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Diffusion and Extrusion Shape Standing Calcium Gradients During Ongoing Parallel Fiber Activity in Dendrites of Purkinje Neurons

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Abstract

Synaptically induced calcium transients in dendrites of Purkinje neurons (PNs) play a key role in the induction of plasticity in the cerebellar cortex (Ito, Physiol Rev 81:1143–1195, 2001). Long-term depression at parallel fiber–PN synapses can be induced by stimulation paradigms that are associated with long-lasting (>1 min) calcium signals. These signals remain strictly localized (Eilers et al., Learn Mem 3:159–168, 1997), an observation that was rather unexpected, given the high concentration of the mobile endogenous calcium-binding proteins parvalbumin and calbindin in PNs (Fierro and Llano, J Physiol (Lond) 496:617–625, 1996; Kosaka et al., Exp Brain Res 93:483–491, 1993). By combining two-photon calcium imaging experiments in acute slices with numerical computer simulations, we found that significant calcium diffusion out of active branches indeed takes places. It is outweighed, however, by rapid and powerful calcium extrusion along the dendritic shaft. The close interplay of diffusion and extrusion defines the spread of calcium between active and inactive dendritic branches, forming a steep gradient in calcium with drop ranges of ~13 μm (interquartile range, 10–18 μm).

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Acknowledgements

We thank Klaus Stiefel for help in preliminary simulations and Gudrun Bethge for technical assistance.

Conflict of Interest Statement

The authors declare that there is no conflict of interest.

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  1. Carl-Ludwig-Institute for Physiology, University Leipzig, Leipzig, Germany

    Hartmut Schmidt, Oliver Arendt & Jens Eilers

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  1. Hartmut Schmidt
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Correspondence to Jens Eilers.

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Fig. S1

Localization of the Ca2+ plateaus with respect to branchpoints. The distance of the 50% value of the Ca2+ plateau to the nearest distal and proximal branchpoints (circles, connected by a straight line) plotted for each individual experiment. The data are spatially normalized to the points of inflexion of the logistic fit to the data, indicated by the dashed line in the scheme on the top. The left gray segment denotes the active dendritic compartment, the right segment the inactive one (cf. inset to Fig. 2d). The median of the measured drop ranges is indicated by the shading and the bracket. Note that in distal dendrites of PNs, branching occurs approximately every 10 μm (Ito M. The cerebellum and neuronal control. New York: Raven Press; 1984; cf. figure 1b). (PDF 466 kb)

Fig. S2

Influence of a side branch on the steady-state Ca2+ gradient. a Simulation of the drop in Ca2+ between the 30-μm long active dendritic segment coupled to the 50 μm long inactive segment (horizontal segments in the schematic illustration in the inset; 2 μm diameter) and to an additional 30-μm long inactive side branch (vertical segment) located at the center of the active segment. The diameter of the side branch was varied from 0.2 to 1.0 and 2.0 μm diameter (dotted, dashed, and broken line, respectively). The solid line represents the simulation in the absence of the side branch, corresponding to the solid lines in the one-dimensional simulations shown in Figs. 2d, 3a, and 4a. b Same as in a but with setting the side branch to “active” (i.e., to harboring a plateau [Ca2+] i of 295 nM). c–f Same as in a and b but for a side branch connected to the intersection between the primary active and inactive dendritic segments (c and d) or to the inactive segment (15 μm distant from the horizontal intersection; e and f) (PDF 380 kb)

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Schmidt, H., Arendt, O. & Eilers, J. Diffusion and Extrusion Shape Standing Calcium Gradients During Ongoing Parallel Fiber Activity in Dendrites of Purkinje Neurons. Cerebellum 11, 694–705 (2012). https://doi.org/10.1007/s12311-010-0246-x

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