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Local Ca2+ detection and modulation of synaptic release by astrocytes

Nature Neuroscience volume 14, pages 1276–1284 (2011)Cite this article

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Abstract

Astrocytes communicate with synapses by means of intracellular calcium ([Ca2+]i) elevations, but local calcium dynamics in astrocytic processes have never been thoroughly investigated. By taking advantage of high-resolution two-photon microscopy, we identify the characteristics of local astrocyte calcium activity in the adult mouse hippocampus. Astrocytic processes showed intense activity, triggered by physiological transmission at neighboring synapses. They encoded synchronous synaptic events generated by sparse action potentials into robust regional (∼12 μm) [Ca2+]i elevations. Unexpectedly, they also sensed spontaneous synaptic events, producing highly confined (∼4 μm), fast (millisecond-scale) miniature Ca2+ responses. This Ca2+ activity in astrocytic processes is generated through GTP- and inositol-1,4,5-trisphosphate–dependent signaling and is relevant for basal synaptic function. Thus, buffering astrocyte [Ca2+]i or blocking a receptor mediating local astrocyte Ca2+ signals decreased synaptic transmission reliability in minimal stimulation experiments. These data provide direct evidence that astrocytes are integrated in local synaptic functioning in adult brain.

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Figure 1: Imaging Ca2+ activity in micrometric astrocytic process domains in the adult hippocampus.
Figure 2: Two types of Ca2+ events, focal and expanded, in astrocytic processes.
Figure 3: Ca2+ events in astrocytic processes are triggered by spontaneous and action potential–dependent synaptic activity.
Figure 4: Local synaptic release evoked by sucrose triggers focal Ca2+ events in astrocytic processes.
Figure 5: Ca2+ events in astrocytic processes are mediated by GTP- and InsP3-dependent signaling.
Figure 6: Blockade of Ca2+ activity in astrocytic processes decreases basal transmission at local synapses: role of P2Y1R.

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Acknowledgements

We thank P. Bezzi for experiments in cultured astrocytes and critical comments, K.D. McCarthy (University of North Carolina, Chapel Hill) and P.G. Haydon (Tufts University) for providing Itpr2−/− mice, H. Kettenmann (Max Delbrück Center) for providing GFAP-EGFP mice, V. Crunelli, R. Schneggenburger and F. Helmchen for suggestions, and T. Oertner for critical reading of a previous manuscript version. This work was supported by grants from Swiss National Science Foundation (3100A0–100850 and 3100A0–120398) to A.V.; J.C. received a postdoctoral fellowship from MBF Foundation, Triesen, LI/Synapsis Foundation, Zurich; P.T., from Swiss National Science Foundation National Center of Competence in Research 'Synapsy'. M.S. received a University of Lausanne Faculty of Biology and Medicine PhD fellowship.

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  1. Maria Amalia Di Castro and Julien Chuquet: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Cell Biology and Morphology, University of Lausanne, Lausanne, Switzerland

    Maria Amalia Di Castro, Julien Chuquet, Nicolas Liaudet, Khaleel Bhaukaurally, Mirko Santello, David Bouvier, Pascale Tiret & Andrea Volterra

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Contributions

M.A.D.C. performed imaging, electrophysiology (including minimal stimulations) and data analysis; J.C. coordinated imaging, software development and performed imaging and data analysis; and the three next authors contributed equally: N.L. developed software for imaging data analysis and participated in imaging and imaging data analysis; K.B. performed imaging and electrophysiology and contributed to software development in the first part of the project; and M.S. performed imaging, electrophysiology and electrophysiology analysis. D.B. performed immunohistochemistry, P.T. performed imaging and data analysis in the last part of the project, and A.V. coordinated the project and wrote the manuscript.

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Correspondence to Andrea Volterra.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–13, Supplementary Data 1 and 2 (PDF 722 kb)

Supplementary Video 1

Ca2+ activity in astrocytic processes in basal condition. Ca2+ imaging sequence (150 s, 3 Hz, accelerated 5 times) in the processes of an astrocyte in the adult dentate molecular layer. Ca2+ activity comprises “focal” Ca2+ events, randomly recurrent, fast local transients; and “expanded” Ca2+ events, less frequent, large scale, longer-lasting transients. (AVI 2535 kb)

Supplementary Video 2

From astrocytes to micrometric sub-regions: the segmentation principle. The movie summarizes the different steps of segmentation of astrocytic processes into sub-regions of ~1 μm+ in which Ca2+ activity is studied. 3D morphology is built from the fluorescence signal of the Ca2+-insensitive dye Texas Red. (AVI 1720 kb)

Supplementary Video 3

Effect of TTX on Ca2+ activity in astrocytic processes. Dual sequence (150 s, 3 Hz) of Ca2+ activity in basal condition (left) and upon TTX treatment (right). Blocking action potentials affects primarily expanded Ca2+ events, whereas focal Ca2+ activity is largely maintained. (AVI 1979 kb)

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Di Castro, M., Chuquet, J., Liaudet, N. et al. Local Ca2+ detection and modulation of synaptic release by astrocytes. Nat Neurosci 14, 1276–1284 (2011). https://doi.org/10.1038/nn.2929

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