An Assay to Study Effect of Test Compound against PolyQ-Mediated Neurotoxicity in Worms

Published: April 30, 2023

Abstract

Source: Wang, Q. et al., Caenorhabditis elegans as a Model System for Discovering Bioactive Compounds Against Polyglutamine-Mediated Neurotoxicity. J. Vis. Exp. (2021)

In this video, we describe a method to evaluate the neuroprotective effect of a test compound, astragalan, on neuronal survival in Caenorhabditis elegans from polyQ-mediated neurotoxicity. The assay uses Caenorhabditis elegans expressing the abnormally expanded polyQ repeats fused to GFP in their ASH neurons to measure neuronal survival following test compound treatment.

Protocol

1. PolyQ-mediated neurotoxicity assay Treatment of C. elegans with test samples To prepare nematodes for the polyQ neurotoxicity assay, transfer 300-500 synchronized L1 larvae of HA759 to each well of a 48-well plate with 500 µL of S medium containing OP50 (OD570 of 0.7-0.8) and 5 mg/mL of astragalan; typically three replicate wells for each treatment. Seal the plate with parafilm and incubate at 15 °C and 120 rpm for 3 days. <…

Açıklamalar

The authors have nothing to disclose.

Materials

C. elegans strain
HA759 rtIs11 [osm-10p::GFP + osm-10p::HtnQ150 + dpy-20(+)] Caenorhabditis Genetics Center (CGC) https://cgc.umn.edu/strain/HA759
E. coli strain
OP50 Caenorhabditis Genetics Center (CGC) https://cgc.umn.edu/strain/OP50
Reagent
Agarose Biowest 111860
Sodium azide Sinopharm Chemical Reagent Co., Ltd. 80115560 https://www.reagent.com.cn/goodsDetail/5e981aa807664e26af
551e96ff5f07cd
Equipment
48-well cell culture plate Nest Biotechnology Co., Ltd. 748001 https://www.cell-nest.com/page94?_l=en&product_id=85
Fluorescence microscope Guangzhou Micro-shot Optical Technology Co., Ltd. Mshot MF31-LED https://www.mshot.com/article/442.html
High-content imaging system Molecular Devices ImageXpress Pico https://www.moleculardevices.com/products/cellular-imaging-systems#High-Content-Imaging
Microcentrifuge GeneCompany GENESPEED X1 https://www.genecompany.com/index.php/Home/Goods/goodsdetails/gid/189.html
Microscope digital camera Guangzhou Micro-shot Optical Technology Co., Ltd. MS60 https://www.mshot.com/article/677.html
Microwave Midea Corp.  M1-211A https://www.midea.cn/10000/10000000001
Parafilm M Sigma-Aldrich P7793-1EA https://www.sigmaaldrich.cn/CN/en/product/sigma/p7793?context=product
Software
Image acquisition and analysis software Molecular Devices MetaXpress https://www.moleculardevices.com/ products/cellular-imaging-systems/ acquisition-and-analysis-software/ metaxpress

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Bu Makaleden Alıntı Yapın
An Assay to Study Effect of Test Compound against PolyQ-Mediated Neurotoxicity in Worms. J. Vis. Exp. (Pending Publication), e21285, doi: (2023).

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