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Characterization of Muscular Strength of C.

elegans Mutants used as Model


Organism for Neurodegenerative Diseases using Nemaflex.
Simran Singh , Mizanur Rahman , and Siva A. Vanapalli
1

Dept. of Chemistry & Biochemistry, Texas Tech University/ 2Dept. Of Chemical Engineering, Texas Tech University

Introduction

Method: Nemaflex Device

Results

Neurodegenerative Diseases
Alzheimers, Parkinson's, and Huntington
Progressive loss of neuronal function/muscular
dystrophy/problem in coordination.
Neurodegenerative diseases occur because of
mitochondrial dysfunction
oxidative stress, and
protein aggregation.

Figure 3: Effect of genetic interventions on muscular strength. (a) Comparison of force


distribution for the wild-type (n=20) and two of C. elegans muscle mutants, unc-52 (n=15) and
unc-112 (n=20). The inset shows the GFP-labeled sarcomere units for wild-type (N2 bristol),
unc-52 and unc-112.

Alzheimer: beta amyloid peptide aggregation


Parkinson: alpha-synuclein polypeptide aggregation
Huntington: polyglutamine expansions

Why is C. elegans a model organism?


Easily cultured and maintained in lab
Well defined cell lineage and anatomy
Transparency throughout its life cycle
Well established genetics
Contains orthologs and homologs to human genes: notably
dnj-27/ERdj5 ortholog

Animal Model Used for Parkinsons Disease

Figure 1: Microfluidic apparatus for recording muscular force production in C. elegans. (a) Schematic showing the
apparatus configuration including the chamber for housing worms, deformable pillar arrays, microscope objective for
visualizing pillar deflection and locomotion of C. elegans. The housing chamber is made of a non-permeable elastomeric
substrate (Polydimethylsiloxane) for experiments. (b) Schematic of a deflected pillar/sensor by section of worm body
(exaggerated image). (c) Image of a C. elegans in the microfluidic pillar apparatus deforming the sensors. (d) Scanning
electron microscope (SEM) pictures of the pillars.

Method: Procedure for Force Measurement

Figure 4: Effect of cholinergic agonist levamisole (1mM) on muscle force production. (a)
Force production by C. elegans before treatment with drug (red: N =20, n = 3992), during
treatment with drug (green, N = 17, n = 3592) and during recovery after 4 hrs with food (black:
N = 14, n = 3142). Inset is comparison of median force and 95 percentile for each case. (b) C.
elegans before exposure, during exposure and during recovery after 4 hours.

Mutant in this experiment is cc85


Genotype:
unc-119 (ed3) III; pkls2386 [Punc-54::alpha-synuclein::YFP,
unc-119(+)] IV
Phenotype:
Reasonably wild type
Progressive paralysis in adulthood
Green protein aggregates

Figure 2: Muscle force production in wild-type C. elegans (a) Images of the worm interacting with different pillars during
a locomotory episode. Maximum force is experienced by pillar 4 in the first image and by pillar 2 in the second and third
images. The dashed horizontal lines help easy visualization of the pillar deflection. Scale bar is 100 m. (b) Force profiles of
four pillars that the worm is interacting with during the episode. The arrows indicate the maximum force in frames (i)-(iii).
(c) Force signature of a worm constructed by picking up only the maximum force experienced (among all pillars in
interaction) at each frame. Colors represent the contribution of the pillars in construction of the force signature in each frame.
(d) A typical representation of the force data in terms of cumulative probability distribution. (e) A flow chart of cumulative
distribution function estimation.
Muoz-Lobato et. Al. Antioxidants & Redox Signaling,
20(2), 2014

Figure 5: Muscle force production of Parkinsons model. Force production by wild type C.
elegans (red: N =20, n = 3992) and Parkinsons model unc-119 (ed3) III; pkls2386 [Punc54::alpha-synuclein::YFP, unc-119(+)] IV as a model organism for Parkinson's disease (green,
N = 21, n = 2483).

Conclusion
The Nemaflex device can reliably evaluate the strength of C.
elegans/mutants.
The protein aggregation in C. elegans Parkinsons models
produces significantly lower forces.