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Indole and synthetic derivative activate chaperone expression to reduce polyQ aggregation in SCA17 neuronal cell and slice culture models.

Kung PJ, Tao YC, Hsu HC, Chen WL, Lin TH, Janreddy D, Yao CF, Chang KH, Lin JY, Su MT, Wu CH, Lee-Chen GJ, Hsieh-Li HM - Drug Des Devel Ther (2014)

Bottom Line: The molecular chaperones act in preventing protein aggregation to ameliorate downstream harmful events.We found that indole and NC001-8 up-regulated chaperone expression to reduce polyQ aggregation in neuronal differentiated TBP/Q79 cells.Our results demonstrate how indole and derivative NC001-8 reduce polyQ aggregation to support their therapeutic potentials in SCA17 treatment.

View Article: PubMed Central - PubMed

Affiliation: Department of Life Science, National Taiwan Normal University, Taipei, Taiwan.

ABSTRACT
In spinocerebellar ataxia type 17 (SCA17), the expansion of a translated CAG repeat in the TATA box binding protein (TBP) gene results in a long polyglutamine (polyQ) tract in the TBP protein, leading to intracellular accumulation of aggregated TBP and cell death. The molecular chaperones act in preventing protein aggregation to ameliorate downstream harmful events. In this study, we used Tet-On SH-SY5Y cells with inducible SCA17 TBP/Q79-green fluorescent protein (GFP) expression to test indole and synthetic derivative NC001-8 for neuroprotection. We found that indole and NC001-8 up-regulated chaperone expression to reduce polyQ aggregation in neuronal differentiated TBP/Q79 cells. The effects on promoting neurite outgrowth and on reduction of aggregation on Purkinje cells were also confirmed with cerebellar primary and slice cultures of SCA17 transgenic mice. Our results demonstrate how indole and derivative NC001-8 reduce polyQ aggregation to support their therapeutic potentials in SCA17 treatment.

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Indole and NC001-8 promoted neurite outgrowth and reduced aggregation of Purkinje cells in spinocerebellar ataxia type 17 mouse cerebellar primary and slice cultures.Notes: The primary culture was treated with 0~100 nM indole (A) or NC001-8 (B) for 13 days. The representative microscopic images of treatment with 100 nM indole or NC001-8 are shown, and the relative Purkinje cell neurite outgrowth (green) and aggregation (shown in white in the middle column and in red in the right merged column) were quantified (n=3). (C) The slice culture was treated with 10 nM indole or 10 μM NC001-8 for 6 days. The representative microscopic images of treatment are shown, and the relative Purkinje cell aggregation (red) was quantified (n=3). To normalize, the relative neurite outgrowth length and aggregation level in vehicle-treated cells or slices is set as 100%. IP3R-1 and 1TBP18 antibodies were used to detect the Purkinje cells and TBP aggregation, respectively.Abbreviations: IP3R-1, inositol 1,4,5 trisphosphate receptor type 1; 1TBP18, mouse monoclonal antibody to TATA binding protein (TBP); Rel., relative; DAPI, 4′,6-diamidino-2-phenylindole.
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f5-dddt-8-1929: Indole and NC001-8 promoted neurite outgrowth and reduced aggregation of Purkinje cells in spinocerebellar ataxia type 17 mouse cerebellar primary and slice cultures.Notes: The primary culture was treated with 0~100 nM indole (A) or NC001-8 (B) for 13 days. The representative microscopic images of treatment with 100 nM indole or NC001-8 are shown, and the relative Purkinje cell neurite outgrowth (green) and aggregation (shown in white in the middle column and in red in the right merged column) were quantified (n=3). (C) The slice culture was treated with 10 nM indole or 10 μM NC001-8 for 6 days. The representative microscopic images of treatment are shown, and the relative Purkinje cell aggregation (red) was quantified (n=3). To normalize, the relative neurite outgrowth length and aggregation level in vehicle-treated cells or slices is set as 100%. IP3R-1 and 1TBP18 antibodies were used to detect the Purkinje cells and TBP aggregation, respectively.Abbreviations: IP3R-1, inositol 1,4,5 trisphosphate receptor type 1; 1TBP18, mouse monoclonal antibody to TATA binding protein (TBP); Rel., relative; DAPI, 4′,6-diamidino-2-phenylindole.

Mentions: To further confirm the neuroprotective potential of the indole compounds, we applied indole and NC001-8 to the cerebellar primary and slice cultures established from SCA17 mice.34 As shown in Figure 5A-B, with 10~100 nM compound concentration, significantly (indole: 134%~149%; P=0.016~0.002) or notably (NC001-8: 114%~119%; P>0.05) increased Purkinje cell neurite outgrowth was observed. Both compounds at concentrations 1~100 nM significantly reduced the Purkinje cell aggregation on the primary culture (indole: P=0.039~0.001; NC001-8: P=0.042~0.016). The percentage of cells with aggregates under indole treatment is 86% (1 nM), 71% (10 nM), and 53% (100 nM) in Figure 5A, and the percentage of cells with aggregates under NC001-8 treatment is 60% (1 nM), 68% (10 nM), and 60% (100 nM) in Figure 5B. On SCA17 mouse cerebellar slice culture, although indole at 10 nM could significantly reduce the Purkinje cell aggregation (28%; P=0.001), 1,000 folds of NC001-8 (10 μM) were required to obtain a significant reduction of the aggregation (16%; P<0.001) (Figure 5C). Thus, indole worked more efficiently than NC001-8 in reducing the Purkinje cell aggregation on SCA17 mouse cerebellar slice culture.


Indole and synthetic derivative activate chaperone expression to reduce polyQ aggregation in SCA17 neuronal cell and slice culture models.

Kung PJ, Tao YC, Hsu HC, Chen WL, Lin TH, Janreddy D, Yao CF, Chang KH, Lin JY, Su MT, Wu CH, Lee-Chen GJ, Hsieh-Li HM - Drug Des Devel Ther (2014)

Indole and NC001-8 promoted neurite outgrowth and reduced aggregation of Purkinje cells in spinocerebellar ataxia type 17 mouse cerebellar primary and slice cultures.Notes: The primary culture was treated with 0~100 nM indole (A) or NC001-8 (B) for 13 days. The representative microscopic images of treatment with 100 nM indole or NC001-8 are shown, and the relative Purkinje cell neurite outgrowth (green) and aggregation (shown in white in the middle column and in red in the right merged column) were quantified (n=3). (C) The slice culture was treated with 10 nM indole or 10 μM NC001-8 for 6 days. The representative microscopic images of treatment are shown, and the relative Purkinje cell aggregation (red) was quantified (n=3). To normalize, the relative neurite outgrowth length and aggregation level in vehicle-treated cells or slices is set as 100%. IP3R-1 and 1TBP18 antibodies were used to detect the Purkinje cells and TBP aggregation, respectively.Abbreviations: IP3R-1, inositol 1,4,5 trisphosphate receptor type 1; 1TBP18, mouse monoclonal antibody to TATA binding protein (TBP); Rel., relative; DAPI, 4′,6-diamidino-2-phenylindole.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4206201&req=5

f5-dddt-8-1929: Indole and NC001-8 promoted neurite outgrowth and reduced aggregation of Purkinje cells in spinocerebellar ataxia type 17 mouse cerebellar primary and slice cultures.Notes: The primary culture was treated with 0~100 nM indole (A) or NC001-8 (B) for 13 days. The representative microscopic images of treatment with 100 nM indole or NC001-8 are shown, and the relative Purkinje cell neurite outgrowth (green) and aggregation (shown in white in the middle column and in red in the right merged column) were quantified (n=3). (C) The slice culture was treated with 10 nM indole or 10 μM NC001-8 for 6 days. The representative microscopic images of treatment are shown, and the relative Purkinje cell aggregation (red) was quantified (n=3). To normalize, the relative neurite outgrowth length and aggregation level in vehicle-treated cells or slices is set as 100%. IP3R-1 and 1TBP18 antibodies were used to detect the Purkinje cells and TBP aggregation, respectively.Abbreviations: IP3R-1, inositol 1,4,5 trisphosphate receptor type 1; 1TBP18, mouse monoclonal antibody to TATA binding protein (TBP); Rel., relative; DAPI, 4′,6-diamidino-2-phenylindole.
Mentions: To further confirm the neuroprotective potential of the indole compounds, we applied indole and NC001-8 to the cerebellar primary and slice cultures established from SCA17 mice.34 As shown in Figure 5A-B, with 10~100 nM compound concentration, significantly (indole: 134%~149%; P=0.016~0.002) or notably (NC001-8: 114%~119%; P>0.05) increased Purkinje cell neurite outgrowth was observed. Both compounds at concentrations 1~100 nM significantly reduced the Purkinje cell aggregation on the primary culture (indole: P=0.039~0.001; NC001-8: P=0.042~0.016). The percentage of cells with aggregates under indole treatment is 86% (1 nM), 71% (10 nM), and 53% (100 nM) in Figure 5A, and the percentage of cells with aggregates under NC001-8 treatment is 60% (1 nM), 68% (10 nM), and 60% (100 nM) in Figure 5B. On SCA17 mouse cerebellar slice culture, although indole at 10 nM could significantly reduce the Purkinje cell aggregation (28%; P=0.001), 1,000 folds of NC001-8 (10 μM) were required to obtain a significant reduction of the aggregation (16%; P<0.001) (Figure 5C). Thus, indole worked more efficiently than NC001-8 in reducing the Purkinje cell aggregation on SCA17 mouse cerebellar slice culture.

Bottom Line: The molecular chaperones act in preventing protein aggregation to ameliorate downstream harmful events.We found that indole and NC001-8 up-regulated chaperone expression to reduce polyQ aggregation in neuronal differentiated TBP/Q79 cells.Our results demonstrate how indole and derivative NC001-8 reduce polyQ aggregation to support their therapeutic potentials in SCA17 treatment.

View Article: PubMed Central - PubMed

Affiliation: Department of Life Science, National Taiwan Normal University, Taipei, Taiwan.

ABSTRACT
In spinocerebellar ataxia type 17 (SCA17), the expansion of a translated CAG repeat in the TATA box binding protein (TBP) gene results in a long polyglutamine (polyQ) tract in the TBP protein, leading to intracellular accumulation of aggregated TBP and cell death. The molecular chaperones act in preventing protein aggregation to ameliorate downstream harmful events. In this study, we used Tet-On SH-SY5Y cells with inducible SCA17 TBP/Q79-green fluorescent protein (GFP) expression to test indole and synthetic derivative NC001-8 for neuroprotection. We found that indole and NC001-8 up-regulated chaperone expression to reduce polyQ aggregation in neuronal differentiated TBP/Q79 cells. The effects on promoting neurite outgrowth and on reduction of aggregation on Purkinje cells were also confirmed with cerebellar primary and slice cultures of SCA17 transgenic mice. Our results demonstrate how indole and derivative NC001-8 reduce polyQ aggregation to support their therapeutic potentials in SCA17 treatment.

Show MeSH
Related in: MedlinePlus