrema and the nucleus of thee solitary tract was surgically resected with sterilized incision. Immediately after resection, it was put into a sterilized tube containing ISOGEN to extract total RNA. For reverse transcription, RNA was processed and PCR was performed. The reaction was amplified and PCR amplicons of the samples were visualized on ethidium bromide containing agarose gel. Immunohistochemistry For sectioning, the sample was cut at 10 μm thickness with a cryostat microtome. Sections were probed with rabbit polyclonal antibody against rat dopamine receptor subtypes D2 and 5. The probed sections were detected using avidin-biotin complex and diaminobenzidine. in situ hybridization Single stranded digoxigenin labeled antisense or sense RNA probes for D2 and D5 receptors were generated. For in situ hybridization, 10 μm thickness sections were used. Hybridization was performed overnight at 55℃ in a humidified chamber. Briefly, the slides were incubated with anti-digoxigenin-AP, Fab fragment and then stained with a solution of NBT and BCIP. Subsequently, developed sections were rinsed with distilled water, counterstained with 2% methyl green and coverslipped in Pristine Mount. Slides were examined on a Olympus BX40 microscope.
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