Visualization of Gap Junction–Mediated Astrocyte Coupling in Acute Mouse Brain Slices DOI Creative Commons

Nine Kompier,

Gabrielle Siemonsmeier,

Niklas Meyer

et al.

BIO-PROTOCOL, Journal Year: 2025, Volume and Issue: 15(1365)

Published: Jan. 1, 2025

Gap junctions are transmembrane protein channels that enable the exchange of small molecules such as ions, second messengers, and metabolites between adjacent cells. found in various mammalian organs, including skin, endothelium, liver, pancreas, muscle, central nervous system (CNS). In CNS, they mediate coupling neural cells glial cells, resulting panglial networks vital for brain homeostasis. Tracers sufficiently molecular mass can diffuse across gap used to visualize extent cell-to-cell situ by delivering them a single cell through sharp electrodes or patch-clamp micropipettes. Here, we describe protocol pre-labeling identification astrocytes acute mouse forebrain slices using Sulforhodamine 101 (SR101). Fluorescent then be targeted whole-cell patch-clamp, which allows further confirmation astroglial identity assessing their electrophysiological properties, well passive dialysis with tracer biocytin. Slices subjected chemical fixation immunostaining detect dye-coupled networks. This provides method live tissue SR101 labeling. Alternatively, transgenic reporter mice also identify astrocytes. While illustrate use this study brain, general principles applicable other species, tissues, types. Key features • Pre-labeling adult dye 101. Dialysis biocytin into individual electrophysiology. Staining streptavidin GFAP, imaging, analysis astrocytic Can types might adapted tissues species.

Language: Английский

Visualization of Gap Junction–Mediated Astrocyte Coupling in Acute Mouse Brain Slices DOI Creative Commons

Nine Kompier,

Gabrielle Siemonsmeier,

Niklas Meyer

et al.

BIO-PROTOCOL, Journal Year: 2025, Volume and Issue: 15(1365)

Published: Jan. 1, 2025

Gap junctions are transmembrane protein channels that enable the exchange of small molecules such as ions, second messengers, and metabolites between adjacent cells. found in various mammalian organs, including skin, endothelium, liver, pancreas, muscle, central nervous system (CNS). In CNS, they mediate coupling neural cells glial cells, resulting panglial networks vital for brain homeostasis. Tracers sufficiently molecular mass can diffuse across gap used to visualize extent cell-to-cell situ by delivering them a single cell through sharp electrodes or patch-clamp micropipettes. Here, we describe protocol pre-labeling identification astrocytes acute mouse forebrain slices using Sulforhodamine 101 (SR101). Fluorescent then be targeted whole-cell patch-clamp, which allows further confirmation astroglial identity assessing their electrophysiological properties, well passive dialysis with tracer biocytin. Slices subjected chemical fixation immunostaining detect dye-coupled networks. This provides method live tissue SR101 labeling. Alternatively, transgenic reporter mice also identify astrocytes. While illustrate use this study brain, general principles applicable other species, tissues, types. Key features • Pre-labeling adult dye 101. Dialysis biocytin into individual electrophysiology. Staining streptavidin GFAP, imaging, analysis astrocytic Can types might adapted tissues species.

Language: Английский

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