2005 Fiscal Year Final Research Report Summary
A positional cloning study of the responsible gene for Pma, a spontaneous mutant mouse.
| Project/Area Number |
15500227
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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 |
Nerve anatomy/Neuropathology
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| Research Institution | University of Tsukuba |
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Principal Investigator |
MASU Kazuko University of Tsukuba, Grad.Sch.Comprehensive Human Sci., Assistant Prof., 大学院・人間総合科学研究科, 講師 (50344883)
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| Co-Investigator(Kenkyū-buntansha) |
MASU Masayuki University of Tsukuba, Grad.Sch.Comprehensive Human Sci., Professor, 大学院・人間総合科学研究科, 教授 (20243032)
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| Project Period (FY) |
2003 – 2005
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| Keywords | Motor Nerves / Development / Genes / Genome / mutant mouse |
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| Research Abstract |
In this study, I aimed to elucidate the molecular mechanism that regulates the axon guidance by identifying the responsible gene for a spontaneous mutant mouse, which was discovered at the Central Institute of Experimental Animals, which has genetic defect in a motor nerve projection.
I mapped the responsive gene within a 〜2 Mb region of a certain chromosome in a previous linkage analysis study. For more precise mapping, I generated 257 new microsatellite markers. After examination of these markers to distinguish the mutant allele from normal one's, I further narrowed the critical region by a linkage analysis using these markers and more samples. Database search revealed that there are about 20 predicted transcripts within this region, and that this region does not include genes such as EphA4,GDNF, and c-ret.
I then examined the expression of these genes in the normal mouse embryos by in situ hybridisation, and analysed the structure of the genes that are expressed in the mouse embryos.
So far, I determined and compared the nucleotide sequences of the 222 predicted exons and surrounding regions from the mutant mouse and 2 normal mouse strains (C57BL/6 and 129/Sv). I found 70 one-base-substitutions and 3 small deletions in exons,179 one-base-substitutions and 16 small deletions or insertions in intronic sequences. However, no typical mutations that lead to truncation or deficit of a protein were identified, although there were some mutations inducing amino acid substitutions.
Next, I compared expression of 16 predicted genes in the candidate region in mutant and normal mice using both in situ hybridization and RT-PCR. Two genes appeared to be reduced in expression levels. The research is under way to analyze these two genes as strong candidates for the responsible gene of the mutant phenotype.
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Research Products
(17 results)
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[Journal Article] Genetic marking of hematopoietic stem and endothelial cells : Identification of the Tmtsp genes encoding a novel cell-surface protein with the thrombospondin-1 domain.
Author(s)
Takayanagi S, Hiroyama T, Yamazaki S, Nakajima T, Morita Y, Usui J, Eto K, Motohashi T, Shiomi K, Keino-Masu K, Masu M, Oike Y, Mori S, Yoshida N, Iwama A, Nakauchi H.
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Journal Title
Description
「研究成果報告書概要(欧文)」より
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