| Project/Area Number |
14370473
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Orthopaedic surgery
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| Research Institution | Kyoto Prefectural University of Medicine |
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Principal Investigator |
OKAJIMA Seiichiro Kyoto Prefectural University of Medicine, Graduate school of medical science, Lecturer, 医学研究科, 講師 (70305580)
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| Co-Investigator(Kenkyū-buntansha) |
YAOI Takeshi Kyoto Prefectural University of Medicine, Graduate school of medical science, Teaching assistant, 医学研究科, 助手 (40311914)
FUSHIKI Shinji Kyoto Prefectural University of Medicine, Graduate school of medical science, Professor, 医学研究科, 教授 (80150572)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥10,200,000 (Direct Cost: ¥10,200,000)
Fiscal Year 2003: ¥4,600,000 (Direct Cost: ¥4,600,000)
Fiscal Year 2002: ¥5,600,000 (Direct Cost: ¥5,600,000)
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| Keywords | Neuromuscular junction / Schwann cell / Single cell derived total cDNA / Lysophosphatidic acid / Sphingosine-1-phosphate / Degenerate PCR / G-protein coupled receptors / LPA Receptors |
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| Research Abstract |
1.Gene analysis of the terminal Schwann cells specific LPA receptor family
Single terminal Schwann cell (TSC) and myelinating Schwann cell (MSC) were isolated from the sciatic nerves of adult Wistar rats. Total RNA from each single cell was purified and total cDNAs from each type of single cells were synthesized and amplified. We have checked the quality of cDNA fractions with S100β as a common marker for Schwann cells. The cDNA fractions containing S100β were used for the following PCR.
We performed PCR using the amplified cDNAs as templates and using specific primers for LPA receptors, to identify members of LPA receptors specifically expressed in TSCs. PCR analysis revealed that 1pA3 and 1pB1 were specifically expressed in TSC.
Using in situ hybridization histochemistry, we examined the expression patterns of members of LPA receptors in soleus muscles of rats. Specific signals for 1pA3 and 1pB1 mRNAs were detected in TSCs.
2.Serching of novel subtype of LPA receptors expressed in terminal Schwann cell
We performed PCR amplification of TSC cDNA using degenerate oligonucleotide primers against amino acid sequences from transmembrane domain. We are now identifying the novel subtype of LPA receptors.
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