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Sci. Signal., 9 November 2010
[DOI: 10.1126/scisignal.3147pe41]

Control of Breathing by "Nerve Glue" (Movies)

Klaus Ballanyi,* Bogdan Panaitescu, Araya Ruangkittisakul

*Corresponding author. E-mail: klaus.ballanyi{at}ualberta.ca

Description

These video animations relate to Fig. 2A in the main text and show multiphoton Ca2+ imaging of inspiratory active neurons and smaller, unidentified glial cells in the inspiratory rhythm-generating center, the pre-Bötzinger complex (preBötC), from a 700 μm-thick, anatomically "calibrated", brainstem slice from a 1-day-old rat in which the preBötC is exposed to the caudal surface (1). The preBötC is pressure-injected with 0.5 mM membrane-permeant Ca2+-sensitive fluorescent dye, Fluo4-AM for monitoring dynamic changes in the free cytosolic Ca2+ concentration ("Ca2+ activity") (2).

Movie 1. Ca2+ oscillations in neurons of the preBötC. A population of preBötC neurons shows somatic Ca2+ oscillations that are synchronous with inspiratory rhythmic bursting recorded with a suction electrode from the contralateral preBötC. Large-amplitude synchronous Ca2+ rises (due to "fictive sighs") are followed by more frequent smaller-amplitude signals (due to "fictive eupnea" bursts). Small cells, presumably astrocytes, do not show rhythmic Ca2+ increases that are in phase with inspiratory activity, although they can generate spontaneous (non–respiratory-related) Ca2+ rises that are much larger in amplitude than inspiratory oscillations. This spontaneous Ca2+ activity is apparent in one putative astrocyte in the top middle section of the image. The video captures a time period of 100 s.

[Play Movie 1]

Movie 2. Morphology of neurons and glial cells in the rhythmic preBötC slice. A "Z-stack animation" of the preBötC region from Movie 1 shows the morphology of neurons and glial cells as the focus image plane is moved from 35 to 85 μm below the surface of the slice. The optical recording is performed during bath application of 0.5 mM glutamate so as to induce a persistent increase in Ca2+ for better visualization of cell morphology. Most neurons have a fusiform soma shape, and their processes and their proximal dendrites are visible. Astrocytes are smaller (~10 μm soma diameter) and have a rounder shape. Scale is the same as in Movie 1.

[Play Movie 2]

References

1. A. Ruangkittisakul, S.W. Schwarzacher, L. Secchia, Y. Ma, N. Bobocea, B.Y. Poon, G. D. Funk, K. Ballanyi, Generation of eupnea and sighs by a spatiochemically organized inspiratory network. J. Neurosci. 28, 2447–2458 (2008).

2. A. Ruangkittisakul, Y. Okada, Y. Oku, N. Koshiya, K. Ballanyi, Fluorescence imaging of active respiratory networks. Resp. Physiol. Neurobiol. 168, 26–38 (2009).

Technical Details

Format: AVI

Size: 4.7 MB (Movie 1); 6.3 MB (Movie 2)

Requirements: Windows Media Player or QuickTime player


Citation: K. Ballanyi, B. Panaitescu, A. Ruangkittisakul, Control of Breathing by "Nerve Glue". Sci. Signal. 3, pe41 (2010).

© 2010 American Association for the Advancement of Science


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