Effects of calretinin on Ca2+ signals in cerebellar granule cells: implications of cooperative Ca2+ binding

Calretinin is thought to be the main endogenous calcium buffer in cerebellar granule cells (GrCs). However, little is known about the impact of cooperative Ca(2+) binding to calretinin on highly localized and more global (regional) Ca(2+) signals in these cells. Using numerical simulations, we show that an essential property of calretinin is a delayed equilibration with Ca(2+). Therefore, the amount of Ca(2+), which calretinin can accumulate with respect to equilibrium levels, depends on stimulus conditions. Based on our simulations of buffered Ca(2+) diffusion near a single Ca(2+) channel or a large cluster of Ca(2+) channels and previous experimental findings that 150 μM 1,2-bis(o-aminophenoxy) ethane-N, N, N', N'-tetraacetic acid (BAPTA) and endogenous calretinin have similar effects on GrC excitability, we estimated the concentration of mobile calretinin in GrCs in the range of 0.7-1.2 mM. Our results suggest that this estimate can provide a starting point for further analysis. We find that calretinin prominently reduces the action potential associated increase in cytosolic free Ca(2+) concentration ([Ca(2+)]( i )) even at a distance of 30 nm from a single Ca(2+) channel. In spite of a buildup of residual Ca(2+), it maintains almost constant maximal [Ca(2+)]( i ) levels during repetitive channel openings with a frequency less than 80 Hz. This occurs because of accelerated Ca(2+) binding as calretinin binds more Ca(2+). Unlike the buffering of high Ca(2+) levels within Ca(2+) nano/microdomains sensed by large conductance Ca(2+)-activated K(+) channels, the buffering of regional Ca(2+) signals by calretinin can never be mimicked by certain concentration of BAPTA under all different experimental conditions.