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J Appl Physiol 89: 807-822, 2000;
8750-7587/00 $5.00
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Vol. 89, Issue 2, 807-822, August 2000

INNOVATIVE TECHNIQUES
Continuous intracellular recording from mammalian neurons exposed to hyperbaric helium, oxygen, or air

Jay B. Dean and Daniel K. Mulkey

Department of Physiology and Biophysics, College of Science and Mathematics, Wright State University School of Medicine, Dayton, Ohio 45435

We developed a hyperbaric chamber for intracellular recording in rat brain stem slices during continuous compression and decompression of the tissue bath with the inert gas helium. Air, rather than helium, was also used as the compression medium in some cases to increase tissue nitrogen levels. An important feature is the chamber door, which opens or closes rapidly at 1 atmosphere absolute (ATA) for increased accessibility of the microelectrode. The door also closes and seals smoothly without disrupting the intracellular recording. Hyperbaric oxygen was administered during helium compression using a separate pressure cylinder filled with perfusate equilibrated with 2.3-3.3 ATA oxygen. Measurements of tissue/bath PO2 and pH confirmed that the effects of compression using helium or air could be differentiated from those due to increased PO2. One hundred and thirteen neurons were studied during 375 compression cycles ranging from 1 to 20 ATA (mode 3.0 ATA). We conclude that it is technically feasible to record intracellularly from the same mammalian neuron while changing ambient pressure over a physiologically important range. These techniques will be useful for studying how various hyperbaric environments affect neurophysiological mechanisms.

diving medicine; hyperbaric medicine; inert gas narcosis; oxygen toxicity; reactive oxygen species


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