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

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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The IgSF Cell Adhesion Protein CLMP and Congenital Short Bowel Syndrome (CSBS)

International Journal of Molecular Sciences, Journal Year: 2023, Volume and Issue: 24(6), P. 5719 - 5719

Published: March 16, 2023

The immunoglobulin-like cell adhesion molecule CLMP is a member of the CAR family proteins and implicated in human congenital short-bowel syndrome (CSBS). CSBS rare but very severe disease for which no cure currently available. In this review, we compare data from patients mouse knockout model. These indicate that characterized by defect intestinal elongation during embryonic development impaired peristalsis. latter driven uncoordinated calcium signaling via gap junctions, linked to reduction connexin43 45 levels circumferential smooth muscle layer intestine. Furthermore, discuss how mutations gene affect other organs tissues, including ureter. Here, absence produces bilateral hydronephrosis—also caused reduced level associated junctions.

Language: Английский

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Lack of the Ig cell adhesion molecule BT-IgSF (IgSF11) induced behavioral changes in the open maze, water maze and resident intruder test

PLoS ONE, Journal Year: 2023, Volume and Issue: 18(1), P. e0280133 - e0280133

Published: Jan. 6, 2023

The brain- and testis-specific Ig superfamily protein (BT-IgSF, also termed IgSF11) is a homotypic cell adhesion protein. In the nervous system, BT-IgSF regulates stability of AMPA receptors in membrane cultured hippocampal neurons, modulates connectivity chandelier cells controls gap junction-mediated astrocyte-astrocyte communication. Here, we performed behavioral tests BT-IgSF-deficient mice. mice were similar to control littermates with respect their reflexes, motor coordination gating, associative learning. However, displayed an increased tendency stay central illuminated areas open field O-Maze paradigms suggesting reduced anxiety or scotophobia (fear darkness). Although initially found platform water maze behavior was compromised when moved, indicating flexibility. This deficit overcome by longer training improve spatial memory. Furthermore, male aggression towards intruder. Our results show that specific behaviors are modified lack demonstrate contribution network functions.

Language: Английский

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Membrane properties and coupling of macroglia in the optic nerve

Current Research in Neurobiology, Journal Year: 2024, Volume and Issue: 7, P. 100137 - 100137

Published: Jan. 1, 2024

We established a longitudinal acute slice preparation of transgenic mouse optic nerve to characterize membrane properties and coupling glial cells by patch-clamp dye-filling, complemented immunohistochemistry. Unlike in cortex or hippocampus, the majority EGFP + hGFAP-EGFP mouse, tool identify astrocytes, were characterized time voltage dependent K

Language: Английский

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