![]() Furthermore, astrocyte signaling can be investigated under physiological conditions, at high spatiotemporal resolution using in vivo two-photon (2P) microscopy and sensory-driven neuronal stimulation in awake animals ( 20). Now, automated, event-based analysis tools such as Automatic Quantification and Analysis (AQuA) ( 23) enable unbiased characterization of these spatially unfixed, size-varying Ca 2+ MDs, overcoming the limitations of traditional, region of interest (ROI)–based methods. The diversity of astrocytic Ca 2+ signals and their dynamics could explain how astrocytes can elicit such varied responses in the brain. Cells use the diverse spatiotemporal dynamics of i and the associated signaling network for versatile control of cellular processes ( 22). The spatiotemporal characteristics of these Ca 2+ MD signals are compatible with neuronal and vascular dynamics ( 9, 17– 21). Genetically encoded calcium indicators (GECIs) targeting astrocyte plasma membranes revealed fast and localized Ca 2+ microdomain (MD) activity in fine astrocytic processes. Technical advances offered solutions to these controversies and unearthed previously unidentified challenges. These functional subdomains are stable over days, suggesting subcellular specialization. The overlay of these signals resulted in behavior-dependent maps with characteristic Ca 2+ activity hotspots, maybe representing memory engrams. Frequency and size of signals were extensively increased by locomotion but only subtly with sensory stimulation. We found a wide range of Ca 2+ MD signals, some of which were ultrafast (≤300 ms). In combination with two-photon microscopy and unbiased, event-based analysis, we investigated cortical astrocytes embedded in the vibrissal thalamocortical circuit. Then, we use this axoastrocytic AAV transfer to express genetically encoded Ca 2+ indicators at high-contrast circuit specifically. Here, we first show that adeno-associated virus (AAV) particles transfer anterogradely from axons to astrocytes. However, functional organization of Ca 2+ MDs in space and time in relationship to behavior and neuronal activity is poorly understood. ![]() Astrocytes exhibit localized Ca 2+ microdomain (MD) activity thought to be actively involved in information processing in the brain. ![]()
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