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Huntington disease iPSCs show early molecular changes in intracellular signaling, the expression of oxidative stress proteins and the p53 pathway.

Szlachcic WJ, Switonski PM, Krzyzosiak WJ, Figlerowicz M, Figiel M - Dis Model Mech (2015)

Bottom Line: Several molecular and developmental effects of HD have been identified using neural stem cells (NSCs) and differentiated cells, such as neurons and astrocytes.Surprisingly, we found that a number of changes affecting cellular processes in HD were also present in undifferentiated pluripotent HD iPSCs, including the dysregulation of the MAPK and Wnt signaling pathways and the dysregulation of the expression of genes related to oxidative stress, such as Sod1.Interestingly, a common protein interactor of the huntingtin protein and the proteins in the above pathways is p53, and the expression of p53 was dysregulated in HD YAC128 iPSCs and human HD iPSCs.

View Article: PubMed Central - PubMed

Affiliation: Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14, Poznań 61-704, Poland.

No MeSH data available.


Related in: MedlinePlus

SOD1 was overexpressed in the YAC128 iPSCs. (A) Real-time qPCR analysis of the expression of oxidative-stress-related genes revealed the increased expression of Sod1 and less significant differences in the levels of Gpx1 and Prdx1 mRNAs. (B,C) Western blotting analyses of several clonal lines revealed the increased expression of SOD1 protein but not PRDX1 protein in HD YAC128 iPSCs. (B) Notably, additional SOD1-immunoreactive bands of approximately 29 and 36 kDa were observed in HD YAC128 iPSCs but not in WT iPSCs. * indicates the YAC128/Oct-eGFP lines. (D) Increased expression of SOD1 protein was also observed in the human HD fibroblast line GM04281 (Coriell) using western blotting.
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DMM019406F5: SOD1 was overexpressed in the YAC128 iPSCs. (A) Real-time qPCR analysis of the expression of oxidative-stress-related genes revealed the increased expression of Sod1 and less significant differences in the levels of Gpx1 and Prdx1 mRNAs. (B,C) Western blotting analyses of several clonal lines revealed the increased expression of SOD1 protein but not PRDX1 protein in HD YAC128 iPSCs. (B) Notably, additional SOD1-immunoreactive bands of approximately 29 and 36 kDa were observed in HD YAC128 iPSCs but not in WT iPSCs. * indicates the YAC128/Oct-eGFP lines. (D) Increased expression of SOD1 protein was also observed in the human HD fibroblast line GM04281 (Coriell) using western blotting.

Mentions: HD is associated with the increased production of free radicals and the resultant oxidative stress, which leads to neuronal dysfunction (Hands et al., 2011; Valencia et al., 2013). Antioxidant-related proteins defend cells from oxidative stress and free radicals. To determine whether the expression of such proteins was induced in early undifferentiated HD YAC128 iPSCs, we investigated the expression levels of Sod1, Gpx1 and Prdx1. The Sod1 mRNA expression level was increased in the HD YAC128 iPSCs, and the differences found in the expression levels of Gpx1 and Prdx1 genes were less statistically significant (Fig. 5A). Subsequently, we analyzed SOD1 protein expression levels and found that expression of its 16-kDa monomeric form was also increased in the YAC128 iPSCs (Fig. 5B,C). A similar increase in the level of SOD1 expression was observed in the human HD fibroblast cell line (GM04281, Fig. 5D). Interestingly, other SOD1-immunoreactive bands were also selectively present in the protein lysates of the YAC128 iPSCs. We detected SOD1-immunoreactive compounds with apparent molecular weights of approximately 29 and 36 kDa (Fig. 5B). These compounds might represent SOD1 oligomers, and they were virtually undetectable in the WT iPSCs.Fig. 5.


Huntington disease iPSCs show early molecular changes in intracellular signaling, the expression of oxidative stress proteins and the p53 pathway.

Szlachcic WJ, Switonski PM, Krzyzosiak WJ, Figlerowicz M, Figiel M - Dis Model Mech (2015)

SOD1 was overexpressed in the YAC128 iPSCs. (A) Real-time qPCR analysis of the expression of oxidative-stress-related genes revealed the increased expression of Sod1 and less significant differences in the levels of Gpx1 and Prdx1 mRNAs. (B,C) Western blotting analyses of several clonal lines revealed the increased expression of SOD1 protein but not PRDX1 protein in HD YAC128 iPSCs. (B) Notably, additional SOD1-immunoreactive bands of approximately 29 and 36 kDa were observed in HD YAC128 iPSCs but not in WT iPSCs. * indicates the YAC128/Oct-eGFP lines. (D) Increased expression of SOD1 protein was also observed in the human HD fibroblast line GM04281 (Coriell) using western blotting.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4582098&req=5

DMM019406F5: SOD1 was overexpressed in the YAC128 iPSCs. (A) Real-time qPCR analysis of the expression of oxidative-stress-related genes revealed the increased expression of Sod1 and less significant differences in the levels of Gpx1 and Prdx1 mRNAs. (B,C) Western blotting analyses of several clonal lines revealed the increased expression of SOD1 protein but not PRDX1 protein in HD YAC128 iPSCs. (B) Notably, additional SOD1-immunoreactive bands of approximately 29 and 36 kDa were observed in HD YAC128 iPSCs but not in WT iPSCs. * indicates the YAC128/Oct-eGFP lines. (D) Increased expression of SOD1 protein was also observed in the human HD fibroblast line GM04281 (Coriell) using western blotting.
Mentions: HD is associated with the increased production of free radicals and the resultant oxidative stress, which leads to neuronal dysfunction (Hands et al., 2011; Valencia et al., 2013). Antioxidant-related proteins defend cells from oxidative stress and free radicals. To determine whether the expression of such proteins was induced in early undifferentiated HD YAC128 iPSCs, we investigated the expression levels of Sod1, Gpx1 and Prdx1. The Sod1 mRNA expression level was increased in the HD YAC128 iPSCs, and the differences found in the expression levels of Gpx1 and Prdx1 genes were less statistically significant (Fig. 5A). Subsequently, we analyzed SOD1 protein expression levels and found that expression of its 16-kDa monomeric form was also increased in the YAC128 iPSCs (Fig. 5B,C). A similar increase in the level of SOD1 expression was observed in the human HD fibroblast cell line (GM04281, Fig. 5D). Interestingly, other SOD1-immunoreactive bands were also selectively present in the protein lysates of the YAC128 iPSCs. We detected SOD1-immunoreactive compounds with apparent molecular weights of approximately 29 and 36 kDa (Fig. 5B). These compounds might represent SOD1 oligomers, and they were virtually undetectable in the WT iPSCs.Fig. 5.

Bottom Line: Several molecular and developmental effects of HD have been identified using neural stem cells (NSCs) and differentiated cells, such as neurons and astrocytes.Surprisingly, we found that a number of changes affecting cellular processes in HD were also present in undifferentiated pluripotent HD iPSCs, including the dysregulation of the MAPK and Wnt signaling pathways and the dysregulation of the expression of genes related to oxidative stress, such as Sod1.Interestingly, a common protein interactor of the huntingtin protein and the proteins in the above pathways is p53, and the expression of p53 was dysregulated in HD YAC128 iPSCs and human HD iPSCs.

View Article: PubMed Central - PubMed

Affiliation: Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14, Poznań 61-704, Poland.

No MeSH data available.


Related in: MedlinePlus