Assessing Neuron–Astrocyte Spatial Interactions Using the Neuron–Astrocyte Proximity Assay
| Autor: | Baljit S. Khakh, Mark H. G. Verheijen, J. Christopher Octeau, Aina Badia‐Soteras |
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| Přispěvatelé: | Amsterdam Neuroscience - Cellular & Molecular Mechanisms, Molecular and Cellular Neurobiology |
| Rok vydání: | 2020 |
| Předmět: |
Post hoc
Medium spiny neuron law.invention Synapse 03 medical and health sciences astrocyte Slice preparation synapse Confocal microscopy law optical medicine SDG 7 - Affordable and Clean Energy 030304 developmental biology 0303 health sciences Chemistry 030302 biochemistry & molecular biology imaging General Medicine neuron medicine.anatomical_structure Förster resonance energy transfer Neuron Neuroscience Astrocyte |
| Zdroj: | Badia-Soteras, A, Octeau, J C, Verheijen, M H G & Khakh, B S 2020, ' Assessing Neuron-Astrocyte Spatial Interactions Using the Neuron-Astrocyte Proximity Assay ', Current protocols in neuroscience, vol. 91, no. 1, e91, pp. e91 . https://doi.org/10.1002/cpns.91 Current protocols in neuroscience, 91(1):e91 |
| ISSN: | 1934-8576 1934-8584 |
| Popis: | Astrocytes are morphologically complex cells with numerous close contacts with neurons at the level of their somata, branches, and branchlets. The smallest astrocyte processes make discrete contacts with synapses at scales that cannot be observed by standard light microscopy. At such contact points, astrocytes are thought to perform both homeostatic and neuromodulatory roles-functions that are proposed to be determined by their close spatial apposition. To study such spatial interactions, we previously developed a Forster resonance energy transfer (FRET)-based approach, which enables observation and tracking of the static and dynamic proximity of astrocyte processes with synapses. The approach is compatible with standard imaging techniques such as confocal microscopy and permits assessment of the most proximate contacts between astrocytes and neurons in live tissues. In this protocol article we describe the approach to analyze the contacts between striatal astrocyte processes and corticostriatal neuronal projection terminals onto medium spiny neurons. We report the required protocols in detail, including adeno-associated virus microinjections, acute brain slice preparation, imaging, and post hoc FRET quantification. The article provides a detailed description that can be used to characterize and study astrocyte process proximity to synapses in living tissue. © 2020 by John Wiley & Sons, Inc. Basic Protocol 1: Forster resonance energy transfer imaging in cultured cells Basic Protocol 2: Forster resonance energy transfer imaging with the neuron-astrocyte proximity assay in acute brain slices. |
| Databáze: | OpenAIRE |
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