Neurodegeneration

The assay to monitor the effects of test compounds on aggregate formation and aggregation dynamics in numerous C. elegans models of neurodegenerative diseases.

Method description

For this assay we take advantage of the existing C. elegans GFP-reporter strains, modelling a variety of neurodegenerative disorders, such as Alzheimer’s, Parkinson’s, Huntington’s disease and amyotrophic lateral sclerosis (ALS), as well as the capacity of our device to perform fluorescent imaging.

A synchronized population of C. elegans is injected into the microfluidic platform at the first larval stage (L1). Worms are then confined within dedicated microfluidic chambers and are continuously fed with an E. coli solution.

Worms are chronically exposed to the test compounds starting from the last larval stage prior to sexual maturity (L4) for 85 hours (day 3 of adulthood). The compounds to be tested are mixed with the E. coli solution. Freeze-dried OP50 E. coli are used as a food source for the whole duration of the experiment, preventing the metabolization of the tested molecules by the bacteria.

The images of each microfluidic chamber are recorded every hour. Starting from 70 hours post worm injection (day 1 of adulthood), both brightfield and fluorescent images start to be recorded in order to monitor worm growth and protein aggregation dynamics in parallel. Time-resolved phenotypic readouts are then extracted from the collected images in both brightfield and fluorescent modes.

Readouts

  • Aggregate number
  • Aggregate size

Dynamics of polyQ::yfp aggregation in an AM140 humanized worm (C. elegans model for human Huntington’s disease), as observed via fluorescence imaging by the SydLab system.

Protein aggregation (count)

Temporal evolution of the average number of polyQ::yfp aggregates counted per each worm of two different strains: AM140 (C. elegans model for human Huntington’s disease) and AM134 (control strain). “Humanized” C. elegans strains modeling all major neurodegenerative disorders exist, also including e.g. ALS, Alzheimer’s, and Parkinson’s disease.

Protein aggregation (size)

Temporal evolution of the average size of polyQ::yfp aggregates counted per each worm of two different strains: AM140 (C. elegans model for human Huntington’s disease) and AM134 (control strain). “Humanized” C. elegans strains modeling all major neurodegenerative disorders exist, also including e.g. ALS, Alzheimer’s, and Parkinson’s disease.

Assay combinations

The neurodegeneration assay can be offered on its own or in combination with the following assays upon request.