Activation of cyclic nucleotide-gated (CNG) channels represents the final step in the transduction pathways in both vision and olfaction. Over the past several years, CNG channels have been found in a variety of other cell types where they might fulfill various physiological functions. The olfactory and photoreceptor CNG channels rely on the binding of at least two molecules of cAMP or cGMP at intracellular sites on the channel protein to open a nonspecific cation conductance with a significant permeability to Ca ions. A series of elegant experiments with cloned channels and chimeric constructs has revealed significant information regarding the binding and gating reactions that lead to CNG channel activation. These recent studies have identified several regions as well as specific amino acid residues distributed on the retinal or the olfactory CNG channel subunits that play a key role in channel regulation. In this review, we will focus on these specific molecular sites of activation and modulation of CNG channels.