Gene targeting approach to study functions on respiratory and circulatory regulation of novel H^+ Channel
| Project/Area Number |
11680779
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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 |
Neuroscience in general
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| Research Institution | Gunma University |
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Principal Investigator |
SHIMOKAWA Noriaki Gunma Univ School of Med, Assistant Prof, 医学部, 講師 (90235680)
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| Project Period (FY) |
1999 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2001: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2000: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1999: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | Differential display / Gene targeting / Ventral medullary surface / H^+-sensitivity / Hypercapnia / Rhombex-29 / 40 / MafG / CO_2 / H^+sensitivity / hypercapnia / Transmembrane Protein / H^+ sensitivity / Maf protein / proteolipid protein / Mafg / Naclear Transcription Factor / Ventral Medullary Surface / Differential Display / H^+ーSensing |
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| Research Abstract |
1. Characterization and gene targeting study of ASPT
[1] ASPT comprises 751 amino acids containing 12 membrane-spanning helices
[2] ASPT has H^+/Sugar transport motif, Proline-rich region, leucine zipper motif.
[3] ASPT immunoreactive neurons were found in the VMS and the number of cells was increased by hypercapnic stimulation.
[4] We got clones containing exons of ASPT gene from genomic library of ES cells.
[5] We constructed a vector for targeting of ASPT gene.
[6] We obtained ES cells that generated homologous recombination.
2. Detection of novel genes involved in H^+-sensitivity in the VMS
The ventral medullary surface (VMS) in known as the site of the central chemosensitive neurons. These neurons sense excess CO_2/H^+ dissolved in the cerebrospinal fluid that superfuses the VMS and induce hyperventilation. We hypothesized that genes specific for hyperventilation are expressed much more highly in VMS neurons than in extra-VMS neurons in other parts of the central nervous system (CNS). Applying the differential display technique to the brain of adult rats, we differentiated the mRNAs of the VMS neurons from those of cerebral cortex neurons.
[1] We cloned a novel four-transmembrane protein, Rhombex-29. Structural analysis and the phylogenic tree showed that rat Rhombex-29 is homologous to the major CNS myelin protein PLP/DM20-M6 family (REFERENCES 1).
[2] We cloned cDNA encoding nuclear transcription factor MafG from rat VMS (REFERENCES 2).
[3] We got clones encoded a putative transmembrane protein, rhombencephalic expression protein-40 kDa (Rhombex-40) (REFERENCES 3).
[4] We cloned MafG-2, a novel splice variant of MafG, from rat brain by RT-PCR (REFERENCES 4).
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Report
(4 results)
Research Products
(14 results)
