Isolation,Culture and Long-Term Maintenance of Primary Mesencephalic Dopaminergic Neurons From Embryonic Rodent Brains |
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Authors: | Maria Weinert Tharakeswari Selvakumar Travis S Tierney Kambiz N Alavian |
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Affiliation: | 1.Division of Brain Sciences, Department of Medicine, Imperial College London;2.Department of Neurosurgery, Brigham and Women''s Hospital, Harvard Medical School;3.Department of Internal Medicine, Endocrinology, Yale University School of Medicine |
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Abstract: | Degeneration of mesencephalic dopaminergic (mesDA) neurons is the pathological hallmark of Parkinson’s diseae. Study of the biological processes involved in physiological functions and vulnerability and death of these neurons is imparative to understanding the underlying causes and unraveling the cure for this common neurodegenerative disorder. Primary cultures of mesDA neurons provide a tool for investigation of the molecular, biochemical and electrophysiological properties, in order to understand the development, long-term survival and degeneration of these neurons during the course of disease. Here we present a detailed method for the isolation, culturing and maintenance of midbrain dopaminergic neurons from E12.5 mouse (or E14.5 rat) embryos. Optimized cell culture conditions in this protocol result in presence of axonal and dendritic projections, synaptic connections and other neuronal morphological properties, which make the cultures suitable for study of the physiological, cell biological and molecular characteristics of this neuronal population. |
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Keywords: | Neuroscience Issue 96 Ventral midbrain Parkinson''s disease Dopaminergic Primary neuronal culture Neuronal development Neurodegeneration |
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