Mapping multi-regional functional connectivity of astrocyte-neuronal networks during behaviors.

Significance: Diverse behaviors rely on coordinated activity and multi-regional functional connectivity within astrocyte-neuronal networks. However, current techniques for simultaneously measuring astrocytic and neuronal activities across multiple brain regions during behaviors remain limited.

Aim: We propose a multi-fiber solution that can simultaneously record activities of astrocyte-neuronal networks across multiple regions during behaviors.

Approach: We employed cell-specific dual-color genetically encoded calcium indicators (GECIs) and multi-fiber photometry to simultaneously measure astrocytic and neuronal Ca transients across multiple brain regions in freely behaving animals.

Results: Our findings demonstrate that both movements and sensory stimuli induce synchronized and highly correlated Ca transients in astrocytes and neurons of freely behaving mice. In addition, we recorded astrocytic and neuronal Ca transients from multiple brain regions during mouse behaviors. Our observations reveal heightened synchronization of astrocytic and neuronal Ca transients across different brain regions during movements or sensory stimuli, indicating enhanced functional connectivity within brain-wide astrocyte-neuronal networks.

Conclusions: Multi-fiber photometry, combined with cell-specific dual-color GECIs, represents a powerful approach for investigating astrocytic and neuronal activities across different brain regions during behaviors. This technique serves as a versatile tool for analyzing the multi-regional functional connectivity map of astrocyte-neuronal networks associated with specific behaviors.