Feasibility of electrical recordings from unconnected vertebrate CNS neurons cultured in a three-dimensional extracellular matrix |
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| Authors: | P W Coates R D Nathan |
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| Affiliation: | 1. Department of Cell Biology and Anatomy, Texas Tech University Health Sciences Center, School of Medicine, Lubbock, TX 79430 U.S.A.;2. Department of Physiology, Texas Tech University Health Sciences Center, School of Medicine, Lubbock, TX 79430 U.S.A.;1. Program in Neurogenetics, Department of Neurology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA;2. F.M. Kirby Neurobiology Center, Boston Children’s Hospital and Harvard Medical School, Boston, MA 02115, USA;3. Interdepartmental Program in Neuroscience, University of California, Los Angeles, Los Angeles, CA 90095, USA;4. Department of Neurosciences, University of California, San Diego, La Jolla, CA 92093, USA;5. Department of Biological Chemistry, Weizmann Institute of Science, 76100 Rehovot, Israel;6. Department of Human Genetics, University of California, Los Angeles, Los Angeles, CA 90095, USA;7. Department of Biostatistics, University of California, Los Angeles, Los Angeles, CA 90095, USA;8. Department of Psychiatry, Semel Institute for Neuroscience and Human Behavior, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA 90095, USA;1. BIOMATEN, Middle East Technical University (METU) Center of Excellence in Biomaterials and Tissue Engineering, Ankara, Turkey;2. METU, Department of Biomedical Engineering, Ankara, Turkey;3. METU, Department of Biotechnology, Ankara, Turkey;4. METU, Department of Biological Sciences, Ankara, Turkey;1. Northeastern University, Department of Chemical Engineering, Boston, MA, 02115, United States;2. Department of Biomedical Engineering, University of Miami, Coral Gables, FL, 33146, United States;3. Department of Biology, Boston, 02115, MA, United States;1. Department of Biomaterials Engineering, Faculty of Health Sciences, UCAM-Universidad Católica San Antonio de Murcia, Guadalupe, 30107 Murcia, Spain;2. Center of Molecular Medicine and Diagnostics (COMManD), Department of Biochemistry, Saveetha Dental College and Hospitals, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai 600 077, India;3. Department of Marine Biopharmacology, College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China;4. Green and Innovative Technologies for Food, Environment and Bioengineering Research Group, Faculty of Pharmacy and Nutrition, UCAM-Universidad Católica San Antonio de Murcia, 30107 Murcia, Spain;5. Universidad San Francisco de Quito USFQ, Politecnico, Quito 170901, Ecuador;6. Department of Implant Dentistry, Faculty of Medicine and Dentistry, UCAM-Universidad Católica San An-tonio de Murcia, 30107 Murcia, Spain |
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| Abstract: | Single, i.e. unconnected spinal or cerebral neurons from chick embryos rapidly regenerate morphologically identifiable axons and dendrites when cultured in a three-dimensional (3-D) extracellular matrix (ECM) consisting of a hydrated native collagen lattice. We now show that it is possible to study the intrinsic electrophysiological properties of such neurons as early as 2 days in culture. Cells were plated at 10(5) cells/dish using Medium 199 containing 10% fetal calf serum but no other supplements or antibiotics-antimycotics. A patch-clamp/whole-cell voltage clamp system was used to record single-channel currents from cell-attached patches, transmembrane potentials during the injection of rectangular currents, and whole-cell currents during voltage clamp. After 2-4 days in culture, isolated cerebral and spinal neurons exhibited single-channel currents. Within 7 days, rectangular currents injected through the recording electrode evoked action potentials. These results demonstrate that unconnected CNS neurons quickly display at least some properties of excitability when cultured in a 3-D ECM. This culture system should facilitate investigation of intrinsic electrical properties of single CNS neurons, and how extrinsic factors including neurotransmitters, hormones, pharmacological agents and contacts with other cells influence electrical activity. |
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