Estimating intracellular Ca2+ concentrations and buffering in a dendritic inhibitory hippocampal interneuron.
Calcium is known to regulate several phenomena like neuronal excitability and plasticity. Interestingly, the spatiotemporal profile of dendritic calcium depends on several processes, specific to each neuronal type. In this study, we investigated Ca(2+) buffering and action potential (AP)-evoked Ca(2+) signaling in the dendrites of anatomically identified oriens lacunosum-moleculare (O-LM) cells, a major type of dendrite-targeting interneurons in the hippocampal CA1 region, using a combination of whole-cell patch-clamp recording and fast Ca(2+) imaging in acute rat brain slices. Cells were loaded with fluorescent Ca(2+) indicators fura-2 or Oregon Green BAPTA-1 (OGB-1) via patch-clamping electrode, and the effect of fura-2 on AP-evoked dendritic Ca(2+) transients was determined by ratiometric Ca(2+) imaging. To estimate intracellular Ca(2+) concentrations ([Ca(2+)](i)) and endogenous Ca(2+)-binding ratio (kappa(s)) in the proximal dendrite, fluorescence signals were converted into [Ca(2+)](i) using the ratioing method and were analyzed on the basis of the "single compartment model." Resting [Ca(2+)](i) was 22+/-5 nM and the build-up of [Ca(2+)](i) during a single AP was up to 656+/-226 nM. Analysis of Ca(2+) transients revealed that O-LM cells have a relatively low endogenous Ca(2+)-binding ratio (kappa(s)): the kappa(s) was 20+/-8 estimated during fura-2 loading and 27 estimated under steady-state fura-2 concentrations, respectively. To further examine the spatial profile of dendritic Ca(2+) transients, we measured somatic AP-evoked Ca(2+) transients beyond proximal dendrites using OGB-1. Dendritic Ca(2+) transients evoked by single APs or AP trains are not limited to regions close to the soma. The amplitude and decay of [Ca(2+)](i) associated with backpropagating APs are relatively independent of the distance from the soma. In sum, O-LM cells exhibit low endogenous Ca(2+)-binding ratios and relatively distance-independent Ca(2+) dynamics in the dendrites. These special features of Ca(2+) signaling in O-LM cells may have important functional implications for both normal and pathological conditions.
Institute of Neuroscience and Brain Research Center, National Yang-Ming University, 155, Section 2, Li-Nong Street, Taipei 112, Taiwan, Republic of China.
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Lien
D000200:Action Potentials; D000814:Aniline Compounds; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D003713:Dendritic Cells / Q000378:metabolism*; D005452:Fluoresceins; D005456:Fluorescent Dyes; D016257:Fura-2; D006624:Hippocampus / Q000378:metabolism*; D066298:In Vitro Techniques; D007395:Interneurons / Q000378:metabolism*; D042541:Intracellular Space / Q000378:metabolism; D008297:Male; D018408:Patch-Clamp Techniques; D011485:Protein Binding; D051381:Rats; D017207:Rats, Sprague-Dawley
D000814:Aniline Compounds; D005452:Fluoresceins; D005456:Fluorescent Dyes; C503942:Oregon green 488 BAPTA-1; D002118:Calcium; D016257:Fura-2