Immediate responses to ambient light in vivo reveal distinct subpopulations of suprachiasmatic VIP neurons.

The circadian rhythm pacemaker, the suprachiasmatic nucleus (SCN), mediates light entrainment via vasoactive intestinal peptide (VIP) neurons (SCNVIP). Yet, how these neurons uniquely respond and connect to intrinsically photosensitive retinal ganglion cells (ipRGCs) expressing melanopsin (Opn4) has not been determined functionally in freely behaving animals. To address this, we first used monosynaptic tracing from SCNVIP neurons in mice and identified two SCNVIP subpopulations. Second, we recorded calcium changes in response to ambient light, at both bulk and single-cell levels, and found two unique activity patterns in response to high- and low-intensity blue light. The activity patterns of both subpopulations could be manipulated by application of an Opn4 antagonist. These results suggest that the two SCNVIP subpopulations connect to two types of Opn4-expressing ipRGCs, likely M1 and M2, but only one is responsive to red light. These findings have important implications for our basic understanding of non-image-forming circadian light processing.

A neural mechanism for discriminating social threat from social safety.

The ability to distinguish a threatening from non-threatening conspecific based on past experience is critical for adaptive social behaviors. Although recent progress has been made in identifying the neural circuits that contribute to different types of positive and negative social interactions, the neural mechanisms that enable the discrimination of individuals based on past aversive experiences remain unknown. Here, we developed a modified social fear conditioning paradigm that induced in both sexes robust behavioral discrimination of a conspecific associated with a footshock (CS+) from a non-reinforced interaction partner (CS-). Strikingly, chemogenetic or optogenetic silencing of hippocampal CA2 pyramidal neurons, which have been previously implicated in social novelty recognition memory, resulted in generalized avoidance fear behavior towards the equally familiar CS-and CS+. One-photon calcium imaging revealed that the accuracy with which CA2 representations discriminate the CS+ from the CS-animal was enhanced following social fear conditioning and strongly correlated with behavioral discrimination. Moreover the CA2 representations incorporated a generalized or abstract representation of social valence irrespective of conspecific identity and location. Thus, our results demonstrate, for the first time, that the same hippocampal CA2 subregion mediates social memories based on conspecific familiarity and social threat, through the incorporation of a representation of social valence into an initial representation of social identity.

Light-guided sectioning for precise in situ localization and tissue interface analysis for brain-implanted optical fibers and GRIN lenses.

Optical implants to control and monitor neuronal activity in vivo have become foundational tools of neuroscience. Standard two-dimensional histology of the implant location, however, often suffers from distortion and loss during tissue processing. To address that, we developed a three-dimensional post hoc histology method called "light-guided sectioning" (LiGS), which preserves the tissue with its optical implant in place and allows staining and clearing of a volume up to 500 µm in depth. We demonstrate the use of LiGS to determine the precise location of an optical fiber relative to a deep brain target and to investigate the implant-tissue interface. We show accurate cell registration of ex vivo histology with single-cell, two-photon calcium imaging, obtained through gradient refractive index (GRIN) lenses, and identify subpopulations based on immunohistochemistry. LiGS provides spatial information in experimental paradigms that use optical fibers and GRIN lenses and could help increase reproducibility through identification of fiber-to-target localization and molecular profiling.

Anatomy and activity patterns in a multifunctional motor neuron and its surrounding circuits.

Dorsal Excitor motor neuron DE-3 in the medicinal leech plays three very different dynamical roles in three different behaviors. Without rewiring its anatomical connectivity, how can a motor neuron dynamically switch roles to play appropriate roles in various behaviors? We previously used voltage-sensitive dye imaging to record from DE-3 and most other neurons in the leech segmental ganglion during (fictive) swimming, crawling, and local-bend escape (Tomina and Wagenaar, 2017). Here, we repeated that experiment, then re-imaged the same ganglion using serial blockface electron microscopy and traced DE-3's processes. Further, we traced back the processes of DE-3's presynaptic partners to their respective somata. This allowed us to analyze the relationship between circuit anatomy and the activity patterns it sustains. We found that input synapses important for all the behaviors were widely distributed over DE-3's branches, yet that functional clusters were different during (fictive) swimming vs. crawling.