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Am J Physiol Cell Physiol 280: C1387-C1393, 2001;
0363-6143/01 $5.00
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Vol. 280, Issue 6, C1387-C1393, June 2001

Noninvasive measurement of chloride concentration in rat olfactory receptor cells with use of a fluorescent dye

Hiroshi Kaneko1, Tadashi Nakamura1,2, and Bernd Lindemann3

1 Department of Applied Physics and Chemistry, Division of Bio-Informatics, 2 Department of Information Network Science, The University of Electro-Communications, Chofu, Tokyo 182 - 8585, Japan; and 3 Department of Physiology, Saar University, D-66421 Homburg, Germany

Inwardly directed Ca2+-dependent chloride currents are thought to prolong and boost the odorant-induced transient receptor currents in olfactory cilia. Cl- inward current, of course, requires a sufficiently high intracellular Cl- concentration ([Cl-]i). In previous measurements using a fluorescent Cl- probe, N-(ethoxycarbonylmethyl)-6-methoxyquinolinium bromide (MQAE), [Cl-]i of newt olfactory cells was estimated to be only 40 mM. This low value led us to reexamine the [Cl-]i by an improved procedure. When isolated rat olfactory neurons were bathed in Tyrode's solution (150 mM Cl-) at room temperature, the [Cl-] was 81.5 ± 13.5 mM (mean ± SE) in the tip of the dendrite (olfactory knob) and 81.8 ± 10.2 mM (mean ± SE) in the soma. The corresponding Cl- equilibrium potentials were -15.4 and -15.3 mV, respectively. Therefore, at resting potentials in the range of -90 to -50 mV, Cl- currents are predicted to be inward and capable of contributing to the depolarization induced by odorants. Yet, if the cell was depolarized beyond -15 mV, somal Cl- currents would be outward and facilitate repolarization during excitation. The measured [Cl-] in soma and knob are of interest, because in the cilia the chloride content may be expected to equilibrate with that of the knob in the resting state. They provide a starting point for the decrease in ciliary [Cl-] predicted to occur during transduction.

Ca2+-gated Cl- channel; reversal potential; imaging; N-(ethoxycarbonylmethyl)-6-methoxyquinolinium bromide


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