2001 Fiscal Year Final Research Report Summary
Molecular mechanism of the regulation of neuronal nitric oxide synthase activity and its role in the regulation of the autonomic nervous system
| Project/Area Number |
12670058
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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 |
Environmental physiology (including Physical medicine and Nutritional physiology)
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| Research Institution | Osaka University |
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Principal Investigator |
HASHIDA Akiko (橋田 明子) Osakal Uniyersity, Institute for Protein Research, Instructor, 蛋白質研究所, 助手 (30180813)
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| Co-Investigator(Kenkyū-buntansha) |
OKUMURA Nobuaki Osakal Uniyersity, Institute for Protein Research, Assistant Professor, 蛋白質研究所, 助教授 (20224173)
NAGAI Katsuya Osakal Uniyersity, Institute for Protein Research, Professor, 蛋白質研究所, 教授 (70029966)
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| Project Period (FY) |
2000 – 2001
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| Keywords | neuronal nitric oxide synthase / α1-syntrophin / SCN |
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| Research Abstract |
This study shows the following results.
1. Role of α1-syntrophin in the activity and localization of neuronal nitric oxide synthase (nNOS)
The expression plasmids of nNOS and α1-syntrophin were transfected into COS-7 cells and the cGMP level in those cells was measured. Results showed that α1-syntrophin decreased the nitric oxide synthase activity. Then the images of the cells expressing both proteins were observed with a confocal microscope. Neuronal NOS was colocalized with α1-syntrophin. in the mitochondria, the endoplasmic reticulum and the Golgi body.
2. Colocalization of α1-syntrophin with neurotransmitters in the hypothalamus
In the hypothalamus, α1-syntrophin was localized in several arginine-vasopressin positive neurons in the hypothalamic paraventricular nucleus and suprachiasmatic nucleus. The results indicate that the complex made of nNOS and α1-syntrophin might play a role in the arginine-vasopressin secretion.
3. Purification and identification of α1-syntrophin binding proteins
To investigate α1-syntrophin binding proteins, pulldown assay was performed using the beads bound to various domains of α1-syntrophin fused with GST. Several α1-syntrophin binding proteins were detected as a result of the analysis. Identification of these proteins is in progress.
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