A Knock-in Ntsr1-Flp Driver Enables Intersectional and Systemic Targeting of Heterogeneous Midbrain Dopamine Circuits
Year: 2026
Type: preprint
Abstract: Summary Precise genetic access to molecularly defined neuronal subpopulations is essential for dissecting circuit heterogeneity. We developed and validated a knock-in neurotensin receptor 1 ( Ntsr1 ) Flp mouse line for intersectional targeting of Ntsr1-expressing neurons. Following delivery of Flp-dependent adeno-associated viral (AAV) reporters, robust recombination was observed throughout the the midbrain and several other brain regions. A subset of Ntsr1 + neurons in the substantia nigra and ventral tegmental area lacked dopaminergic markers, indicating that midbrain Ntsr1 populations comprise both dopaminergic and non-dopaminergic neurons. Systemic delivery of a Cre and Flp-dependent reporter in complementary dual-recombinase configurations revealed configuration-dependent differences in dopaminergic targeting. Cis-gene controls ( Dat Cre ; Dat Flp ) defined the maximal dopaminergic targeting ceiling and helped distinguish true non-dopaminergic targeting from recombinase-dependent off-target labeling. Finally, a dual-recombinase-dependent taCaspase-3 construct enabled selective ablation of midbrain dopamine neurons in vivo, establishing Ntsr1 Flp as a versatile driver for scalable Boolean targeting.
Language: English
Subfield: Genetics
Domain: Life Sciences
Open Access status: green
Funders: California Institute of Technology, Michigan State University, National Institutes of Health, College of Engineering, Michigan State University, National Institute of Mental Health, National Institute of General Medical Sciences, National Institute of Diabetes and Digestive and Kidney Diseases
Awards: Broadening access with an Armamentarium Vector Core Powered by Inclusive Research Experiences, Identification of a dopamine circuit mediating day eating and diet-induced obesity in mice, Deciphering dopaminergic circuits required for food anticipatory activity in mice, Role of Lateral Hypothalamic Neurotensin Neurons in Energy Balance
OpenAlex ID: W7135021929