Open AccessMeasurements of Physiological Stress Responses in <em>C. Elegans</em>
Author(s) -
Raz BarZiv,
Ashley E. Frakes,
Ryo HiguchiSanabria,
Theodore Bolas,
Phillip A. Frankino,
Holly K. Gildea,
Melissa G. Metcalf,
Andrew Dillin
Publication year - 2020
Publication title -
journal of visualized experiments
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.596
H-Index - 91
ISSN - 1940-087X
DOI - 10.3791/61001
Subject(s) - unfolded protein response , proteostasis , microbiology and biotechnology , biology , endoplasmic reticulum , cellular stress response , caenorhabditis elegans , heat shock , proteome , mitochondrion , proteotoxicity , heat shock protein , homeostasis , fight or flight response , protein folding , bioinformatics , genetics , gene
Organisms are often exposed to fluctuating environments and changes in intracellular homeostasis, which can have detrimental effects on their proteome and physiology. Thus, organisms have evolved targeted and specific stress responses dedicated to repair damage and maintain homeostasis. These mechanisms include the unfolded protein response of the endoplasmic reticulum (UPR ER ), the unfolded protein response of the mitochondria (UPR MT ), the heat shock response (HSR), and the oxidative stress response (OxSR). The protocols presented here describe methods to detect and characterize the activation of these pathways and their physiological consequences in the nematode, C. elegans. First, the use of pathway-specific fluorescent transcriptional reporters is described for rapid cellular characterization, drug screening, or large-scale genetic screening (e.g., RNAi or mutant libraries). In addition, complementary, robust physiological assays are described, which can be used to directly assess sensitivity of animals to specific stressors, serving as functional validation of the transcriptional reporters. Together, these methods allow for rapid characterization of the cellular and physiological effects of internal and external proteotoxic perturbations.