Today ’ s Goals
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Today’s Goals
Describe the advantages of C. elegans as a model organism
Discuss the life cycle of the nematodeSafely and effectively culture a
population of C. elegans and transfer worms to new plates for experimental studies
The model organism: Caenorhabditis elegans
Electron micrograph of a C. elegans hermaphrodite
Small = about 1mm in sizeLives in the soilNematodeEats bacteriaCan be male or hermaphroditic
Self-fertilizing!
Caenorhabditis elegans
Profile
Soil nematode, EukaryoteGenome size: 100 MbNumber of chromosomes: 6Generation time: about 2 daysFemale reproductive capacity: 250 to 1000 progeny
Special characteristicsStrains Can Be FrozenEasy to growHermaphroditeKnown cell lineage pattern for all 959 somatic cellsOnly 302 neuronsTransparent bodyCan be characterized geneticallyAbout 70% of Human Genes have related genes in C. elegans
Lifecycle of a worm
(from www.wormatlas.org)
Anatomy of a worm
(from www.wormatlas.org)
C. elegans cell division can be studied in the transparent egg
C. elegans cell lineage is known
Fire & Mello - Won Nobel prize for discovery of RNAi
C. elegans is amenable to many forms of RNAi treament
Feeding worms bacteria that express dsRNAs or soaking worms in dsRNA sufficient to induce silencing (Gene 263:103, 2001; Science 282:430, 1998)
C. elegans can also be used for Behavioral studies
Will respond when tapped -Different mutants might respond differently
Display chemotaxisMove toward certain chemicals
Studying Neurobiology Very few neurons Easy to map out what neurons target each part of the body, test
genetics of neurobiology
Studying Embryonic Development Cell biology, Apoptosis . . . And more !!! C. Elegans in space!!!
Nobel Prize winners who did their research with C. elegansSydney BrennerJohn SulstonBob HorvitzAndrew FireCraig Mello
http://park.itc.utokyo.ac.jp/mgrl/IINO_lab/Eng_figures/Fig4.JPEG&imgrefurl=http://park.itc.utokyo.ac.jp/mgrl/IINO_lab/IINO_lab.html&h=491&w=333&sz=29&hl=en&start=23&um=1&tbnid=6D_KWwsfwR_pZM:&tbnh=130&tbnw=88&prev=/images%3Fq%3Dc.%2Belegans%2Bchemotaxis%26start%3D20%26ndsp%3D20%26um%3D1%26hl%3Den%26client%3Dsafari%26rls%3Den%26sa%3DN
How will we be working with C. elegans?We will be following along with lab
protocols from www.silencinggenomes.org Learn to culture and examine nematodes Learn techniques to count and transfer
worms to separate dishes Perform RNAi, using a kit Design experiments of our own with
techniques we read about in the primary literature
We can order C. elegans MutantsUniversity of MinnesotaCaenorhabditis genetics center7$ per strain
http://www.cbs.umn.edu/CGC/
NGM Plates and OP50 broth
You will pour NGM-lite plates Pour 1 bottle per pair of students
Using Sterile Technique Nematode Growth Medium Label Plates NGM, Date, Initials
When solid, invert, put on lab bench by incubators Then – take 5 microliters of OP50 bacteria,
innoculate a tube of LB broth (on lab bench) Label with your initials, and OP50, put in 37 Degree
incubator Next time - we’ll seed plates with bacteria for
culture of C. elegans