Localization of the glycine transporter GLYT1 in glutamatergic synaptic vesicles |
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| Institution: | 1. Centro de Biología Molecular “Severo Ochoa” and Departamento de Biología Molecular, Universidad Autónoma de Madrid-Consejo Superior de Investigaciones Científicas, Madrid, Spain;2. Centro de Investigación Biomédica en Red de Enfermedades Raras, Instituto de Salud Carlos III, Madrid, Spain;1. Center for Neuroscience, Department of Neurobiology, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15261, United States;2. Laboratory of Cellular and Molecular Neuroscience, National Institute of Mental Health, National Institute of Health, Bethesda, MD 20892, United States;1. Divisions of Gastroenterology and Hepatology, Keck School of Medicine of USC, Los Angeles, CA;2. Divisions of Gastroenterology and Hepatology, Washington University School of Medicine, St. Louis, Mo;3. Department of Medicine, St. Louis Health Care System, St. Louis, Mo;1. Institute of Pharmacology and Toxicology, University of Ulm Medical Center, Ulm, Germany;2. Rudolf-Virchow-Center, DFG-Research Center for Experimental Biomedicine, University of Würzburg, Würzburg, Germany;3. Institute of Organic Chemistry, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary;4. School of Engineering and Science, Jacobs University Bremen, Bremen, Germany;1. Department of Paediatrics, University of Calgary Faculty of Medicine and Alberta Children’s Hospital Research Institute, Calgary, Alberta, Canada;2. Department of Pathology (Neuropathology), University of Calgary Faculty of Medicine and Alberta Children’s Hospital Research Institute, Calgary, Alberta, Canada;3. Department of Pathology (Paediatric), University of Calgary Faculty of Medicine and Alberta Children’s Hospital Research Institute, Calgary, Alberta, Canada;4. Department of Clinical Neurosciences, University of Calgary Faculty of Medicine and Alberta Children’s Hospital Research Institute, Calgary, Alberta, Canada |
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| Abstract: | We have previously shown the presence of the glycine transporter GLYT1 in glutamatergic terminals of the rat brain. In this study we present immunohistochemical and biochemical evidence indicating that GLYT1 is expressed not only at the plasma membrane of glutamatergic neurons, but also at synaptic vesicles. Confocal microscopy, immunoblots analysis of a highly purified synaptic vesicle fraction and immunoisolation of synaptic vesicles with anti-synaptophysin antibodies strongly suggested the presence of GLYT1 in synaptic vesicles. Moreover, direct observation with the electron microscope of purified vesicles immunoreacted with anti-GLYT1 and colloidal gold demonstrated that about 40% of the small vesicles of the purified vesicle fraction contained GLYT1. Double labeling for GLYT1 and synaptophysin of this vesicular fraction revealed that more of ninety percent of them were synaptic vesicles. Moreover, a significant part of the GLYT1 containing vesicles (86%) also contained the vesicular glutamate transporter vGLUT1, suggesting a functional role of GLYT1 in a subpopulation of glutamatergic vesicles. |
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| Keywords: | Glycine Transporter GLYT1 Synaptic vesicles Glutamate |
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