Fluorescent Tagging of Rhythmically Active Respiratory Neurons within the Pre-Botzinger Complex of Rat Medullary Slice Preparations

Elucidation of the neuronal mechanisms underlying respiratory rhythmogenesis is a major focal point in respiratory physiology. An area of the ventrolateral medulla, the pre-Bötzinger complex (preBotC), is a critical site. Attention is now focused on understanding the cellular and network properties...

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Bibliographic Details
Published in:The Journal of neuroscience 2005-03, Vol.25 (10), p.2591-2596
Main Authors: Pagliardini, Silvia, Adachi, Tadafumi, Ren, Jun, Funk, Gregory D, Greer, John J
Format: Article
Language:English
Subjects:
Animals
Brief Communications
Fluorescent Dyes - analysis
In Vitro Techniques
Isotope Labeling - methods
Medulla Oblongata - chemistry
Medulla Oblongata - physiology
Molecular Probe Techniques
Molecular Probes - analysis
Neurons - chemistry
Neurons - physiology
Periodicity
Rats
Rats, Sprague-Dawley
Receptors, Neurokinin-1 - analysis
Receptors, Neurokinin-1 - physiology
Respiration
Respiratory Mechanics - physiology
Substance P - analysis
Substance P - physiology
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Summary:Elucidation of the neuronal mechanisms underlying respiratory rhythmogenesis is a major focal point in respiratory physiology. An area of the ventrolateral medulla, the pre-Bötzinger complex (preBotC), is a critical site. Attention is now focused on understanding the cellular and network properties within the preBotC that underlie this critical function. The inability to clearly identify key "rhythm-generating" neurons within the heterogeneous population of preBotC neurons has been a significant limitation. Here we report an advancement allowing precise targeting of neurons expressing neurokinin-1 receptors (NK1Rs), which are hypothesized to be essential for respiratory rhythmogenesis. The internalization of tetramethylrhodamine conjugated substance P in rhythmically active medullary slice preparations provided clear visualization of NK1R-expressing neurons for subsequent whole-cell patch-clamp recordings. Among labeled neurons, 82% were inspiratory modulated, and 25% had pacemaker properties. We propose that this approach can be used to greatly expedite progress toward understanding the neuronal processes underlying the control of breathing.
ISSN:0270-6474
1529-2401
DOI:10.1523/JNEUROSCI.4930-04.2005