Cyclic nucleotide-gated (CNG) channels are tetrameric nonspecific cation channels. They mediate the receptor potentials in photoreceptors and cells of the olfactory epithelium and they are activated by the binding of cyclic nucleotides, such as cGMP and cAMP. Prior studies in homotetrameric CNGA1 channels, activated with covalently bound cGMP, presented evidence that partially liganded channels cause partial channel opening (Ruiz and Karpen, 1997,1999). Here, the conductance and the gating of homotetrameric CNGA1 channels are studied as function of the concentration of freely diffusible cGMP and with different permeating ions. At saturating cGMP the current levels distributed around a single mean in a Gaussian fashion and the open times were long. At low cGMP, however, the current levels were heterogenous: They were smaller than those at saturating cGMP, equal, or larger. The open times were short. Ions generating the larger single-channel currents (Na⁺>K⁺>Rb⁺) concomitantly increased the heterogeneity of current levels and decreased the open probability and the open times. The results suggest that activation of CNGA1 channels by cGMP and ions staying longer in the pore is associated with less extensive and less frequent conformational fluctuations of the channel pore.