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Gap junctions in the neonatal mouse pre‐Bötzinger complex: Insights using fluorescence recovery after photobleaching
Author(s) -
Kelty Jonathan,
Shields Edward
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.1088.6
Subject(s) - carbenoxolone , photobleaching , gap junction , fluorescence recovery after photobleaching , biophysics , fluorescence , chemistry , neuron , neurite , anatomy , neuroscience , biology , intracellular , biochemistry , in vitro , membrane , optics , physics
Whether interneuronal coupling via gap junctions is essential to the generation/patterning of rhythmic activity by an inspiration‐related network in the medulla's pre‐Bötzinger complex remains unresolved. We used a fluorescence recovery after photobleaching approach in medullary slice preparations from neonatal mice to determine whether diffusion of carboxyfluorescein from neuron to neuron is blocked by the gap junction uncoupler carbenoxolone (CBX). Tissues incubated in standard artificial cerebrospinal fluid (aCSF) regained a greater percentage of their initial fluorescence than did those incubated in aCSF + CBX. Regardless of whether slices were incubated in aCSF or in aCSF + CBX, almost all bleached somata regained some fluorescence within 20 min following photobleaching. Presumably this recovery was due, at least in part, to movement of carboxyfluorescein into somata from adjacent portions of neurites. Only a small fraction of bleached somata directly abutted other neuronal somata exhibiting fluorescence suggesting that either a) carboxyfluorescein moved into recovering somata from adjacent glia and/or b) that CBX somehow interferes with the redistribution of carboxyfluorescein within cells.