In Utero Electroporation Approaches to Study the Excitability of Neuronal Subpopulations and Single-cell Connectivity
| Autor: | Marta Nieto, Carlos G. Briz, Marta Navarrete, José A. Esteban |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Nervous system education.field_of_study General Immunology and Microbiology Electroporation General Chemical Engineering General Neuroscience Population Biology Corpus callosum General Biochemistry Genetics and Molecular Biology Cortex (botany) Coupling (electronics) 03 medical and health sciences Electrophysiology 030104 developmental biology medicine.anatomical_structure nervous system medicine Patch clamp education Neuroscience |
| Zdroj: | Journal of Visualized Experiments. |
| ISSN: | 1940-087X |
| DOI: | 10.3791/55139-v |
| Popis: | The nervous system is composed of an enormous range of distinct neuronal types. These neuronal subpopulations are characterized by, among other features, their distinct dendritic morphologies, their specific patterns of axonal connectivity, and their selective firing responses. The molecular and cellular mechanisms responsible for these aspects of differentiation during development are still poorly understood. Here, we describe combined protocols for labeling and characterizing the structural connectivity and excitability of cortical neurons. Modification of the in utero electroporation (IUE) protocol allows the labeling of a sparse population of neurons. This, in turn, enables the identification and tracking of the dendrites and axons of individual neurons, the precise characterization of the laminar location of axonal projections, and morphometric analysis. IUE can also be used to investigate changes in the excitability of wild-type (WT) or genetically modified neurons by combining it with whole-cell recording from acute slices of electroporated brains. These two techniques contribute to a better understanding of the coupling of structural and functional connectivity and of the molecular mechanisms controlling neuronal diversity during development. These developmental processes have important implications on axonal wiring, the functional diversity of neurons, and the biology of cognitive disorders. |
| Databáze: | OpenAIRE |
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