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Journal of Neuroscience, Vol 11, 3573-3580, Copyright © 1991 by Society for Neuroscience
Analysis of single cyclic nucleotide-gated channels in olfactory receptor cells
F Zufall, S Firestein and GM Shepherd
Physiological Institute, Technical University Munich, Germany.
In the accompanying article (Firestein et al., 1991b), we have demonstrated
that odor- and cyclic nucleotide-sensitive channels exist at a low density
in the dendritic membranes of isolated salamander olfactory receptor
neurons. Here, we analyze the cyclic nucleotide sensitivity of these
channels using the inside-out patch recording technique. Both cAMP and
cGMP, at micromolar concentrations, are capable of inducing channel
openings. The biophysical parameters of channel activity are nearly the
same in response to either ligand. The unitary conductance is about 45 pS,
the reversal potential of single- channel currents is +5 mV, and the I/V
relation is linear over the range -80 to +80 mV. The channel activity shows
no obvious voltage dependence in divalent cation-free symmetrical
solutions. The channel shows no desensitization, even to agonist exposures
lasting 15 sec. Mean open time is about 1.5 msec; the closed time
distribution is best fit by two exponentials with a fast time constant in
the submillisecond range (ca. 0.15 msec) and a slower time constant in the
millisecond range (ca. 1.5 msec). The only clear difference in the activity
of the two ligands is in their affinity constants. The K1/2 for cAMP is 20
microM; that for cGMP is 4 microM. In both cases, the Hill coefficient is
greater than 2, suggesting that channel opening requires the cooperative
action of three ligand molecules.
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