The Probability of Triggering Calcium Puffs Is Linearly Related to the Number of Inositol Trisphosphate Receptors in a Cluster
2012
Dickinson, George D. | Swaminathan, Divya | Parker, Ian
Puffs are local Ca²⁺ signals that arise by Ca²⁺ liberation from the endoplasmic reticulum through concerted opening of tightly clustered inositol trisphosphate receptor/channels (IP₃R). They serve both local signaling functions and trigger global Ca²⁺ waves. The numbers of functional IP₃R within clusters differ appreciably between different puff sites, and we investigated how the probability of puff occurrence varies with cluster size. We imaged puffs in SH-SY5Y cells using total internal fluorescence microscopy, and estimated cluster sizes from the magnitude of the largest puff observed at each site relative to the signal from a single channel. We find that the initial triggering rate of puffs following photorelease of IP₃, and the average frequency of subsequent repetitive puffs, vary about linearly with cluster size. These data accord well with stochastic simulations in which opening of any individual IP₃R channel within a cluster triggers a puff via Ca²⁺-induced Ca²⁺ release. An important consequence is that the signaling power of a puff site (average amount of Ca²⁺ released per puff × puff frequency) varies about the square of cluster size, implying that large clusters contribute disproportionately to cellular signaling and, because of their higher puff frequency, preferentially act as pacemakers to initiate Ca²⁺ waves.
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