Dimethyl-sulfoxide is a Suitable Solvent for Fluorescent Microscopy Detection of Medium and Strong Heat Shock Inductors Using Transgenic Zebrafish

Heat shock inductors have potential as treatment for degenerative and protein misfolding diseases. Dimethyl-sulfoxide is widely used as a solvent in pharmacological screening tests and has been shown to have heat shock induction effects. Transgenic Tg (hsp70l:EGFP-HRAS_G12V)io3(AB) zebrafish larvae were exposed for 24 hours to dimethyl-sulfoxide in concentratios of 0.1-2%, and to moderate heat shock inductors pentoxifylline and tacrolimus. Positive controls were exposed to 35, 38 and 40�C for 20 min, and incubated for 24 h at 28�C. Heat shock response was measured by fluorescence microscopy and signal intensity quantification in FIJI. Dimethyl-sulfoxide caused a dose-dependant increase in fluorescent intensity, but significantly lower compared with exposure to 38 and 40�C. Pentoxifylline and tacrolimus induced a significantly higher increase in fluorescence compared with 0.5% dimethyl-sulfoxide. Thus, although dimethyl-sulfoxide has independent heat shock induction effects, concentrations of up to 0.5% are suitable for heat shock response screening tests.

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Dynamics of Heat Shock Detection and Response in the Intestine of Caenorhabditis elegans

10.1101/794800 ◽

2019 ◽

Cited By ~ 1

Author(s):

Erin K. Dahlstrom ◽

Erel Levine

Keyword(s):

Caenorhabditis Elegans ◽

Heat Shock ◽

Heat Shock Response ◽

Protein Misfolding ◽

Single Cells ◽

Molecular Response ◽

Longitudinal Measurements ◽

Cell Tissue ◽

Shock Response ◽

Protein Misfolding And Aggregation

AbstractThe heat shock response is the organized molecular response to stressors which disrupt proteostasis, potentially leading to protein misfolding and aggregation. While the regulation of the heat shock response is well-studied in single cells, its coordination at the cell, tissue, and systemic levels of a multicellular organism is poorly understood. To probe the interplay between systemic and cell-autonomous responses, we studied the upregulation of HSP-16.2, a molecular chaperone induced throughout the intestine of Caenorhabditis elegans following a heat shock, by taking longitudinal measurements in a microfluidic environment. Based on the dynamics of HSP-16.2 accumulation, we showed that a combination of heat shock temperature and duration define the intensity of stress inflicted on the worm and identified two regimes of low and high intensity stress. Modeling the underlying regulatory dynamics implicated the saturation of heat shock protein mRNA production in defining these two regimes and emphasized the importance of time separation between transcription and translation in establishing these dynamics. By applying a heat shock and measuring the response in separate parts of the animals, we implicated thermosensory neurons in accelerating the response and transducing information within the animal. We discuss possible implications of the systemic and cell level aspects and how they coordinate to facilitate the organismal response.

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Heat Shock Proteome of Agrobacterium tumefaciens: Evidence for New Control Systems

Journal of Bacteriology ◽

10.1128/jb.184.6.1772-1778.2002 ◽

2002 ◽

Vol 184 (6) ◽

pp. 1772-1778 ◽

Cited By ~ 29

Author(s):

Ran Rosen ◽

Knut Büttner ◽

Dörte Becher ◽

Kenji Nakahigashi ◽

Takashi Yura ◽

...

Keyword(s):

Agrobacterium Tumefaciens ◽

Heat Shock ◽

Heat Shock Proteins ◽

Control Systems ◽

Heat Shock Response ◽

Proteome Analysis ◽

Regulatory Factors ◽

Shock Response ◽

Shock Proteins ◽

Heat Shock Induction

ABSTRACT The regulation of Agrobacterium tumefaciens heat shock genes involves a transcriptional activator (RpoH) and repressor elements (HrcA-CIRCE). Using proteome analysis and mutants in these control elements, we show that the heat shock induction of 32 (out of 56) heat shock proteins is independent of RpoH and HrcA. These results indicate the existence of additional regulatory factors in the A. tumefaciens heat shock response.

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