Second messenger molecules have a limited spread in olfactory cilia

Abstract

Odorants are detected by olfactory receptors on the sensory cilia of olfactory receptor cells (ORCs). These cylindrical cilia have a diameters of 100–200 nm, within which the components required for signal transduction by the adenylyl cyclase–cAMP system are located. The kinetics of odorant responses are determined by the lifetimes of active proteins as well as the production, diffusion, and extrusion/degradation of second messenger molecules (cAMP and Ca2+). However, there is limited information about the molecular kinetics of ORC responses, mostly because of the technical limitations involved in studying such narrow spaces and fine structures. In this study, using a combination of electrophysiology, photolysis of caged substances, and spot UV laser stimulation, we show that second messenger molecules work only in the vicinity of their site of generation in the olfactory cilia. Such limited spreading clearly explains a unique feature of ORCs, namely, the integer multiple of unitary events that they display in low Ca2+ conditions. Although the small ORC uses cAMP and Ca2+ for various functions in different regions of the cell, these substances seem to operate only in the compartment that has been activated by the appropriate stimulus. We also show that these substances remain in the same vicinity for a long time. This enables the ORC to amplify the odorant signal and extend the lifetime of Ca2+-dependent adaptation. Cytoplasmic buffers and extrusion/degradation systems seem to play a crucial role in limiting molecular spreading. In addition, binding sites on the cytoplasmic surface of the plasma membrane may limit molecular diffusion in such a narrow space because of the high surface/volume ratio. Such efficient energy conversion may also be broadly used in other biological systems that have not yet been subjected to systematic experiments.