Pathway-specific inputs to the superior colliculus support flexible responses to visual threat.

Behavioral flexibility requires directing feedforward sensory information to appropriate targets. In the superior colliculus, divergent outputs orchestrate different responses to visual threats, but the circuit organization enabling the flexible routing of sensory information remains unknown. To determine this structure, we focused on inhibitory projection (Gad2) neurons. Trans-synaptic tracing and neuronal recordings revealed that Gad2 neurons projecting to the lateral geniculate nucleus (LGN) and the parabigeminal nucleus (PBG) form two separate populations, each receiving a different set of non-retinal inputs. Inhibiting the LGN- or PBG-projecting Gad2 neurons resulted in opposing effects on behavior; increasing freezing or escape probability to visual looming, respectively. Optogenetic activation of selected inputs to the LGN- and PBG-projecting Gad2 cells predictably regulated responses to visual threat. These data suggest that projection-specific sampling of brain-wide inputs provides a circuit design principle that enables visual inputs to be selectively routed to produce context-specific behavior.

Optogenetic fUSI for brain-wide mapping of neural activity mediating collicular-dependent behaviors.

Neuronal cell types are arranged in brain-wide circuits that guide behavior. In mice, the superior colliculus innervates a set of targets that direct orienting and defensive actions. We combined functional ultrasound imaging (fUSI) with optogenetics to reveal the network of brain regions functionally activated by four collicular cell types. Stimulating each neuronal group triggered different behaviors and activated distinct sets of brain nuclei. This included regions not previously thought to mediate defensive behaviors, for example, the posterior paralaminar nuclei of the thalamus (PPnT), which we show to play a role in suppressing habituation. Neuronal recordings with Neuropixels probes show that (1) patterns of spiking activity and fUSI signals correlate well in space and (2) neurons in downstream nuclei preferentially respond to innately threatening visual stimuli. This work provides insight into the functional organization of the networks governing innate behaviors and demonstrates an experimental approach to explore the whole-brain neuronal activity downstream of targeted cell types.

A Platform for Brain-wide Volumetric Functional Ultrasound Imaging and Analysis of Circuit Dynamics in Awake Mice.

Imaging large-scale circuit dynamics is crucial to understanding brain function, but most techniques have a limited depth of field. Here, we describe volumetric functional ultrasound imaging (vfUSI), a platform for brain-wide vfUSI of hemodynamic activity in awake head-fixed mice. We combined a high-frequency 1,024-channel 2D-array transducer with advanced multiplexing and high-performance computing for real-time 3D power Doppler imaging at a high spatiotemporal resolution (220 × 280 × 175 µm3, up to 6 Hz). We developed a standardized software pipeline for registration, segmentation, and temporal analysis in 268 individual brain regions based on the Allen Mouse Common Coordinate Framework. We demonstrated the high sensitivity of vfUSI under multiple experimental conditions, and we successfully imaged stimulus-evoked activity when only a few trials were averaged. We also mapped neural circuits in vivo across the whole brain during optogenetic activation of specific cell types. Moreover, we identified the sequential activation of sensory-motor networks during a grasping water-droplet task.

Septal GABAergic inputs to CA1 govern contextual memory retrieval.

The CA1 output region of the hippocampus plays an essential role in the retrieval of episodic memories. γ-Aminobutyric acid-releasing (GABAergic) long-range projections from the medial septum (MS) densely innervate the hippocampus, but whether septal inputs regulate memory expression remains elusive. We found that the MS to CA1 connection is recruited during recall of a contextual fear memory. Chemogenetic silencing of CA1-projecting MS neurons or septal GABAergic terminals within CA1 blocked memory retrieval. Photostimulation of septal GABAergic terminals in CA1 selectively inhibited interneurons. Abrogating septal GABAergic cells during retrieval disinhibited parvalbumin-rich (PV+) cells in CA1. Direct activation of CA1 PV+ cells impaired memory and prevented the induction of extracellular signal-regulated kinase/mitogen-activated kinase signaling in postsynaptic pyramidal neurons. Opposing disinhibition of hippocampal PV+ cells reversibly restored memory. Our data indicate that suppression of feed-forward inhibition onto CA1 by septal GABAergic neurons is an important mechanism in gating contextual fear behavior.