Temporal development of cyclic nucleotide-gated and Ca2+ -activated Cl- currents in isolated mouse olfactory sensory neurons

J Neurophysiol. 2007 Jul;98(1):153-60. doi: 10.1152/jn.00270.2007. Epub 2007 Apr 25.

Abstract

A Ca(2+)-activated Cl(-) current constitutes a large part of the transduction current in olfactory sensory neurons. The binding of odorants to olfactory receptors in the cilia produces an increase in cAMP concentration; Ca(2+) enters into the cilia through CNG channels and activates a Cl(-) current. In intact mouse olfactory sensory neurons little is known about the kinetics of the Ca(2+)-activated Cl(-) current. Here, we directly activated CNG channels by flash photolysis of caged cAMP or 8-Br-cAMP and measured the current response with the whole cell voltage-clamp technique in mouse neurons. We measured multiphasic currents in the rising phase of the response at -50 mV. The current rising phase became monophasic in the absence of extracellular Ca(2+), at +50 mV, or when most of the intracellular Cl(-) was replaced by gluconate to shift the equilibrium potential for Cl(-) to -50 mV. These results show that the second phase of the current in mouse intact neurons is attributed to a Cl(-) current activated by Ca(2+), similarly to previous results on isolated frog cilia. The percentage of the total saturating current carried by Cl(-) was estimated in two ways: 1) by measuring the maximum secondary current and 2) by blocking the Cl(-) channel with niflumic acid. We estimated that in the presence of 1 mM extracellular Ca(2+) and in symmetrical Cl(-) concentrations the Cl(-) component can constitute up to 90% of the total current response. These data show how to unravel the CNG and Ca(2+)-activated Cl(-) component of the current rising phase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 8-Bromo Cyclic Adenosine Monophosphate / pharmacology
  • Animals
  • Calcium / metabolism*
  • Chloride Channels / antagonists & inhibitors
  • Chloride Channels / physiology*
  • Cyclic AMP / analogs & derivatives
  • Cyclic AMP / pharmacology
  • Electric Stimulation / methods
  • Gluconates / pharmacology
  • Ion Channel Gating / drug effects
  • Ion Channel Gating / physiology*
  • Membrane Potentials / drug effects
  • Membrane Potentials / physiology
  • Membrane Potentials / radiation effects
  • Mice
  • Mice, Inbred BALB C
  • Neurons, Afferent / drug effects
  • Neurons, Afferent / physiology*
  • Niflumic Acid / pharmacology
  • Nucleotides, Cyclic / metabolism*
  • Nucleotides, Cyclic / pharmacology
  • Olfactory Mucosa / cytology*
  • Patch-Clamp Techniques / methods

Substances

  • (2-nitrophenyl)ethyl-cyclic adenosine monophosphate
  • Chloride Channels
  • Gluconates
  • Nucleotides, Cyclic
  • 8-Bromo Cyclic Adenosine Monophosphate
  • Niflumic Acid
  • Cyclic AMP
  • gluconic acid
  • Calcium