Hindered cytoplasmic diffusion of inositol trisphosphate restricts its cellular range of action

The range of action of intracellular messengers is determined by their rates of diffusion and degradation. Previous measurements in oocyte cytoplasmic extracts indicated that the Ca²⁺-liberating second messenger inositol trisphosphate (IP₃) diffuses with a coefficient (~280 μm² s^-1) similar to that in water, corresponding to a range of action of ~25 μm. Consequently, IP₃ is generally considered a "global" cellular messenger. We reexamined this issue by measuring local IP₃-evoked Ca²⁺ puffs to monitor IP₃ diffusing from spot photorelease in neuroblastoma cells. Fitting these data by numerical simulations yielded a diffusion coefficient (≤10 μm² s^-1) about 30-fold slower than that previously reported. We propose that diffusion of IP₃ in mammalian cells is hindered by binding to immobile, functionally inactive receptors that were diluted in oocyte extracts. The predicted range of action of IP₃ (<5 μm) is thus smaller than the size of typical mammalian cells, indicating that IP₃ should better be considered as a local rather than a global cellular messenger.