In Vivo Profiling Reveals a Competent Heat Shock Response in Adult Neurons: Implications for Neurodegenerative Disorders - PubMed (original) (raw)
In Vivo Profiling Reveals a Competent Heat Shock Response in Adult Neurons: Implications for Neurodegenerative Disorders
Alisia Carnemolla et al. PLoS One. 2015.
Abstract
The heat shock response (HSR) is the main pathway used by cells to counteract proteotoxicity. The inability of differentiated neurons to induce an HSR has been documented in primary neuronal cultures and has been proposed to play a critical role in ageing and neurodegeneration. However, this accepted dogma has not been demonstrated in vivo. We used BAC transgenic mice generated by the Gene Expression Nervous System Atlas project to investigate the capacity of striatal medium sized spiny neurons to induce an HSR as compared to that of astrocytes and oligodendrocytes. We found that all cell populations were competent to induce an HSR upon HSP90 inhibition. We also show the presence and relative abundance of heat shock-related genes and proteins in these striatal cell populations. The identification of a competent HSR in adult neurons supports the development of therapeutics that target the HSR pathway as treatments for neurodegenerative disorders.
Conflict of interest statement
Competing Interests: Saliha Moussaoui was an employee of Novartis and is now an employee of Rhenovia Pharma. This does not alter the authors’ adherence to PLOS ONE policies on sharing data and materials.
Figures
Fig 1. mRNA expression level of HS related genes in different striatal cell populations.
Striatal cells were isolated from wild type and transgenic mice at 6 weeks of age 2 hours after treatment with HSP990 (12 mg/kg) or vehicle and sorted based on Gfp expression. RT-qPCR analysis of the expression levels of (A) Hspa1a/b, Dnajb1 and Hspb1 (B) Hsf1 and Sirt1 and (C) Hsp90aa1 and Hsp90ab1 in GFP+ cells isolated from mice treated with HSP990 as compared to those treated with vehicle.
Fig 2. Validation of the purity of the cell populations used for the mRNA analysis Striatal cells were isolated from wild type and transgenic mice at 6 weeks of age 2 hours after treatment with HSP990 (12 mg/kg) or vehicle and sorted based on Gfp expression.
RT-qPCR analysis of the expression levels of (A) Slc12a5, Drd1a, Drd2, Aldh1l1, Gfap and Mbp and (B) Gfp in GFP+ cells isolated from mice treated with HSP990 or vehicle.
Fig 3. Expression of the heat shock proteins and their regulators in the striatal cell populations Striatal cells were isolated from wild type and transgenic mice at 6 weeks of age 2 hours after treatment with HSP990 (12 mg/kg) or vehicle and sorted based on GFP expression.
Western blot analysis of the expression levels of (A) SIRT1, HSP90, HSF1, HSP70 and HSP40 and (B) NEUN, GFAP and GFP in GFP+ cells isolated from mice treated with HSP990 as compared to those treated with vehicle. Loading controls were ATP5B and GAPDH.
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