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    Protocols in Current Issue
    Culture and Nucleofection of Postnatal Day 7 Cortical and Cerebellar Mouse Astroglial Cells
    Authors:  Malek Chouchane and Marcos Romualdo Costa, date: 02/05/2018, view: 158, Q&A: 0
    Lineage reprogramming of astroglial cells isolated from different brain regions leads to the generation of different neuronal subtypes. This protocol describes the isolation and culture of neocortical and cerebellar astrocytes from postnatal mice. ...
    Determination of DNA Damage in the Retina Photoreceptors of Drosophila
    Authors:  Damulewicz Milena and Pyza Elzbieta, date: 02/05/2018, view: 79, Q&A: 0
    The retina is sensitive for light damages, because of direct light exposure, especially intense blue and UV light, which increase level of ROS and other toxic phototransduction products in photoreceptor cells. In our previous work (Damulewicz et al. ...
    Culture and Nucleofection of Postnatal Day 7 Cortical and Cerebellar Mouse Astroglial Cells
    Authors:  Malek Chouchane and Marcos Romualdo Costa, date: 02/05/2018, view: 158, Q&A: 0
    [Abstract] Lineage reprogramming of astroglial cells isolated from different brain regions leads to the generation of different neuronal subtypes. This protocol describes the isolation and culture of neocortical and cerebellar astrocytes from postnatal mice. We also present a comprehensive description of the main steps towards successful gene delivery in ...
    Determination of DNA Damage in the Retina Photoreceptors of Drosophila
    Authors:  Damulewicz Milena and Pyza Elzbieta, date: 02/05/2018, view: 79, Q&A: 0
    [Abstract] The retina is sensitive for light damages, because of direct light exposure, especially intense blue and UV light, which increase level of ROS and other toxic phototransduction products in photoreceptor cells. In our previous work (Damulewicz et al., 2017a and 2017b), we used 8-oxo-deoxyguanosine (8-OHdG) as a marker for oxidative stress ...
    Obtaining Acute Brain Slices
    Authors:  Thomas Papouin and Philip G. Haydon, date: 01/20/2018, view: 262, Q&A: 0
    [Abstract] Obtaining acute brain slices for electrophysiology or amperometric recordings has become a routine procedure in most labs in the field of neuroscience. Yet, protocols describing the step by step process are scarce, in particular for routine acute preparations such as from the mouse hippocampus. Here we provide a detailed protocol for the ...
    D-serine Measurements in Brain Slices or Other Tissue Explants
    Authors:  Thomas Papouin and Philip G. Haydon, date: 01/20/2018, view: 283, Q&A: 0
    [Abstract] D-serine is an atypical amino acid present in the mammalian body (most amino acids in the mammalian body are L-isomers) that is mostly known in neuroscience for its role as a co-agonist controlling the N-methyl D-aspartate receptor (NMDAR). D-serine levels are decreased in patients with schizophrenia and this is thought to mediate, at least in ...
    Mechanical Allodynia Assessment in a Murine Neuropathic Pain Model
    [Abstract] Experimental animal models are unique tools (i) to study pain transmission and pathophysiology of neuropathic pain, (ii) to identify novel molecular targets and (iii) to test the potential analgesic effect of specific molecules. The chronic constriction injury (CCI) model of neuropathic pain is the first model of post-traumatic painful peripheral ...
    Common Carotid Arteries Occlusion Surgery in Adult Rats as a Model of Chronic Cerebral Hypoperfusion
    Authors:  Dandan Cao, Yunfei Bai and Liang Li, date: 01/20/2018, view: 204, Q&A: 0
    [Abstract] Chronic cerebral hypoperfusion (CCH) is an important risk factor of vascular dementia (VaD) and Alzheimer’s disease (AD). Hypoxia/ischemia in the whole brain induced by CCH causes serious damage to brain structure and function, which can lead to cognitive impairment. Two-vessel occlusion (2-VO), also known as permanent, bilateral common carotid ...
    Fluorescent Measurement of Synaptic Activity Using FM Dyes in Dissociated Hippocampal Cultured Neurons
    Authors:  Roman M. Lazarenko, Claire E. DelBove and Qi Zhang, date: 01/20/2018, view: 218, Q&A: 0
    [Abstract] Release and recycling of synaptic vesicles are essential for neurotransmission and synaptic plasticity. To gain mechanistic understanding of these processes, direct measurements of vesicle release and retrieval is indispensable. Styryl dyes like FM1-43 and FM4-64 have been widely used for this purpose and their loading and unloading are reliable ...
    The RiboPuromycylation Method (RPM): an Immunofluorescence Technique to Map Translation Sites at the Sub-cellular Level
    Authors:  Amandine Bastide, Jonathan W. Yewdell and Alexandre David, date: 01/05/2018, view: 333, Q&A: 0
    [Abstract] While isotopic labeling of amino acids remains the reference method in the field for quantifying translation rate, it does not provide any information on spatial localization of translation sites. The rationale behind developing the ribopuromycylation method (RPM) was primarily to map translation sites at the sub-cellular level while avoiding ...
    A Rodent Model for Chronic Brain Hypoperfusion Related Diseases: Permanent Bilateral Occlusion of the Common Carotid Arteries (2VO) in Rats
    Authors:  Mei-Ling Yan and Jing Ai, date: 01/05/2018, view: 236, Q&A: 0
    [Abstract] Permanent occlusion of bilateral common carotid arteries (2VO) in rat is considered as a suitable animal model to mimic chronic brain hypoperfusion status, which is proved to be a risk factor to precede the Alzheimer’s disease or/and vascular dementia. In this protocol, we describe how to successfully ligate the bilateral common carotid arteries ...
    Mutant Huntingtin Secretion in Neuro2A Cells and Rat Primary Cortical Neurons
    Authors:  Katarina Trajkovic, Hyunkyung Jeong and Dimitri Krainc, date: 01/05/2018, view: 227, Q&A: 0
    [Abstract] Quantitative analysis of proteins secreted from the cells poses a challenge due to their low abundance and the interfering presence of a large amount of bovine serum albumin (BSA) in the cell culture media. We established assays for detection of mutant huntingtin (mHtt) secreted from Neuro2A cell line stably expressing mHtt and rat primary ...