Kynurenate treatment of autaptic hippocampal microcultures affect localized voltage-dependent calcium diffusion in the dendrites

It is not clear how different spatial compartments in the neuron are affected during epileptiform activity. In the present study we have examined the spatial and temporal profiles of depolarization induced changes in the intracellular Ca2+ concentration in the dendrites of cultured autaptic hippocampal pyramidal neurons rendered epileptic experimentally by treatment with kynurenate (2 mM) and Mg2+ (11.3 mM) in culture (treated neurons). This was examined with simultaneous somatic patch-pipette recording and Ca2+ imaging experiments using the Ca2+ indicator Oregon Green 488 BAPTA-1. Neurons stimulated by depolarization under whole-cell voltage clamp conditions revealed Ca2+ entry at localized sites in the dendrites. Ca2+ transients were observed even in the presence of NMDA and AMPA receptor antagonists suggesting that the opening of voltage gated calcium channels primarily triggered the local Ca2+ changes. Peak Ca2+ transients in the dendrites of treated neurons were larger compared to the signals recorded from the control neurons. Dendritic Ca2+ transients in treated neurons showed a distance dependent scaling. Estimation of dendritic local Ca2+ diffusion coefficients indicated higher values in the treated neurons and a higher availability of free Ca2+. Simulation studies of Ca2+ dynamics in these localized dendritic compartments indicate that local Ca2+ buffering and removal mechanisms may be affected in treated neurons. Our studies indicate that small dendritic compartments are rendered more vulnerable to changes in intracellular Ca2+ following induction of epileptiform activity. This can have important cellular consequences including local membrane excitability through mechanisms that remain to be elucidated.