Introduction to C. elegans Genetic Model System
Instructor: Niall Howlett, PhD, University of Rhode Island
Location: URI
Session 1: June 5 – June 7 ( Lab: Room 405 / Lecture Room: Avedisian Rm 303)
Session 2: June 12 – June 14 ( Lab: Room 405 / Lecture Room: Avedisian Rm 403)
Course Overview
For well over 50 years, since the pioneering work of Sydney Brenner, the nematode (round worm) Caenorhabditis elegans (C. elegans) has been used as a genetic model to study developmental biology and nervous system development and function. C. elegans is also an outstanding model organism for the study of basic functions and interactions of eukaryotic cells, host-parasite interactions, evolution, aging, and disease.
There are many features of C. elegans that make it an outstanding model organism; 1) Its small size (0.25 – 1.0 mm in length), 2) Its rapid life cycle (3 days at 25°C from egg to egg-laying adult | adults live for ~2 weeks), 3) It is transparent, enabling brightfield and fluorescence visualization of development and function, 4) It’s small and fully sequenced genome (100 Mbp) – the first multicellular organism to have its genome sequenced, and 5) It’s powerful genetics, e.g., the ease of genetic manipulation and ability to perform forward and reverse genetic screens to identify genes associated with novel phenotypes.
The module will be completed over 2.5 days and participants who complete the module will receive an RI-INBRE Certificate of completion.
Learning Outcomes
Upon completion of this workshop, we expect that trainees will be able to perform the following:
- Prepare C. elegans growth media including the OP50 bacterial food source.
- Understand the lifecycle of C. elegans from egg to aged adult and be able to recognize the various larval and adult life stages.
- Understand C. elegans gene, protein, and strain nomenclature.
- Recognize key anatomical features of adult C. elegans.
- Physically pick and isolate C. elegans at select larval and adult life stages using the stereomicroscope.
- Perform C. elegans fluorescence microscopy to examine specific neuronal cell populations.
- Perform select C. elegans behavioral assays.
Lab Report
Students will be expected to maintain detailed laboratory notes
Timeline
| Day 1 | Day 2 | Day 3 | |
|---|---|---|---|
| 9:00 AM-10:00 AM | Intro to C. elegans as a model organism (lecture) | Behavioral assays: Measuring worm motor neuron activity using the thrashing assay (lecture) | |
| 10:00 AM-11:00 AM | Identification of the different stages of the life cycle. Noting worm behavior, characteristics, and movement (hands-on) | Behavioral assay: The thrashing assay with wild-type and mutant worms (hands-on) | Practice picking worms, review all collected data (hands on and discussion) |
| 11:00 AM-12:00 PM | Introduction to worm media preparation and the culturing of the bacterial food source using aseptic technique (lecture and hands on) | Analysis of thrashing results; graphing and statistical analysis. Discussion and interpretation of the results | Open discussion and Q&A session |
| 12:00 - 1:00 PM | Break | Break | WDT Survey and Certificate Distribution |
| 1:00 PM-2:00 PM | Completion of C. elegans media preparation. Discussion of C. elegans gene, protein, and strain nomenclature (Hands on & lecture) | The use of GFP transgenic reporter strains for nervous system function (lecture) | |
| 2:00 PM-3:00 PM | Analysis of C. elegans movement and behavior; wild-type and mutant worms, recording observations (hands-on) | Analysis of wild-type and mutant GFP-labelled acetylcholine (ACh) motor neuron reporter strains | Performing neuronal counts (hands-on) | |
| 3:00 PM-4:00 PM | Practice picking and moving worms on the stereomicroscope (hands-on) | Analysis of ACh results; graphing and statistical analysis | Discussion and interpretation of the results |