-
[
International C. elegans Meeting,
2001]
I have used C. elegans in two different undergraduate lab courses at Santa Clara University. One is an upper division research-focused "Project Lab" course described below and the other is an upper division Genetics course that uses C. elegans for its laboratory component. The "Project Lab" course was developed with an NSF grant to integrate research and education by designing and implementing an intensive new laboratory course in which undergraduate students participate in supervised research projects. These research projects are directly related to ongoing research in my laboratory. Project Lab has a class size of 7-12 students and, each year, it focuses on research activities appropriate for that particular stage of the research project. The addition of this course to our biology curriculum enhances the science education of undergraduate students at Santa Clara University by providing hands-on laboratory experience and exposure to real research problems. It also enhances the research program of the principal investigator by training and motivating students for research. Specifically, we are using sequencing and site-directed mutagenesis to conduct a structure/function analysis of LIN-31, a member of the winged-helix family of transcription factors, which is required for the proper specification of vulval cell fates in C. elegans . The first two Project Lab classes made considerable progress, resulting in a paper on which three Project Lab students were authors (Miller et al. , Genetics 156: 1595, 2000). Results from the two most recent Project Lab class will be presented as a poster at this meeting (see Mora-Blanco et al. ). I have also successfully used C. elegans in my upper division genetics laboratory course. This is a 10-week course in which students use C. elegans to study basic genetic principles such as dominance, independent assortment, recombination, and complementation. In order to do this, they first find and then analyze their own genetic mutant. As the quarter progresses, they use their mutants to study the basic genetic principles listed above. The first lab introduces them to C. elegans and teaches them how to grow, manipulate, and sex the animals. In the second lab, they "find" their mutant by doing a mock mutant screen. In subsequent labs, they carry out genetic manipulations (crosses) to better understand their mutants. By the end of the quarter, they can answer the following questions about their mutant: Is the mutant allele dominant or recessive to the wild-type allele? What chromosome does it map to? Between what other genes does it map? Is it an allele of a known gene? In summary, C. elegans is ideally suited for an undergraduate laboratory course. With appropriate training, undergraduate students can quickly learn to manipulate nematodes for use in genetic and/or molecular experiments. Furthermore, the flexibility of the 3-, 4-, or 7-day life cycle makes C. elegans particularly appealing for institutions on the quarter system. Access to teaching materials for both courses will be available at the teaching poster session and on my web page (www-acc.scu.edu/~lmiller/homepage.html).
-
[
International Worm Meeting,
2013]
At large public universities, it is often difficult to provide students with the optimum breadth and depth of laboratory experiences. To cope with the realities of team teaching, I have developed C. elegans based modules for three different lab courses: Introductory Biology (taught to 500 freshmen per semester), Genetics Lab (~50 upperclassmen per semester), and Cell Biology Lab (16 upperclassmen per semester). These modules provide a way to use C. elegans to introduce students to multiple subjects and techniques at different levels of complexity. Introductory Biology: The most basic module takes two meetings in the middle of the first term of a joint lecture-lab course; students have studied basic genetics and are studying cells. In week 1, students receive plates of N2,
dpy-5, and
unc-52. The students observe and record body bends per minute using a dissecting scope and learn about mutant phenotypes. In week 2, they analyze their data using Excel and use a shared fluorescent microscope to observe and photograph GFP+ worms. Genetics Lab: The module in this team-taught lab (which also uses Drosophila, bacteria, and yeast) introduces students to RNAi and gives them more experience with fluorescent microscopy. In week 1, students receive a regular or RNAi-hypersensitive GFP+ strain and plates with OP50 or
dpy-5 RNAi bacteria. In week 2, the students use upright (fluorescent) microscopes to observe the morphology of their treated worms vs.
dpy-5 mutants. The students take photographs and quantify worm length using ImageJ and Excel. Cell Biology Lab: "My" lab makes extensive uses of transgenic nematodes. Each student receives a different GFP-translational fusion strain near the beginning of the course. As an introduction to bioinformatics, the students use WormBase to gather information on their strains and GFP labeled proteins. The students use their strains to run protein gels and Westerns. They use fluorescent teaching scopes with digital cameras to observe and photograph their strains. They perform indirect immunofluorescence with an anti-GFP antibody, a second antibody, and phalloidin. The students are given supervised time at the confocal microscope to take images of their strain. The final class presentations allow the students to discuss the cell biology of 'their' GFP-proteins with each other.
-
[
International C. elegans Meeting,
1999]
This NSF-sponsored project integrates research and education by developing an intensive new laboratory course in which undergraduate students participate in a supervised research project. This research project is directly related to ongoing research in the laboratory of the principal investigator. The course is called "Project Lab" and is taught for one quarter each year with a class size of 7-16 students. Each year, the course focuses on research activities appropriate for that particular stage of the research project. The addition of this course to our biology curriculum enhances the science education of undergraduate students at Santa Clara University by providing hands-on laboratory experience and exposure to real research problems. It also enhances the research program of the principal investigator by training and motivating students for research. Specifically, students isolate and study mutations in genes that are required for cell fate determination in the process of vulval development in the nematode, C. elegans . The identification and analysis of these genes will set the stage for future experiments in cell fate determination. C. elegans is ideally suited for an undergraduate laboratory course. With appropriate training, undergraduate students can quickly learn to manipulate nematodes for use in genetic and/or molecular experiments. The first two Project Lab classes have made considerable progress on two different projects. For the first project, Project Lab students isolated and characterized over 30 new mutations that represents genes involved in the proper choice of cell fate during C. elegans vulval development. For the second project, several Project Lab students have been involved in a structure/function analysis of a transcription factor required for proper patterning of cell fate in C. elegans vulval development. There is a manuscript in preparation for this project, which will be submitted for publication within the next year. Three Project Lab students will be authors on this paper. I have also successfully used C. elegans in my upper division genetics laboratory course. This is a 10-week course in which students use nematodes to study inheritance patterns and map genes (using STS mapping).
-
[
International Worm Meeting,
2005]
Last summer, I designed and taught an Advanced Genetics Laboratory class that complements my own research project on the C. elegans defecation cycle. Students were introduced to genetic principles, classical and modern genetic mapping techniques, experimental analysis and Wormbase by characterizing and mapping defecation cycle mutants. For ten weeks, senior students attended two three-hour class periods and worked independently during open access laboratory periods. Disrupting the defecation cycle results in a distended intestinal lumen or a constipated (Con) worm. Since the Con phenotype is rather challenging to identify, the students analyzed Unc mutants for the first portion of the class. The first weeks were spent familiarizing students with worm handling and normal appearance. To facilitate this process, the students made mating plates and were tested on their ability to identify different sexes, stages, and mutant phenotypes. Next, each student was given a known Unc to map to a chromosome and to an approximate map position using classical chromosomal mapping techniques. Using Wormbase, students compared their Unc with Uncs in a similar genetic location and attempted to identify which Unc they had been given. For analysis of the Con mutant, the students were put into groups of three. Each member characterized the Con mutant's defecation cycle defect and completed two complementation tests with known Con mutants. As a group they pooled recombinants for bulk and fine SNIP-SNP mapping experiments using a genome wide Dra I SNIP-SNP primer set (designed by M. Hammarland and M. W. Davis). Grading was based on a quiz, two assignments - mating plate, defecation cycle characterization, two individual lab reports - mapping of the known Unc, and Con complementation testing and two group lab reports - bulk and fine SNP mapping of the Con. Students quickly learned worm techniques and were highly motivated to pursue "real" experiments. The students were able to make significant scientific contributions: a new allele of a known Con was identified by complementation testing and a novel Con was mapped to the middle of the LGX. Other aspects of the class could be improved such as experimental analysis and grading criteria. I welcome suggestions for improvement.
-
[
Mid-west Worm Meeting,
2002]
Faced with task of designing a semester's worth of comparative physiology laboratory exercises in my first semester as an assistant professor, I turned to my research and found what I had always suspected: worms are as wonderful in the teaching laboratory as they are in the research laboratory. As a supplement to several traditional physiology laboratory exercises, this past Spring semester, I required my physiology students to design and implement physiology research projects involving C. elegans. Although I tended to guide the student's projects as little as possible, the level of instructor involvement can be quite variable. By requiring students to write a research proposal prior to initiating their experiments, they can be directed towards realistic projects likely to succeed. The only stipulations were that 1. the research investigate some area of physiology discussed in lecture and 2. they not reproduce published data or experiments. At the first laboratory meeting, I introduced C. elegans as a research organism and asked them to consider their favorite topic from lecture. From this topic, I guided them towards the relevant C. elegansliterature and ultimately to a research plan. Prior to spring break, the students had solidified a research plan that included a hypothesis, a detailed experimental design, and identification of the relevant controls. This way, during the break, I was able to gather the required reagents and supplies. As the groups begin their experimentation, it is important to regularly check their progress in case experimental design troubleshooting is required. Ultimately, the students were responsible for initiating the experiments, limited troubleshooting, data collection, and assembly of the data into a research paper. I required that the research papers adhere to a specific journal format, typically one that is readily available for examples such as Current Biology. In this way, they experience the many steps in the process of converting great research ideas into meaningful publications. In the future, I plan to incorporate peer review into the process by having their papers reviewed by their student peers and other biology faculty prior to completion of the project. At the end of the semester, I required the students to present their hypothesis, experimental design, data, and conclusions to their colleagues in order to gain experience in oral research presentations and expose all the students to the various research projects. At the meeting, I will present the data collected by four of my undergraduate research teams who investigated many areas of physiology including aging, chemosensory adaptation, neurotransmitters, and serotonin modulated behavior.
-
[
International Worm Meeting,
2003]
A three-week series of C. elegans-based laboratories has been implemented in an introductory college genetics course (JS) that is based upon those used successfully in a high school program (KMM) and others (Jennifer Vowels, Debbie Birnby, Wendy Schackwitz, Jim Thomas). These labs are performed near the end of the semester, after students have been introduced to basic molecular biology concepts and at the same time that the lecture discusses the use of genetics to study behavior. The first session is a computer-based laboratory to introduce students to C. elegans and the vast array of information available at the C. elegans WWW server, Wormbase, and NCBI. Students are guided through questions and prompts to learn how to find different types of information such as: the literature published regarding chemotaxis in C. elegans, the names of researchers working on this problem, and the proteins that have been implicated in regulating chemotaxis. The second lab introduces students to the worm via three activities: 1) distinguishing males, adult hermaphrodites, and L4 worms; 2) identifying basic anatomical parts of the worm; and 3) sorting unc, dpy, and rol worms from a plate containing all three mutants. The first and third activity give students the opportunity to learn how to pick worms correctly, a skill they will need in the third lab. At the end of the second lab, students are assigned three unknowns (that they will identify in the third lab via chemotaxis assays) and pick worms to fresh plates in preparation for lab 3. The unknowns include two osm mutants, 1 che mutant, 1 che mutant marked with dpy, and wild-type. The third lab includes an attractance assay (based on Bargmann et al., 1993) and an avoidance assay, each performed with the three unknowns and wild-type controls. Students calculate avoidance and attractance indexes and perform statistical analyses to determine the identity of their unknowns. This series of laboratories could easily be expanded to incorporate student-directed projects, investigating other possible effectors of C. elegans behavior. Bargmann, C.I., Hartweig, E., and Horvitz, H.R. 1993. Odorant-selective genes and neurons mediate olfaction in C. elegans. Cell 74: 515-527.
-
[
International Worm Meeting,
2003]
Molecular Biology at Lawrence enrolls 35-45 undergraduates annually and includes three lectures and one three-hour lab/discussion section per week. Students have access to the lab and are expected to complete lab work on their own.Module 1, PCR: In previous years, students used PCR to identify deletions in genes
unc-93 or
unc-54. Students were given primer sets known to identify deletions in particular strains as well as plates of wild type and mutant animals. Students did a crude DNA extraction, set up the reactions, and analyzed PCR products using an acrylamide gel. This year Ive included site-directed mutagenesis. Student lab groups were given a computer file containing the
unc-54 gene sequence and told to design a particular kind of mutagenic primer and an upstream primer covering a restriction site. Students used computer programs to test the binding specificity of their primers and to predict the product size. Each lab section settled on one primer design. Over the course of two weeks, students prepared PCRs, digested PCR products as well as a plasmid containing
unc-54 with the chosen restriction enzymes, and purified the products from a low-melt agarose gel. The module concluded with production of ligation reactions which were handed to a research student for further analysis. If a submitted NSF grant proposal is funded, this laboratory exercise will continue for a third week next year when students will analyze their putative mutant constructs using an automated DNA sequencer.Module 2, Reporter Genes: In this qualitative, easy lab students learn the value of reporter gene constructs for analysis of gene expression patterns. It is a good lab to schedule at the end of a spring course when seniors dont want to work hard. Students are provided with two different mixed populations of worms, identified by number, each containing an lac-Z::promter fusion as an integrated transgenic array. Students permeabilize the worms and stain for B-galactosidase activity. Students carefully observe staining of different tissues in different ages of worms and draw examples of their observations before being told which promoter fusion they had. Students then use available databases to confirm their observations. Promoters used successfully thus far include
myo-2,
unc-54, and
pes-10 (the generous gift of Bill Morgan, College of Wooster). The use of lac-Z rather than GFP allows simultaneous observations by many students using simple dissecting microscopes. I would be happy to receive other strains containing other lac-Z fusions!
-
[
International Worm Meeting,
2005]
We have developed a C. elegans chemotaxis laboratory for use in an undergraduate introductory molecular biology course MOL101: From DNA to human complexity. This lecture and laboratory course is designed for non-science majors and fulfills a course distribution requirement for humanities majors at Princeton University.The chemotaxis lab is performed during the week when the lectures focus on behavioral genetics. It is designed to allow the students to be able to measure a behavioral response in C. elegans. Additionally, there is a student directed research component of the lab that allows students to design their own behavioral assays.In the first part of the lab, students work in pairs and set up a standard C. elegans chemotaxis assay. Briefly, about 100 animals are placed in the center of a 10 cm chemotaxis plate set up with isoamyl alcohol on one side and ethanol on the other side. Students calculate the chemotactic index after an hour has elapsed. During this time the students then design their own behavioral assay. They are provided with a number of different variables to choose from, including different attractants and repellents, frozen 2cm vials of acetic acid to change the temperature of the plates, and a sensory mutant strain. Students review their experimental design with a lab instructor prior to carrying out their designed experiment. In the following laboratory meeting, lab partners present their experimental design, results and conclusions in a 5 minute oral presentation.We have used this lab for two years and it is always successful. Aside from learning how to design a controlled experiment and how to correctly interpret results, students in the introductory class enjoy the chance to design their own experiments. Surprisingly, even with 12 groups in a section, most experiments are completely different from one another. An added benefit to this lab is that students do not need to learn how to manipulate or transfer individual worms, but instead can wash plates to collect worms and transfer them in bulk for the assay.
-
[
International Worm Meeting,
2003]
Faced with the task of designing a semester of physiology laboratory exercises in my first semester as an assistant professor, I turned to my research and found what I had always suspected: worms are as wonderful in the teaching laboratory as they are in the research laboratory. As a supplement to several traditional physiology laboratory exercises, for the past two Spring semesters I have required my comparative physiology students to design and implement physiology research projects involving C. elegans. Although I tend to guide the student's projects as little as possible, the level of instructor involvement can be quite variable. By requiring students to write a research proposal prior to initiating their experiments, they can be directed towards realistic projects likely to succeed. The only stipulations were that 1. the research investigate some area of physiology and 2. they not reproduce published data or experiments. At the first laboratory meeting, I introduce C. elegans as a research organism and ask them to consider their favorite area of physiology. From this interest, I guided them towards the relevant C. elegans literature and ultimately to a research plan. Prior to spring break, the students had solidified a research plan that included a hypothesis, a detailed experimental design, and identification of the relevant controls. This way, during the break, I am able to gather the required reagents, strains, and supplies. As the groups begin their experimentation, it is important to regularly check their progress in the event that experimental design troubleshooting is required. Ultimately, the students are responsible for initiating the experiments, limited troubleshooting, data collection, and assembly of the data into a research paper. I required that the research papers adhere to a specific journal format, typically one that is readily available for examples such as Current Biology. In this way, they experience the many steps in the process of converting great research ideas into meaningful publications. In the future, I plan to incorporate peer review into the process by having their papers reviewed by their student peers and other biology faculty prior to completion of the project. At the end of the semester, I also require the students to present their hypothesis, experimental design, data, and conclusions to their colleagues in order to gain experience in oral research presentations and expose all the students to the various research projects. At the meeting, I will present the data collected by four of my undergraduate research teams who investigated many areas of physiology including chemosensory adaptation, learning, and Pseudomonas pathogenecity.
-
[
International C. elegans Meeting,
2001]
The Rutgers University Department of Molecular Biology and Biochemistry has developed an undergraduate lab course that fully integrates C. elegans into a discovery-based research project that spans a full semester. "Introduction to Molecular Biology and Biochemistry Research" is taught by faculty members of the Department of MBB. The goal of this course is to allow students to investigate actual "unknown genes" while learning the basics of scientific research in the field of molecular biology. The lab is designed so that students, working in groups of two, randomly pick their own clones from a C. elegans embryonic cDNA library (gift of F. Piano) and analyze these clones over the course of the semester. In individual lab experiments the students are required to isolate the plasmid DNA, perform restriction enzyme digests, perform gel electrophoresis and prepare the DNA for sequencing. Students are then able to perform computer-based bioinformatic analysis of the obtained sequences. Finally, students are able to directly asses the developmental function of their gene by performing RNAi feeding experiments with C. elegans . The specifics of how the laboratory experiments are carried out will be presented. Briefly, the cDNA library was constructed in the TriplEx vector. This vector contains two T& promoter sites flanking the insert to allow expression of dsRNA when transformed into an appropriate bacterial strain. Students are therefore able to interpret the role of their gene in embryonic development by examining worms grown on bacteria that produce dsRNA of their specific genes. This is accomplished through use of the
lin-2 strain that normally produces a bag of worms phenotype. If the dsRNA disrupts embryonic development, then a bag of eggs phenotype will be observed. This version of the Introduction to Molecular Biology and Biochemistry Research laboratory course has been completed twice. We have found on average that about one third of the clones tested by the students exhibit an embryonic lethal phenotype.