title: Mutational analysis of dendritic Ca2+ kinetics in rodent Purkinje cells: role of parvalbumin and calbindin D28k.
abstract: The mechanisms governing the kinetics of climbing fibre-mediated Ca2+ transients in spiny dendrites of cerebellar Purkinje cells (PCs) were quantified with high-resolution confocal Ca2+ imaging. Ca2+ dynamics in parvalbumin (PV-/-) and parvalbumin/calbindin D28k null-mutant (PV/CB-/-) mice were compared with responses in wild-type (WT) animals. In the WT, Ca2+ transients in dendritic shafts were characterised by double exponential decay kinetics that were not due to buffered Ca2+ diffusion or saturation of the indicator dye. Ca2+ transients in PV-/- PCs reached the same peak amplitude as in the WT but the biphasic nature of the decay was less pronounced, an effect that could be attributed to PV's slow binding kinetics. In contrast, peak amplitudes in PV/CB-/- PCs were about two times higher than in the WT and the decay became nearly monophasic. Numerical simulations indicate that the residual deviation from a single exponential decay in PV/CB-/- is due to saturation of the Ca2+ indicator dye. Furthermore, the simulations imply that the effect of uncharacterised endogenous Ca2+ binding proteins is negligible, that buffered diffusion and dye saturation significantly affects spineous Ca2+ transients but not those in the dendritic shafts, and that neither CB nor PV undergoes saturation in spines or dendrites during climbing fibre-evoked Ca2+ transients. Calbindin's medium-affinity binding sites are fast enough to reduce the peak amplitude of the Ca2+ signal. However, similar to PV, delayed binding by CB leads to biphasic Ca2+ decay kinetics. Our results suggest that the distinct kinetics of PV and CB underlie the biphasic kinetics of synaptically evoked Ca2+ transients in dendritic shafts of PCs.
authors.affiliation: Department of Neurophysiology, Max-Planck-Institute for Brain Research, 60528 Frankfurt, Germany.
authors.forename: Hartmut
authors.initials: H
authors.lastname: Schmidt
authors.forename: Klaus M
authors.initials: KM
authors.lastname: Stiefel
authors.forename: Peter
authors.initials: P
authors.lastname: Racay
authors.forename: Beat
authors.initials: B
authors.lastname: Schwaller
authors.forename: Jens
authors.initials: J
authors.lastname: Eilers
mesh_terms: D000818:Animals; D064092:Calbindin 1; D064026:Calbindins; D002118:Calcium / Q000378:metabolism* / Q000502:physiology; D020013:Calcium Signaling; D002135:Calcium-Binding Proteins / Q000502:physiology; D004396:Coloring Agents; D004252:DNA Mutational Analysis; D003712:Dendrites / Q000378:metabolism*; D004058:Diffusion; D004594:Electrophysiology; D005453:Fluorescence; D066298:In Vitro Techniques; D007425:Intracellular Membranes / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D018345:Mice, Knockout; D018613:Microscopy, Confocal; D008863:Microspheres; D009412:Nerve Fibers / Q000502:physiology; D009994:Osmolar Concentration; D010320:Parvalbumins / Q000172:deficiency / Q000378:metabolism / Q000502:physiology*; D011689:Purkinje Cells / Q000378:metabolism*; D064030:S100 Calcium Binding Protein G / Q000378:metabolism / Q000502:physiology*
chemical_list: C576499:Calb1 protein, mouse; D064092:Calbindin 1; D064026:Calbindins; D002135:Calcium-Binding Proteins; D004396:Coloring Agents; D010320:Parvalbumins; D064030:S100 Calcium Binding Protein G; D002118:Calcium