Fluorescent Reporter-Based Paralysis Assay: A Technique to Assess Age-Associated Progressive Formation of Polyglutamine Fluorescent Reporter and Associated Paralysis in Caenorhabditis elegans

Abstract

Source: Lazaro-Pena, M. I. et al. Quantifying Tissue-Specific Proteostatic Decline in Caenorhabditis elegans. J. Vis. Exp. (2021)

This video demonstrates an in vivo imaging-based assay to determine proteostasis decline due to aging. The assay uses Caenorhabditis elegans expressing polyglutamine YFP-fusion protein in body wall muscles to measure the age-associated aggregation of polyglutamine and induction of proteotoxicity, leading to paralysis.

Protocol

1. Measuring the decline in proteostasis in muscle tissue by using polyglutamine-expressing animals

NOTE: Two methods can be used to identify proteostasis decline in muscle cells: imaging the formation of protein aggregates during aging (1.1) and measuring the proteotoxicity these aggregates cause with age through the onset of paralysis (1.2).

1. Imaging polyglutamine aggregate formation in the muscle during aging
   NOTE: The age-dependent progression of protein aggregation in the muscle cells is imaged using polyglutamine (polyQ) repeats fused to a yellow fluorescent protein ....

Results

Figure 1: Expression of polyQ::YFP within C. elegans muscle results in progressive foci accumulation and paralysis during aging. (A) C. elegans unc-54p::Q35::YFP expression at days 1 and 4 of adulthood (u.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Glass microscope slides	VWR	160004-422
Sodium Azide, CAS #26628-22-8	Sigma-Aldrich	S2002
Zeiss Axio Imager M2m microscope with AxioVision v4.8.2.0 software	Zeiss
Zeiss StemiSV11 M2 Bio Quad microscope	Zeiss