池田婚姻願（宮内大臣宛様式）

The heat shock response (HSR) is a mechanism to cope with proteotoxic stress by inducing the expression of molecular chaperones and other heat shock response genes. The HSR is evolutionarily well conserved and has been widely studied in bacteria, cell lines and lower eukaryotic model organisms. However, mechanistic insights into the HSR in higher eukaryotes, in particular in mammals, are limited. We have developed an in vivo heat shock protocol to analyze the HSR in mice and dissected heat shock factor 1 (HSF1)-dependent and-independent pathways. Whilst the induction of proteostasis-related genes was dependent on HSF1, the regulation of circadian function related genes, indicating that the circadian clock oscillators have been reset, was independent of its presence. Furthermore, we demonstrate that the in vivo HSR is impaired in mouse models of Huntington's disease but we were unable to corroborate the general repression of transcription that follows a heat shock in lower eukaryotes

Contesting the dogma of an age-related heat shock response impairment:implications for cardiac-specific age-related disorders

Ageing is associated with the reduced performance of physiological processes and has been proposed as a major risk factor for disease. An age-related decline in stress response pathways has been widely documented in lower organisms. In particular, the heat shock response (HSR) becomes severely compromised with age in Caenorhabditis elegans. However, a comprehensive analysis of the consequences of ageing on the HSR in higher organisms has not been documented. We used both HS and inhibition of HSP90 to induce the HSR in wild-type mice at 3 and 22 months of age to investigate the extent to which different brain regions, and peripheral tissues can sustain HSF1 activity and HS protein (HSP) expression with age. Using chromatin immunoprecipitation, quantitative reverse transcription polymerase chain reaction, western blotting and enzyme linked immunosorbent assay (ELISA), we were unable to detect a difference in the level or kinetics of HSP expression between young and old mice in all brain regions. In contrast, we did observe an age-related reduction in chaperone levels and HSR-related proteins in the heart. This could result in a decrease in the protein folding capacity of old hearts with implications for age-related cardiac disorder

<i>In Vivo</i> Profiling Reveals a Competent Heat Shock Response in Adult Neurons: Implications for Neurodegenerative Disorders

<div><p>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 <i>in vivo</i>. 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.</p></div

mRNA expression level of HS related genes in different striatal cell populations.

<p>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 <i>Gfp</i> expression. RT-qPCR analysis of the expression levels of (A) <i>Hspa1a/b</i>, <i>Dnajb1</i> and <i>Hspb1</i> (B) <i>Hsf1</i> and <i>Sirt1</i> and (C) <i>Hsp90aa1</i> and <i>Hsp90ab1</i> in GFP⁺ cells isolated from mice treated with HSP990 as compared to those treated with vehicle.</p

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.

<p>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.</p

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 <i>Gfp</i> expression.

<p>RT-qPCR analysis of the expression levels of (A) <i>Slc12a5</i>, <i>Drd1a</i>, <i>Drd2</i>, <i>Aldh1l1</i>, <i>Gfap</i> and <i>Mbp</i> and (B) <i>Gfp</i> in GFP⁺ cells isolated from mice treated with HSP990 or vehicle.</p

Myostatin inhibition prevents skeletal muscle pathophysiology in Huntington's disease mice

Huntington’s disease (HD) is an inherited neurodegenerative disorder of which skeletal muscle atrophy is a common feature, and multiple lines of evidence support a muscle-based pathophysiology in HD mouse models. Inhibition of myostatin signaling increases muscle mass, and therapeutic approaches based on this are in clinical development. We have used a soluble ActRIIB decoy receptor (ACVR2B/Fc) to test the effects of myostatin/activin A inhibition in the R6/2 mouse model of HD. Weekly administration from 5 to 11 weeks of age prevented body weight loss, skeletal muscle atrophy, muscle weakness, contractile abnormalities, the loss of functional motor units in EDL muscles and delayed end-stage disease. Inhibition of myostatin/activin A signaling activated transcriptional profiles to increase muscle mass in wild type and R6/2 mice but did little to modulate the extensive Huntington’s disease-associated transcriptional dysregulation, consistent with treatment having little impact on HTT aggregation levels. Modalities that inhibit myostatin signaling are currently in clinical trials for a variety of indications, the outcomes of which will present the opportunity to assess the potential benefits of targeting this pathway in HD patients