Protocols in Current Issue
    Staining the Germline in Live Caenorhabditis elegans: Overcoming Challenges by Applying a Fluorescent-dye Feeding Strategy
    Author:  K. Adam Bohnert, date: 11/05/2018, view: 172, Q&A: 0
    [Abstract] C. elegans provides a tractable model organism for studying germline cell biology. Microscopy experiments are relatively facile, as this worm is transparent and germline development can be observed in real-time using DIC microscopy and/or fluorescent transgenes. Despite these many tools, robust staining techniques for imaging germ cells ...
    Nuclear/Cytoplasmic Fractionation of Proteins from Caenorhabditis elegans
    Authors:  Alejandro Mata-Cabana, Olga Sin, Renée I. Seinstra and Ellen A. A. Nollen, date: 10/20/2018, view: 519, Q&A: 0
    [Abstract] C. elegans is widely used to investigate biological processes related to health and disease. To study protein localization, fluorescently-tagged proteins can be used in vivo or immunohistochemistry can be performed in whole worms. Here, we describe a technique to localize a protein of interest at a subcellular level in C. elegans ...
    Filter Retardation Assay for Detecting and Quantifying Polyglutamine Aggregates Using Caenorhabditis elegans Lysates
    Authors:  Olga Sin, Alejandro Mata-Cabana, Renée I. Seinstra and Ellen A. A. Nollen, date: 10/05/2018, view: 324, Q&A: 0
    [Abstract] Protein aggregation is a hallmark of several neurodegenerative diseases and is associated with impaired protein homeostasis. This imbalance is caused by the loss of the protein’s native conformation, which ultimately results in its aggregation or abnormal localization within the cell. Using a C. elegans model of polyglutamine diseases, we ...
    6-hydroxydopamine (6-OHDA) Oxidative Stress Assay for Observing Dopaminergic Neuron Loss in Caenorhabditis elegans
    Authors:  Sarah-Lena Offenburger and Anton Gartner, date: 09/20/2018, view: 316, Q&A: 0
    [Abstract] The nematode Caenorhabditis elegans is a powerful genetic model that can be used to investigate neuronal death. Research using C. elegans has been crucial to characterize cell death programmes that are conserved in mammals. Many neuronal signaling components, such as those mediating dopaminergic neurotransmission, are preserved ...
    A Behavioral Assay to Examine the Effects of Kavalactones on Caenorhabditis elegans Neuromuscular Excitability
    [Abstract] Kavalactones are a class of lactone compounds found in Kava, a traditional beverage from the South Pacific Islands that is derived from the root of Piper methysticum. When consumed, these compounds produce sedative and anxiolytic effects, suggesting their potent actions on the nervous system. Here, we provide a protocol to examine the ...
    Pentylenetetrazole (PTZ)-induced Convulsion Assay to Determine GABAergic Defects in Caenorhabditis elegans
    Authors:  Shruti Thapliyal and Kavita Babu, date: 09/05/2018, view: 356, Q&A: 0
    [Abstract] Pentylenetetrazole (PTZ) is a GABAA receptor antagonist and is used to monitor presynaptic defects in the release of the inhibitory neurotransmitter GABA. PTZ is a competitive inhibitor of GABA, and prevents binding of GABA on the GABAA receptors present on the surface of muscle. In the absence of GABA binding, the excitatory ...
    Assessment of Caenorhabditis elegans Competitive Fitness in the Presence of a Bacterial Parasite
    Authors:  McKenna. J. Penley and Levi T. Morran, date: 08/20/2018, view: 490, Q&A: 0
    [Abstract] Accurate measurements of an organism’s fitness are crucial for measuring evolutionary change. Methods of fitness measurement are most accurate when incorporating an individual’s survival and fecundity, as well as accounting for any ecological interactions or environmental effects experienced by the organism. Here, we describe a protocol for ...
    FRAP: A Powerful Method to Evaluate Membrane Fluidity in Caenorhabditis elegans
    Authors:  Ranjan Devkota and Marc Pilon, date: 07/05/2018, view: 937, Q&A: 0
    [Abstract] FRAP (Fluorescence Recovery After Photobleaching) is probably the most direct method to investigate the dynamics of molecules in living cells. Here, we describe FRAP to quantify membrane fluidity in C. elegans. Using FRAP, we have shown that cold, glucose and exogenous saturated fatty acids can decrease the fluidity of cellular membranes ...
    Measuring Spatiotemporal Dynamics of Odor Gradient for Small Animals by Gas Chromatography
    Authors:  Akiko Yamazoe-Umemoto, Yuishi Iwasaki and Koutarou D. Kimura, date: 04/05/2018, view: 1502, Q&A: 0
    [Abstract] Odor is the most fundamental chemical stimulus that delivers information regarding food, mating partners, enemies, and danger in the surrounding environment. Research on odor response in animals is widespread, although studies on experimental systems in which the gradient of odor concentration is quantitatively measured has been quite limited. ...
    Measurement of ROS in Caenorhabditis elegans Using a Reduced Form of Fluorescein
    Authors:  Shaarika Sarasija and Kenneth R. Norman, date: 04/05/2018, view: 1254, Q&A: 0
    [Abstract] Oxidative stress is implicated in the pathogenesis of various neurodegenerative diseases, including Alzheimer’s disease. Oxidative stress is a result of a disruption of the equilibrium between antioxidants and oxidants, in favor of oxidants. Since mitochondria are major sites of production and reduction of reactive oxygen species (ROS), ...

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