2007 Fiscal Year Final Research Report Summary
Molecular characterization of mutant protein kinase C gamma found in spinocerebellar ataxia type 14 and its application to screening of therapeutics
| Project/Area Number |
18500296
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Neurochemistry/Neuropharmacology
|
|---|
| Research Institution | Hiroshima University |
|---|
Principal Investigator |
SAKAI Norio Hiroshima University, Grad Sch. of Biomed Sci, Professor (70263407)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
SAKI Takahiro Hiroshima univ, Graduate School of Biomedical Sciences, Assistant Professor (50335650)
|
|---|
| Project Period (FY) |
2006 – 2007
|
| Keywords | spinocerebellar ataxia / rotein kinase / ubioutin proteasome system / aggregation / ER stress / apoptosis |
|---|
| Research Abstract |
1. Several causal missense mutations in protein kinase CY (γPKC) gene have been found in spinocerebellar ataxia type 14 (SCA14), an autosomal dominant neurodegenerative disease. We previously demonstrated that mutant γPKC found in SCA14 is susceptible to aggregation and induces apoptosis in cultured cell lines In the present study, we examined whether mutant γPKC formed aggregation and how mutant γPKC affect the morphology and survival of cerebellar Purkinje cells (PCs), which are degenerated in SCA14 patients. Wild type (WT) and mutant γPKG-GFP were expressed in PCs of mouse cerebellar primary culture by using adenoviral vectors Aggregates of mutant γPKC-GFP were also formed in PCs and were indispensable for PC apoptosis. However, the long-term time-lapse observation revealed that Its have a protective potential to eliminate the toxic aggregates of mutant γPKG-GFP and this potential appeared prominently after the withdrawal of mutant γPKG-GFP expression. Mutant γPKG-GFP disturbed the
… More
development of PC dendrites and reduced synapse formation, regardless of presence or absence of its aggregate. FRAP (Fluorescence recovery after photobleaching) analysis revealed the reduced mobility of mutant γPKG-GFP, which may cause the attenuated translocation of mutant γPKG-GFP upon the stimulation of high KC1 These results indicate that mutant γPKC not only forms toxic aggregates but also affects the dendritic development and the synaptic formation in PCs, probably due to the reduced mobility and the insufficient translocation of mutant γPKC. These characters of mutant γPKC in PCs would be involved in the atrophy of cerebellar cortex and subsequently caused cerebellar dysfunction in SCA14 patients.
2. Several causal missense mutations in protein kinase Cγ (γPKC) gene have been found in spinocerebellar ataxia type 14 (SCA14), an autosomal dominant neurodegenerative disease. We previously demonstrated that mutant γPKC found in SCA14 is susceptible to two types of aggregation, cytoplasmic dot-like and perinuclear massive aggregation, and causes cell death in Chinese hamster ovary (CHO) cells. Long-term time-lapse imaging revealed that firstly-accumulated dot-Like aggregation of mutant γPKC-GFP gradually formed perinuclear massive aggregations, followed by cell death. However, it remains unclear how aggregate formation of mutant γPKC causes cell death In the present study, we examined whether these mutant aggregations affect the ubiquitin-proteasome system (UPS) and endoplasmic reticular (ER) strum. 'No mutant γPKG-GFPs (S119P and G128D) were strongly ubiquitinated, and dot-like aggregations of these mutants were ubiquitin-positive and colocalized with proteasome 20S. Furthermore, proteasome activity in cells with aggregates, especially massive ones, was significantly decreased. Aggregate formation of mutant γPKG-GFP induced phosphorylation of PERK (PKR-like ER kinase) and nuclear expression of CHOP (C/EBP homologous protein), hallmarks of ER stress and subsequently activated caspase-3. These results indicate that aggregate formation of mutant γPKC found in SCA14 impairs UPS and induces ER stress, leading to apoptotic cell death. Less
|
Research Products
(38 results)
-
-
[Journal Article] N-acetyl-l-aspartate activates hippocampal CA3 neurons in rodent slice preparations2008
Author(s)
Hanaya, R., Kiura, Y., Kurisu, K., Sakai, N., Serikawa, T., Sasa, M
-
Journal Title
Brain Res. Bull 75
Pages: 663-667
Description
「研究成果報告書概要(欧文)」より
-
-
-
-
-
-
[Journal Article] Fragmentation of protein kinase N (PKN)in the hydrocephalic rat brain Acta Histochem2007
Author(s)
Okii, N., Amano, T., Seki T., Matsubayashi, H., Mukai, H., Oma, Y., Kurisu, K., Sakai, N
-
Journal Title
Cytochem 40
Pages: 113-121
Description
「研究成果報告書概要(欧文)」より
-
[Journal Article] BDNF, NT-3, and NGF released from transplanted neural progenitor cells promote corticospinal axon growth in organotypic cocultures2007
Author(s)
Kamei, N., Tanaka, N.,Oishi, Y., Hamasaki, T., Nakanishi, K., Sakai, N., Ochi, M
-
Journal Title
Spine 32
Pages: 1272-1278
Description
「研究成果報告書概要(欧文)」より
-
[Journal Article] Bone marrow stromal cells promoting corticospinal axon growth through the release of humoral factors in organotypic cocultures in neonatal rats2007
Author(s)
Kamei, N, Tanaka, N., Oishi Y., Ishikawa, M., Hamasaki, T., Nishida, K., Nakanishi, K., Sakai, N., Ochi, M
-
Journal Title
J. Neurosurg Spine 20
Pages: 412-419
Description
「研究成果報告書概要(欧文)」より
-
[Journal Article] Magnetically labeled neural progenitor cells, which are localized by magnetic force, promote axon growth in organotypic cocultures2007
Author(s)
Hamasaki, T., Tanaka N., Kamei N., Ishida, O., Yanada, S., Nakanishi, K., Nishida, K., Oishi Y., Kawamata, S., Sakai, N., Ochi, M
-
Journal Title
Spine 32
Pages: 2300-2305
Description
「研究成果報告書概要(欧文)」より
-
[Journal Article] Aggregate formation of mutant protein kinase C gamma found in spinocerebellar ataxia type 14 impairs ubiquitin proteasome system and induces endoplasmic reticulum stress2007
Author(s)
Seki, T., Takahashi H., Adachi, N., Abe, N., Shimahara, T., Saito, N., Sakai, N
-
Journal Title
Eur. J. Neurosci 26
Pages: 3126-3140
Description
「研究成果報告書概要(欧文)」より
-
-
-
-
-
[Journal Article] dentification of a new family of spnocerebellar ataxia type 14 (SCA14) in the Japanese SCA population by the screening of PRKCG exon 42006
Author(s)
Hiramoto, K., Kawakami, H., Inoue, K., Seki, T, Maruyama H., Morino, H., Matsumoto M. Kurisu,K., Sakai, N
-
Journal Title
Movemeni Disorders 21
Pages: 1355-1360
Description
「研究成果報告書概要(欧文)」より
-
[Journal Article] Embryonic stem cell-derived neuror models of Parkinson's disease exhibit delayed neuronal death2006
Author(s)
Yamashita, H., Nakamura, T., Takahashi T., Nagano Y., Hiji, M., Hirabayashi T., Amano, T., Yagi, T., Sakai, N., Kohriyama, T., Matsumoto,M
-
Journal Title
J. Neurochem 98
Pages: 45-56
Description
「研究成果報告書概要(欧文)」より
-
[Journal Article] Microglial α7 nicotinic acetylcholine receptors drive a phospholipase C/IP3 pathway and modulate the cell activation towards a neuroprotective role2006
Author(s)
Suzuki, T.Hide, I., Matsubara,A.Hama, C., Harada, K.Miyano, K., Andra, M., Matsubayashi, H., Sakai,N., Kohsaka,S., Inoue,K., Nakata,Y
-
Journal Title
J. Neurosci Res 83
Pages: 1461-1470
Description
「研究成果報告書概要(欧文)」より
-
[Journal Article] R659S mutation of gamma PKC is susceptible to cell death : Implication of this mutation/polymorphism in the pathogenesis of retinitis pigmentosa2006
Author(s)
Mochizuki, H., Seki, T., Adachi, N., Saito, N., Mishima, H.K., Sakai, N
-
Journal Title
Neurochem. Int 49
Pages: 669-675
Description
「研究成果報告書概要(欧文)」より
-
[Journal Article] Fused protein of δPKC activation loop and PDK1-interacting fragment (δAL-PIF) functions as a pseudosubstrate and an inhibitory molecule for PDK1 when expressed in cells2006
Author(s)
Seki, T., Irie, N., Nakamura, K., Sakaue, H., Ogawa,W., Kasuga, M., Yamamoto, H., Ohmori, S., Saito, N., Sakai, N
-
Journal Title
Genes to Cells 11
Pages: 1051-1070
Description
「研究成果報告書概要(欧文)」より
-
-
-
-
-
-
-
[Presentation] Aggregate fromation and cytotoxic effect of mutant protein kinase Cy found in spinocerebellar ataxia 14 was prevented by disaccharide trehalose2007
Author(s)
Takahiro, Seki, Takayuki, Shimahara, Naoko, Adachi, Naoaki, Saito, Norio, Sakai
Organizer
the 37th annual meeting of the Society for Neuroscience
Place of Presentation
San Diego, USA
Year and Date
2007-11-05
Description
「研究成果報告書概要(欧文)」より
-
[Presentation] Effects of mutant yPKC found in SCA14 on the nature of primary-cultured Purkinje cells2007
Author(s)
Norio, Sakai, Takahiro, Seki, Takayuki, Shimaharai, Naoko, Adachi, Naoaki, Saito
Organizer
the 37th annual meeting of the Society for Neuroscience
Place of Presentation
San Diego, USA
Year and Date
2007-11-05
Description
「研究成果報告書概要(欧文)」より
-
-
-
-
-
-
-
-
-
