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Aldicarb-induced Paralysis Assay to Determine Defects in Synaptic Transmission in Caenorhabditis elegans

Featured protocol,  Authors: Kelly H. Oh
Kelly H. OhAffiliation: Department of Cell Biology and Anatomy, Chicago Medical School, Rosalind Franklin University of Medicine and Science, North Chicago, USA
Bio-protocol author page: a4871
 and Hongkyun Kim
Hongkyun KimAffiliation: Department of Cell Biology and Anatomy, Chicago Medical School, Rosalind Franklin University of Medicine and Science, North Chicago, USA
For correspondence: Hongkyun.kim@rosalindfranklin.edu
Bio-protocol author page: a4870
date: 7/20/2017, 18 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2400.

Brief version appeared in Elife, Feb 2017
Aldicarb treatment causes an accumulation of acetylcholine in the synaptic cleft of the neuromuscular junction, resulting in sustained muscle activation and eventually paralysis. Aldicarb-induced paralysis assay is an easy and fast method to determine whether synaptic transmission of a C. elegans mutant of interest is altered. This assay is based on the correlation of the rate of neurotransmitter release with the rate of paralysis. In this protocol, we describe a method for simultaneously assessing the aldicarb sensitivity of animals with different genotypes.

Oxidative Stress Assays (arsenite and tBHP) in Caenorhabditis elegans

Featured protocol,  Authors: Collin Yvès Ewald
Collin Yvès EwaldAffiliation 1: Department of Health Sciences and Technology, Eidgenössische Technische Hochschule (ETH) Zürich, Schwerzenbach-Zürich, Switzerland
Affiliation 2: Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA
Affiliation 3: Harvard Stem Cell Institute, Harvard University, Boston, Massachusetts, USA
Affiliation 4: Joslin Diabetes Center, Research Division, Boston, Massachusetts, USA
For correspondence: collin-ewald@ethz.ch
Bio-protocol author page: a4749
John M. Hourihan
John M. HourihanAffiliation 1: Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA
Affiliation 2: Harvard Stem Cell Institute, Harvard University, Boston, Massachusetts, USA
Affiliation 3: Joslin Diabetes Center, Research Division, Boston, Massachusetts, USA
Bio-protocol author page: a4750
 and T. Keith Blackwell
T. Keith BlackwellAffiliation 1: Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA
Affiliation 2: Harvard Stem Cell Institute, Harvard University, Boston, Massachusetts, USA
Affiliation 3: Joslin Diabetes Center, Research Division, Boston, Massachusetts, USA
Bio-protocol author page: a4751
date: 7/5/2017, 254 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2365.

Brief version appeared in Elife, Jan 2017
Cells and organisms face constant exposure to reactive oxygen species (ROS), either from the environment or as a by-product from internal metabolic processes. To prevent cellular damage from ROS, cells have evolved detoxification mechanisms. The activation of these detoxification mechanisms and their downstream responses represent an overlapping defense response that can be tailored to different sources of ROS to adequately adapt and protect cells. In this protocol, we describe how to measure the sensitivity to oxidative stress from two different sources, arsenite and tBHP, using the nematode C. elegans.

Aldicarb-induced Paralysis Assay to Determine Defects in Synaptic Transmission in Caenorhabditis elegans

Authors: Kelly H. Oh
Kelly H. OhAffiliation: Department of Cell Biology and Anatomy, Chicago Medical School, Rosalind Franklin University of Medicine and Science, North Chicago, USA
Bio-protocol author page: a4871
 and Hongkyun Kim
Hongkyun KimAffiliation: Department of Cell Biology and Anatomy, Chicago Medical School, Rosalind Franklin University of Medicine and Science, North Chicago, USA
For correspondence: Hongkyun.kim@rosalindfranklin.edu
Bio-protocol author page: a4870
date: 7/20/2017, 18 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2400.

[Abstract] Aldicarb treatment causes an accumulation of acetylcholine in the synaptic cleft of the neuromuscular junction, resulting in sustained muscle activation and eventually paralysis. Aldicarb-induced paralysis assay is an easy and fast method to determine whether synaptic transmission of a C. elegans mutant ...

Oxidative Stress Assays (arsenite and tBHP) in Caenorhabditis elegans

Authors: Collin Yvès Ewald
Collin Yvès EwaldAffiliation 1: Department of Health Sciences and Technology, Eidgenössische Technische Hochschule (ETH) Zürich, Schwerzenbach-Zürich, Switzerland
Affiliation 2: Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA
Affiliation 3: Harvard Stem Cell Institute, Harvard University, Boston, Massachusetts, USA
Affiliation 4: Joslin Diabetes Center, Research Division, Boston, Massachusetts, USA
For correspondence: collin-ewald@ethz.ch
Bio-protocol author page: a4749
John M. Hourihan
John M. HourihanAffiliation 1: Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA
Affiliation 2: Harvard Stem Cell Institute, Harvard University, Boston, Massachusetts, USA
Affiliation 3: Joslin Diabetes Center, Research Division, Boston, Massachusetts, USA
Bio-protocol author page: a4750
 and T. Keith Blackwell
T. Keith BlackwellAffiliation 1: Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA
Affiliation 2: Harvard Stem Cell Institute, Harvard University, Boston, Massachusetts, USA
Affiliation 3: Joslin Diabetes Center, Research Division, Boston, Massachusetts, USA
Bio-protocol author page: a4751
date: 7/5/2017, 254 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2365.

[Abstract] Cells and organisms face constant exposure to reactive oxygen species (ROS), either from the environment or as a by-product from internal metabolic processes. To prevent cellular damage from ROS, cells have evolved detoxification mechanisms. The activation of these detoxification mechanisms and their ...

Culturing Bacteria from Caenorhabditis elegans Gut to Assess Colonization Proficiency

Authors: Facundo Rodriguez Ayala
Facundo Rodriguez AyalaAffiliation: Departamento de Microbiología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, CONICET, Rosario, Argentina
For correspondence: facundoayala.foncyt@gmail.com
Bio-protocol author page: a4690
Sebastián Cogliati
Sebastián CogliatiAffiliation: Departamento de Microbiología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, CONICET, Rosario, Argentina
Bio-protocol author page: a4691
Carlos Bauman
Carlos BaumanAffiliation: Departamento de Microbiología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, CONICET, Rosario, Argentina
Bio-protocol author page: a4692
Cecilia Leñini
Cecilia LeñiniAffiliation: Departamento de Microbiología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, CONICET, Rosario, Argentina
Bio-protocol author page: a4693
Marco Bartolini
Marco BartoliniAffiliation: Departamento de Microbiología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, CONICET, Rosario, Argentina
Bio-protocol author page: a4694
Juan Manuel Villalba
Juan Manuel VillalbaAffiliation: Departamento de Microbiología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, CONICET, Rosario, Argentina
Bio-protocol author page: a4695
Federico Argañaraz
Federico ArgañarazAffiliation: Departamento de Microbiología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, CONICET, Rosario, Argentina
Bio-protocol author page: a4696
 and Roberto Grau
Roberto GrauAffiliation: Departamento de Microbiología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, CONICET, Rosario, Argentina
Bio-protocol author page: a4697
date: 6/20/2017, 305 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2345.

[Abstract] Determining an accurate count of intestinal bacteria from Caenorhabditis elegans is one critical way to assess colonization proficiency by a given bacteria. This can be accomplished by culturing appropriate dilutions of worm gut bacteria on selective or differential agarized media. Because of the high ...

Single-molecule RNA Fluorescence in situ Hybridization (smFISH) in Caenorhabditis elegans

Authors: ChangHwan Lee
ChangHwan LeeAffiliation 1: Howard Hughes Medical Institute, University of Wisconsin-Madison, Madison, Wisconsin, 53706, USA
Affiliation 2: Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin, USA
Bio-protocol author page: a4726
Hannah S. Seidel
Hannah S. SeidelAffiliation 1: Howard Hughes Medical Institute, University of Wisconsin-Madison, Madison, Wisconsin, 53706, USA
Affiliation 2: Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin, USA
Present address: Department of Biology, Eastern Michigan University, Ypsilanti, Michigan, USA
Bio-protocol author page: a4727
Tina R. Lynch
Tina R. LynchAffiliation: Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin, USA
Bio-protocol author page: a4728
Erika B. Sorensen
Erika B. SorensenAffiliation 1: Howard Hughes Medical Institute, University of Wisconsin-Madison, Madison, Wisconsin, 53706, USA
Affiliation 2: Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin, USA
Present address: Department of Biology, Wabash College, Crawfordsville, Indiana, USA
Bio-protocol author page: a4729
Sarah L. Crittenden
Sarah L. CrittendenAffiliation: Howard Hughes Medical Institute, University of Wisconsin-Madison, Madison, Wisconsin, 53706, USA
Bio-protocol author page: a4730
 and Judith Kimble
Judith KimbleAffiliation 1: Howard Hughes Medical Institute, University of Wisconsin-Madison, Madison, Wisconsin, 53706, USA
Affiliation 2: Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin, USA
For correspondence: jekimble@wisc.edu
Bio-protocol author page: a4731
date: 6/20/2017, 293 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2357.

[Abstract] Single-molecule RNA fluorescence in situ hybridization (smFISH) is a technique to visualize individual RNA molecules using multiple fluorescently-labeled oligonucleotide probes specific to the target RNA (Raj et al., 2008; Lee et al., 2016a). We adapted this technique to visualize RNAs in the C. elegans ...

Heavy Metal Stress Assay of Caenorhabditis elegans

Authors: Strahil Iv. Pastuhov
Strahil Iv. PastuhovAffiliation: Division of Biological Science, Graduate School of Science, Nagoya University, Chikusa-ku, Nagoya 464-8602, Japan
Bio-protocol author page: a4596
Tatsuhiro Shimizu
Tatsuhiro ShimizuAffiliation: Division of Biological Science, Graduate School of Science, Nagoya University, Chikusa-ku, Nagoya 464-8602, Japan
Bio-protocol author page: a4597
 and Naoki Hisamoto
Naoki HisamotoAffiliation: Division of Biological Science, Graduate School of Science, Nagoya University, Chikusa-ku, Nagoya 464-8602, Japan
For correspondence: i45556a@cc.nagoya-u.ac.jp
Bio-protocol author page: a4598
date: 6/5/2017, 455 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2312.

[Abstract] Organisms have developed many protective systems to reduce the toxicity from heavy metals. The nematode Caenorhabditis elegans has been widely used to determine the protective mechanisms against heavy metals. Responses against heavy metals can be monitored by expression of reporter genes, while sensitivity ...

Imaging the Pharynx to Measure the Uptake of Doxorubicin in Caenorhabditis elegans

Authors: Sivathevy Amirthagunabalasingam
Sivathevy AmirthagunabalasingamAffiliation: Maisonneuve-Rosemont Hospital Research Center, and the Université de Montréal, Faculty of Medicine, Department of Medicine, Montréal, Canada
Bio-protocol author page: a4534
Arturo Papaluca
Arturo PapalucaAffiliation: , Lady Davis Institute, McGill University, Montréal, Canada
Bio-protocol author page: a4535
Taramatti Harihar
Taramatti HariharAffiliation: Maisonneuve-Rosemont Hospital Research Center, and the Université de Montréal, Faculty of Medicine, Department of Medicine, Montréal, Canada
Bio-protocol author page: a4536
 and Dindial Ramotar
Dindial RamotarAffiliation: Maisonneuve-Rosemont Hospital Research Center, and the Université de Montréal, Faculty of Medicine, Department of Medicine, Montréal, Canada
For correspondence: dindial.ramotar@umontreal.ca
Bio-protocol author page: a4537
date: 5/20/2017, 375 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2291.

[Abstract] Caenorhabditis elegans offers an array of advantages to investigate the roles of uptake transporters. Herein, an epifluorescent microscopy approach was developed to monitor the uptake of the autofluorescent anticancer drug, doxorubicin, into the pharynx of C. elegans by organic cation transporters....

Nuclei Isolation from Nematode Ascaris

Authors: Yuanyuan Kang
Yuanyuan KangAffiliation: Department of Biochemistry and Molecular Genetics, University of Colorado School of Medicine, Aurora, USA
Bio-protocol author page: a4463
Jianbin Wang
Jianbin WangAffiliation: Department of Biochemistry and Molecular Genetics, University of Colorado School of Medicine, Aurora, USA
Bio-protocol author page: a4464
 and Richard E. Davis
Richard E. DavisAffiliation: Department of Biochemistry and Molecular Genetics, University of Colorado School of Medicine, Aurora, USA
For correspondence: richard.davis@ucdenver.edu
Bio-protocol author page: a4465
date: 5/5/2017, 477 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2262.

[Abstract] Preparing nuclei is necessary in a variety of experimental paradigms to study nuclear processes. In this protocol, we describe a method for rapid preparation of large number of relatively pure nuclei from Ascaris embryos or tissues that are ready to be used for further experiments such as chromatin ...

Electroshock Induced Seizures in Adult C. elegans

Authors: Monica G Risley
Monica G RisleyAffiliation 1: Department of Biological Sciences, Florida Atlantic University, Boca Raton, USA
Affiliation 2: International Max Planck Research School (IMPRS) for Brain and Behavior, Boca Raton, USA
Bio-protocol author page: a4483
Stephanie P Kelly
Stephanie P KellyAffiliation: Department of Biological Sciences, Florida Atlantic University, Boca Raton, United States
Bio-protocol author page: a4484
 and Ken Dawson-Scully
Ken Dawson-ScullyAffiliation: Department of Biological Sciences, Florida Atlantic University, Boca Raton, USA
For correspondence: ken.dawson-scully@fau.edu
Bio-protocol author page: a4485
date: 5/5/2017, 395 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2270.

[Abstract] The nematode Caenorhabditis elegans is a useful model organism for dissecting molecular mechanisms of neurological diseases. While hermaphrodite C. elegans contain only 302 neurons, the conserved homologous neurotransmitters, simpler neuronal circuitry, and fully mapped connectome make it an appealing ...

In vivo Mitophagy Monitoring in Caenorhabditis elegans to Determine Mitochondrial Homeostasis

Authors: Konstantinos Palikaras
Konstantinos PalikarasAffiliation: Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology Hellas, Crete, Greece
Bio-protocol author page: a3832
 and Nektarios Tavernarakis
Nektarios TavernarakisAffiliation 1: Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology Hellas, Crete, Greece
Affiliation 2: Department of Basic Sciences, Faculty of Medicine, University of Crete, Crete, Greece
For correspondence: tavernarakis@imbb.forth.gr
Bio-protocol author page: a3833
date: 4/5/2017, 573 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2215.

[Abstract] Perturbation of mitochondrial function is a major hallmark of several pathological conditions and ageing, underlining the essential role of fine-tuned mitochondrial activity (Lopez-Otin et al., 2013). Mitochondrial selective autophagy, known as mitophagy, mediates the removal of dysfunctional and/or ...

In vivo Live Imaging of Calcium Waves and Other Cellular Processes during Fertilization in Caenorhabditis elegans

Authors: Jun Takayama
Jun TakayamaAffiliation: RIKEN Quantitative Biology Center, Laboratory for Developmental Dynamics, Kobe, Japan
Bio-protocol author page: a4310
Masashi Fujita
Masashi FujitaAffiliation: RIKEN Quantitative Biology Center, Laboratory for Developmental Dynamics, Kobe, Japan
Bio-protocol author page: a4311
 and Shuichi Onami
Shuichi OnamiAffiliation: RIKEN Quantitative Biology Center, Laboratory for Developmental Dynamics, Kobe, Japan
For correspondence: sonami@riken.jp
Bio-protocol author page: a4312
date: 4/5/2017, 766 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2205.

[Abstract] Fertilization calcium waves are a conserved trigger for animal development; however, genetic analysis of these waves has been limited due to the difficulty of imaging in vivo fertilization. Here we describe a protocol to image calcium dynamics during in vivo fertilization in the genetic animal model ...
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[Bio101] Common Worm Media and Buffers

Author: Fanglian He date: 4/5/2011, 24588 views, 11 Q&A
DOI: https://doi.org/10.21769/BioProtoc.55.

[Abstract] Here are recipes of some media and solutions often used in C. elegans research....

[Bio101] Lifespan Assay

Author: Fanglian He date: 4/5/2011, 15132 views, 3 Q&A
DOI: https://doi.org/10.21769/BioProtoc.57.

[Abstract] This assay is used to address aging-related questions in worms....

[Bio101] Total RNA Extraction from C. elegans

Author: Fanglian He date: 3/20/2011, 15001 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.47.

[Abstract] This protocol describes total RNA extraction from worms with or without using commercial RNA extraction kits....

[Bio101] Making Males of C. elegans

Author: Fanglian He date: 4/20/2011, 14229 views, 2 Q&A
DOI: https://doi.org/10.21769/BioProtoc.58.

[Abstract] Getting males from a hermaphrodite population. This is a modified version of protocol originally written by Michael Koelle at Yale University....

[Bio101] RNA Interference (RNAi) by Bacterial Feeding

Author: Fanglian He date: 4/20/2011, 14166 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.59.

[Abstract] There are 3 ways to perform RNAi in worms: microinjection, soaking and feeding. In the feeding protocol, RNAi is induced by cultivating worms on bacteria expressing gene-specific dsRNA. dsRNA is expressed in E. coli and ingested by worms. This protocol describes the feeding protocol to induce RNAi....

[Bio101] Synchronization of Worm

Author: Fanglian He date: 4/5/2011, 13629 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.56.

[Abstract] Attaining a large synchronized population of worms is desirable for use in some assays in order to eliminate variation in results due to age differences. Two ways to get synchronized worms include egg preparation via bleaching and egg lay. The former in general yields more progeny than the latter, however, ...

PCR-RFLP Genotyping of Point Mutations in Caenorhabditis elegans

Author: Peichuan Zhang
Peichuan ZhangAffiliation: Department of Biochemistry and Biophysics, University of California, San Francisco, USA
For correspondence: peichuan.zhang@ucsf.edu
Bio-protocol author page: a11
date: 3/20/2012, 12886 views, 3 Q&A
DOI: https://doi.org/10.21769/BioProtoc.128.

[Abstract] This protocol describes the basic principle of PCR/restriction digest genotyping of point mutations in worms, based on the principle of Restriction Fragment Length Polymorphism (RFLP) analysis. This type of genotyping is, particularly, useful when phenotypic analysis of animals carrying point mutations ...

[Bio101] Oil Red O Staining of Fixed Worm

Author: Fanglian He date: 7/5/2012, 12496 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.230.

[Abstract] Oil red O staining is used to assess major fat stores in C. elegans. This protocol is adapted from the Ashrafi Lab at the University of California-San Francisco....

[Bio101] DAPI Nuclear Staining of Live Worm

Author: Fanglian He date: 6/5/2011, 10840 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.77.

[Abstract] Adapted from the Villenueve Lab at Stanford University. This is a very simple method using ethanol fixation, but works very well....

[Bio101] Single Worm PCR

Author: Fanglian He date: 4/20/2011, 10313 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.60.

[Abstract] This protocol is used for genotyping single worms....
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