Mutant γPKC found in spinocerebellar ataxia type 14 induces aggregate-independent maldevelopment of dendrites in primary cultured Purkinje cells

Seki, Takahiro; Shimahara, Takayuki; Yamamoto, Kazuhiro; Abe, Nana; Amano, Taku; Adachi, Naoko; Takahashi, Hideyuki; Kashiwagi, Kaori; Saito, Naoaki; Sakai, Norio

doi:10.1016/j.nbd.2008.10.013

Cited by 58 publications

(94 citation statements)

References 63 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We constructed two types of adenoviral vectors to express FLAG-SERT under the regulation of tetracycline (32,33). The first vector expresses the tetracycline transactivator (tTA) under the control of the cytomegalovirus (CMV) promoter (Ad-CMV-tTA).…”

Section: Generation Of Plasmids and Adenoviral Vectorsmentioning

confidence: 99%

“…The experimental procedures are summarized in our previous report (32,33 3 H]5-HT was stopped by washing three times with cold KRH buffer containing 10 M fluvoxamine, and the cells were then lysed with RIPA buffer (10 mM TrisHCl, 1% NP-40, 0.1% SDS, 0.1% sodium deoxycholate, 150 mM NaCl and 1 mM EDTA pH 7.4). The cell extracts were diluted in a scintillation cocktail (Clear-Sol II; Nakalai Tesque, Kyoto), and their radioactivity was measured with a liquid scintillation counter (LSC-5100; Aloka, Mitaka).…”

Section: Cell Culture and Transfectionmentioning

confidence: 99%

“…Immunocytochemistry was carried out for FLAG-SERT-expressing HEK293 cells as previously described (18,32). In brief, the cells were washed in 1 ml PBS and fixed with 4% paraformaldehyde and 0.2% picric acid in 0.1 M phosphate-buffered saline (PBS, pH 7.4) for 30 min at RT, washed twice with 1 ml PBS, and then permeabilized with 500 l PBS supplemented with 0.3% Triton X-100 and 5% normal goat serum (NGS) for 15 min.…”

Section: Immunocytochemistry and Analysis Of Plasma Membrane Expressimentioning

confidence: 99%

See 2 more Smart Citations

Effects of the Chemical Chaperone 4-Phenylbutylate on the Function of the Serotonin Transporter (SERT) Expressed in COS-7 Cells

Fujiwara¹,

Yamamoto²,

Miyagi³

et al. 2013

J Pharmacol Sci

Self Cite

View full text Add to dashboard Cite

Abstract. The serotonin transporter (SERT) is involved in various psychiatric disorders, including depression and autism. Recently, chemical chaperones have been focused as potential therapeutic drugs that can improve endoplasmic reticulum (ER) stress-related pathology. In this study, we used SERT-transfected COS-7 cells to investigate whether 4-phenylbutylate (4-PBA), a chemical chaperone, affects the membrane trafficking and uptake activity of SERT. Treatment with 4-PBA for 24 h dose-dependently increased the uptake activity of SERT. In accordance with increased SERT activity, the expression of maturely glycosylated SERT was increased, while the expression of immaturely glycosylated SERT was decreased. This finding suggests that 4-PBA increased the functional SERT with mature glycosylation via accelerating its folding and trafficking. 4-PBA also increased the activity of the C-terminus-deleted mutant SERT (SERTCT), which was stacked in the ER, and decreased SERTCT-induced ER stress, further supporting the idea that 4-PBA acts as a chemical chaperone for SERT. Imaging studies showed that fluorescencelabeled SERT was gradually and significantly translocated to the plasma membrane by 4-PBA. These results suggest that 4-PBA and related drugs can potentially affect serotonergic neural transmission by functioning as chaperones, thereby providing a novel therapeutic approach for SERT-related diseases.[Supplementary materials: available only at http://dx

show abstract

Section: Generation Of Plasmids and Adenoviral Vectorsmentioning

confidence: 99%

Section: Cell Culture and Transfectionmentioning

confidence: 99%

Section: Immunocytochemistry and Analysis Of Plasma Membrane Expressimentioning

confidence: 99%

See 1 more Smart Citation

Effects of the Chemical Chaperone 4-Phenylbutylate on the Function of the Serotonin Transporter (SERT) Expressed in COS-7 Cells

Fujiwara¹,

Yamamoto²,

Miyagi³

et al. 2013

J Pharmacol Sci

Self Cite

View full text Add to dashboard Cite

show abstract

“…We have previously demonstrated that mutant versions of γPKC form aggregates in cultured cells (19) and mouse primary cultured PCs (20), leading to apoptotic cell death (21). Furthermore, mutant γPKC forms soluble oligomers that have amyloid structure and induce improper development of PC dendrites (20).…”

Section: Introductionmentioning

confidence: 99%

“…Furthermore, mutant γPKC forms soluble oligomers that have amyloid structure and induce improper development of PC dendrites (20). Amyloid oligomers and fibrils caused by mutant or misfolded proteins are also Tanaka 1 frequently observed in various neurodegenerative diseases, including Alzheimer's, Parkinson's, and polyglutamine diseases (22,23), which suggests that aggregation is a common component of the pathogenesis of neurodegenerative diseases like SCA14.…”

Section: Introductionmentioning

confidence: 99%

Congo Red, an Amyloid-Inhibiting Compound, Alleviates Various Types of Cellular Dysfunction Triggered by Mutant Protein Kinase Cγ That Causes Spinocerebellar Ataxia Type 14 (SCA14) by Inhibiting Oligomerization and Aggregation

et al. 2010

Self Cite

View full text Add to dashboard Cite

Abstract. Several missense mutations in the protein kinase Cγ (γPKC) gene have been found to cause spinocerebellar ataxia type 14 (SCA14), an autosomal dominant neurodegenerative disease. We previously demonstrated that the mutant γPKC found in SCA14 is susceptible to aggregation that induces apoptotic cell death. Congo red is widely used as a histological dye for amyloid detection. Recent evidence has revealed that Congo red has the property to inhibit amyloid oligomers and fibril formation of misfolded proteins. In the present study, we examine whether Congo red inhibits aggregate formation and cytotoxicity of mutant γPKC. Congo red likely inhibits aggregate formation of mutant γPKC -green fluorescent protein (GFP) without affecting its expression level in SH-SY5Y cells. Congo red counteracts the insolubilization of recombinant mutant γPKC, suggesting that the dye inhibits aggregation of mutant γPKC by a direct mechanism. Congo red also inhibits aggregation and oligomerization of mutant γPKC-GFP in primary cultured cerebellar Purkinje cells. Moreover, the dye reverses the improper development of dendrites and inhibits apoptotic cell death in Purkinje cells that express mutant γPKC-GFP. These results indicate that amyloid-inhibiting compounds like Congo red may be novel therapeutics for SCA14.

show abstract

Autosomal Dominant Spinocerebellar Ataxias and Episodic Ataxias

Taroni

Chiapparini

Mariotti

2021

Handbook of the Cerebellum and Cerebellar Disorders

View full text Add to dashboard Cite

Mutant γPKC found in spinocerebellar ataxia type 14 induces aggregate-independent maldevelopment of dendrites in primary cultured Purkinje cells

Cited by 58 publications

References 63 publications

Effects of the Chemical Chaperone 4-Phenylbutylate on the Function of the Serotonin Transporter (SERT) Expressed in COS-7 Cells

Effects of the Chemical Chaperone 4-Phenylbutylate on the Function of the Serotonin Transporter (SERT) Expressed in COS-7 Cells

Congo Red, an Amyloid-Inhibiting Compound, Alleviates Various Types of Cellular Dysfunction Triggered by Mutant Protein Kinase Cγ That Causes Spinocerebellar Ataxia Type 14 (SCA14) by Inhibiting Oligomerization and Aggregation

Autosomal Dominant Spinocerebellar Ataxias and Episodic Ataxias

Contact Info

Product

Resources

About