Abstract

.Significance: Insights into the cellular activity of each member of the neurovascular unit (NVU) is critical for understanding their contributions to neurovascular coupling (NVC)—one of the key control mechanisms in cerebral blood flow regulation. Advances in imaging and genetic tools have enhanced our ability to observe, manipulate and understand the cellular activity of NVU components, namely neurons, astrocytes, microglia, endothelial cells, vascular smooth muscle cells, and pericytes. However, there are still many unresolved questions. Since astrocytes are considered electrically unexcitable, signaling is the main parameter used to monitor their activity. It is therefore imperative to study astrocytic dynamics simultaneously with vascular activity using tools appropriate for the question of interest.Aim: To highlight currently available genetic and imaging tools for studying the NVU—and thus NVC—with a focus on astrocyte dynamics and vascular activity, and discuss the utility, technical advantages, and limitations of these tools for elucidating NVC mechanisms.Approach: We draw attention to some outstanding questions regarding the mechanistic basis of NVC and emphasize the role of astrocytic elevations in functional hyperemia. We further discuss commonly used genetic, and optical imaging tools, as well as some newly developed imaging modalities for studying NVC at the cellular level, highlighting their advantages and limitations.Results: We provide an overview of the current state of NVC research, focusing on the role of astrocytic elevations in functional hyperemia; summarize recent advances in genetically engineered indicators, fluorescence microscopy techniques for studying NVC; and discuss the unmet challenges for future imaging development.Conclusions: Advances in imaging techniques together with improvements in genetic tools have significantly contributed to our understanding of NVC. Many pieces of the puzzle have been revealed, but many more remain to be discovered. Ultimately, optimizing NVC research will require a concerted effort to improve imaging techniques, available genetic tools, and analytical software.

Highlights

  • Neurovascular coupling (NVC) is an essential control mechanism in the regulation of cerebral blood flow (CBF)

  • We provide an overview of the current state of NVC research, focusing on the role of astrocytic Ca2þ elevations in functional hyperemia; summarize recent advances in genetically engineered Ca2þ indicators, fluorescence microscopy techniques for studying NVC; and discuss the unmet challenges for future imaging development

  • Advances in imaging techniques together with improvements in genetic tools have significantly contributed to our understanding of NVC

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Summary

Results

We provide an overview of the current state of NVC research, focusing on the role of astrocytic Ca2þ elevations in functional hyperemia; summarize recent advances in genetically engineered Ca2þ indicators, fluorescence microscopy techniques for studying NVC; and discuss the unmet challenges for future imaging development

Conclusions
Introduction
Neurovascular Coupling
Direct Neuronal Activation of Blood Vessels
Astrocytes Relay Neuronal-Activation Status to Blood Vessels
Optical Imaging Tools for In Vivo Studies
Future Opportunities
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