No abstract

Oxytocin (OXT) is a neuropeptide originating in the paraventricular nucleus (PVN) of the hypothalamus, with a role in influencing various social behaviors. However, pinpointing its actions only during the time animals are performing specific behaviors has been difficult to study. Here we developed an optogenetic gene expression system designed to selectively label neuronal populations activated by OXT in the presence of blue-light, named OXTR-iTango2. The OXTR-iTango2 was capable of inducing gene expression of a reporter gene in both human embryonic kidney (HEK) cells and neurons in a quantitative manner. In vivo expression of OXTR-iTango2 selectively labeled OXT-sensitive neurons in a blue-light dependent manner. Furthermore, we were able to detect a subset of dopamine (DA) neurons in the ventral tegmental area (VTA) that receive OXT activation during social interaction. Thus, we provide a genetically-encoded, scalable optogenetic toolset to target neural circuits activated by OXT in behaving animals with a high temporal resolution.

Verifying causal effects of neural circuits is essential for proving a direct circuit-behavior relationship. However, techniques for tagging only active neurons with high spatiotemporal precision remain at the beginning stages. Here we develop the soma-targeted Cal-Light (ST-Cal-Light) which selectively converts somatic calcium rise triggered by action potentials into gene expression. Such modification simultaneously increases the signal-to-noise ratio of reporter gene expression and reduces the light requirement for successful labeling. Because of the enhanced efficacy, the ST-Cal-Light enables the tagging of functionally engaged neurons in various forms of behaviors, including context-dependent fear conditioning, lever-pressing choice behavior, and social interaction behaviors. We also target kainic acid-sensitive neuronal populations in the hippocampus which subsequently suppress seizure symptoms, suggesting ST-Cal-Light’s applicability in controlling disease-related neurons. Furthermore, the generation of a conditional ST-Cal-Light knock-in mouse provides an opportunity to tag active neurons in a region- or cell-type specific manner via crossing with other Cre-driver lines. Thus, the versatile ST-Cal-Light system links somatic action potentials to behaviors with high temporal precision, and ultimately allows functional circuit dissection at a single cell resolution.