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    Protocols in Current Issue
    Bioluminescence Monitoring of Neuronal Activity in Freely Moving Zebrafish Larvae
    The proof of concept for bioluminescence monitoring of neural activity in zebrafish with the genetically encoded calcium indicator GFP-aequorin has been previously described (Naumann et al., 2010) but challenges remain. First, ...
    Stereotaxic Adeno-associated Virus Injection and Cannula Implantation in the Dorsal Raphe Nucleus of Mice
    Authors:  Patrícia A. Correia, Sara Matias and Zachary F. Mainen, date: 09/20/2017, view: 31, Q&A: 0
    Optogenetic methods are now widespread in neuroscience research. Here we present a detailed surgical procedure to inject adeno-associated viruses and implant optic fiber cannulas in the dorsal raphe nucleus (DRN) of living mice. Combined with ...
    Preparation of Primary Cultures of Embryonic Rat Hippocampal and Cerebrocortical Neurons
    This protocol aims at standardizing the procedure to obtain primary cultures of hippocampal and cerebrocortical neurons for in vitro experiments. Cultures should be prepared from cells isolated during embryonic development when neuronal ...
    Bioluminescence Monitoring of Neuronal Activity in Freely Moving Zebrafish Larvae
    [Abstract] The proof of concept for bioluminescence monitoring of neural activity in zebrafish with the genetically encoded calcium indicator GFP-aequorin has been previously described (Naumann et al., 2010) but challenges remain. First, bioluminescence signals originating from a single muscle fiber can constitute a major pitfall. Second, ...
    Stereotaxic Adeno-associated Virus Injection and Cannula Implantation in the Dorsal Raphe Nucleus of Mice
    Authors:  Patrícia A. Correia, Sara Matias and Zachary F. Mainen, date: 09/20/2017, view: 31, Q&A: 0
    [Abstract] Optogenetic methods are now widespread in neuroscience research. Here we present a detailed surgical procedure to inject adeno-associated viruses and implant optic fiber cannulas in the dorsal raphe nucleus (DRN) of living mice. Combined with transgenic mouse lines, this protocol allows specific targeting of serotonin-producing neurons in the ...
    Preparation of Primary Cultures of Embryonic Rat Hippocampal and Cerebrocortical Neurons
    [Abstract] This protocol aims at standardizing the procedure to obtain primary cultures of hippocampal and cerebrocortical neurons for in vitro experiments. Cultures should be prepared from cells isolated during embryonic development when neuronal precursor cells are not yet fully differentiated. This helps increasing the quality and quantity of ...
    Arginine-rich Peptides Can Actively Mediate Liquid-liquid Phase Separation
    Authors:  Steven Boeynaems, Mathias De Decker, Peter Tompa and Ludo Van Den Bosch, date: 09/05/2017, view: 385, Q&A: 0
    [Abstract] Studying liquid-liquid phase separation (LLPS) of proteins provides key insights into the biogenesis of membraneless organelles and pathological protein aggregation in disease. We have established a protocol for inducing the phase separation of arginine-rich peptides, which allows for studying their molecular determinants and dynamics (Boeynaems ...
    Isolation of Rodent Brain Vessels
    Authors:  Cristina Carvalho and Paula I. Moreira, date: 09/05/2017, view: 159, Q&A: 0
    [Abstract] The prevalence of neurodegenerative diseases is increasing worldwide. Cerebrovascular disorders and/or conditions known to affect brain vasculature, such as diabetes, are well-known risk factors for neurodegenerative diseases. Thus, the evaluation of the brain vasculature is of great importance to better understand the mechanisms underlying brain ...
    Predator Odor-induced Freezing Test for Mice
    Author:  Shintaro Otsuka, date: 09/05/2017, view: 149, Q&A: 0
    [Abstract] The innate fear response is an emotional response that does not require any previously acquired conditioning. One of the standard methods to analyze the innate fear response is a 2,4,5-trimethylthiazoline (TMT)-induced freezing test. TMT is an odor originally isolated from anal secretion of the red fox. Acute TMT exposure has been shown to induce ...
    FM1-43 Photoconversion and Electron Microscopy Analysis at the Drosophila Neuromuscular Junction
    Authors:  Nadezhda S. Sabeva and Maria Bykhovskaia, date: 09/05/2017, view: 277, Q&A: 0
    [Abstract] We developed a protocol for photoconversion of endocytic marker FM1-43 followed by electron microscopy analysis of synaptic boutons at the Drosophila neuromuscular junction. This protocol allows detection of stained synaptic vesicle even when release rates are very low, such as during the spontaneous release mode. The preparations are ...
    An ex vivo Perifusion Method for Quantitative Determination of Neuropeptide Release from Mouse Hypothalamic Explants
    Authors:  Ophélia Le Thuc, Jacques Noël and Carole Rovère, date: 08/20/2017, view: 411, Q&A: 0
    [Abstract] The hypothalamus is a primary brain area which, in mammals, regulates several physiological functions that are all related to maintain general homeostasis, by linking the central nervous system (CNS) and the periphery. The hypothalamus itself can be considered an endocrine brain region of some sort as it hosts in its different nuclei several kinds ...
    A High-throughput Assay for mRNA Silencing in Primary Cortical Neurons in vitro with Oligonucleotide Therapeutics
    [Abstract] Primary neurons represent an ideal cellular system for the identification of therapeutic oligonucleotides for the treatment of neurodegenerative diseases. However, due to the sensitive nature of primary cells, the transfection of small interfering RNAs (siRNA) using classical methods is laborious and often shows low efficiency. Recent progress in ...
    Assessment of Thermal Pain Sensation in Rats and Mice Using the Hargreaves Test
    Authors:  Menghon Cheah, James W Fawcett and Melissa R Andrews, date: 08/20/2017, view: 334, Q&A: 0
    [Abstract] The Hargreaves test is specifically designed to assess thermal pain sensation in rodents such as rats and mice. This test has been used in experiments involving pain sensitization or recovery of thermal pain response following neural injury and regeneration. We present here a step-by-step protocol highlighted with important notes to guide ...