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Young adult Caenorhabditis elegans nematodes are exposed to different concentrations of commercial pesticides or other toxicants for 2-24 h. Then, different neurons can be visualized using fluorescent-expressing strains. This paper demonstrates how to expose nematodes to pesticides and assess neuron damage.
Caenorhabditis elegans is a powerful model organism used in many research laboratories to understand the consequences of exposure to chemical pollutants, pesticides, and a wide variety of toxic substances. These nematodes are easy to work with and can be used to generate novel research findings, even in the undergraduate biology laboratory. A multi-week laboratory series of authentic, student-driven research projects trains students in a toolkit of techniques and approaches in behavioral measurements, cell biology, and microscopy that they then apply to their projects. One technique in that toolkit is quantifying the percentage of neurons exhibiting neurodegenerative damage following exposure to a chemical toxicant like a pesticide. Young adult C. elegans nematodes can be exposed to different concentrations of commercially available pesticides or other types of toxicants for 2-24 h. Then, undergraduate students can visualize different neuron subtypes using fluorescent-expressing strains of C. elegans. These techniques do not require sophisticated image processing software and are effective at even low magnifications, making the need for expensive confocal microscopy unnecessary. This paper demonstrates how to treat the nematodes with pesticides and how to image and score the neurons. It also provides a straightforward protocol for the microscopy and analysis of neuron morphology. The materials used for this technique are inexpensive and readily available in most undergraduate biology departments. This technique can be combined with behavioral measures like locomotion, basal slowing, or egg-laying to conduct a potentially publishable series of experiments and give undergraduate students an authentic research experience at a very low cost.
Caenorhabditis elegans is an excellent model organism for laboratory course training in biological science courses for introductory- and intermediate-level students. This laboratory procedure can be used as part of a multi-week module that explores various effects of commonly used pesticides on C. elegans behavior and cell biology. Students can learn how to design and carry out independent projects that teach them data analysis and presentation skills. This paper focuses on the protocols for exposing C. elegans to pesticide mixtures and then observing and analyzing the effects on neuron morphology.
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All use of invertebrate animals was in compliance with the animal care and use guidelines of the institution.
1. Preparation of pesticide-coated Petri plates
The methods and protocols described in this paper provide important laboratory skills for intermediate-level undergraduate students in biology or neuroscience. Students can gain important experience in developing an independent project and conducting an experiment of their own design that could provide novel results. Figure 3 shows an optimal result of a student project that eventually became part of a published paper3. While the majority of student projects do not re.......
The protocols described in this manuscript work successfully alone or as part of a multi-week independent student group project. The protocols are also amenable to stand-alone, one-week exploratory experiences. The nematode strains are cheap and easy to maintain in the research laboratory. Students can readily learn how to pick worms with a worm pick or how to move them by flushing plates with water and allowing them to settle by gravity. The experiments can be performed over the course of a single lab period or over mul.......
The work described in this manuscript was done for an intermediate-level class in neuroscience. Funds for the reagents and supplies were provided by the Biology Department at Vassar College. The microscopes and digital imaging system were also provided by the Biology Department at Vassar College. The author thanks all the many students who took this course.
....Name | Company | Catalog Number | Comments |
Agar | Fisher Scientific | BP97445 | |
Agarose | Fisher Scientific | MP1AGAH0250 | |
Alcohol lamp | Fisher Scientific | 17012826 | |
Bunsen burner | Fisher Scientific | 17-012-820 | |
C. elegans strains | C. elegans Genetics Center | ||
CaCl | Fisher Scientific | 10035-04-8 | |
Cholesterol | Fisher Scientific | AAA1147030 | |
Coverslips | Fisher Scientific | 12-545-AP | |
Digital camera | Nikon | These can vary depending on the requirement | |
Dissecting scope | Nikon | SMZ745 | |
E. coli strain (OP50) | C. elegans Genetics Center | ||
Ethanol | Fisher Scientific | BP2818100 | |
Fluorescent scope | Nikon | These can vary depending on the requirement | |
Imaging software | Nikon | These can vary depending on the requirement | |
Inoculation loop | Fisher Scientific | 131045 | |
LB Broth Base | Fisher Scientific | BP9723-500 | |
MgSO4 | Fisher Scientific | 10034-99-8 | |
Microfuge tubes | Fisher Scientific | 05408129 | |
Microscope slides | Fisher Scientific | 22-265446 | |
Pasteur pipets | Fisher Scientific | 13-678-20A | |
Petri dishes | Fisher Scientific | AS4050 | |
Pipette tips | Fisher Scientific | 94060316 | |
Pipetters | Fisher Scientific | 14-386-319 | |
Platinum wire | Genesee Scientific | 59-1M30P | |
Potassium Phosphate buffer | Fisher Scientific | AAJ61413AP | |
Sodium azide | Fisher Scientific | AC447810250 |
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