A novel dopamine transporter transgenic mouse line for identification and purification of midbrain dopaminergic neurons reveals midbrain heterogeneity

• DOI: 10.1111/ejn.13046
• Publication Date: 2015-08-19T08:32:50Z
• Authors: Mia Apuschkin; Sara Stilling; Troels Rahbek‐Cle...; Gunnar Sørensen; Guillaume Fortin; Freja Herborg Hansen; Jacob Eriksen; Louis‐Eric Trudeau; Kristoffer Egerod; Ulrik Gether; Mattias Rickhag
• Keywords: Male; 0301 basic medicine; Dopamine Plasma Membrane Transport Proteins; Behavior, Animal; Tyrosine 3-Monooxygenase; Dopaminergic Neurons; Green Fluorescent Proteins; Ventral Tegmental Area; Mice, Transgenic; Flow Cytometry; Mice, Inbred C57BL; Mice; 03 medical and health sciences; Animals; Female; Synaptosomes

Abstract Midbrain dopaminergic ( DA ergic) neurons are a heterogeneous cell group, composed of functionally distinct populations projecting to the basal ganglia, prefrontal cortex and limbic system. Despite their functional significance, midbrain population ergic is sparse, constituting only 20 000–30 000 in mice, development novel tools identify these cells warranted. Here, bacterial artificial chromosome mouse line [ Dat1 ‐enhanced green fluorescent protein eGFP )] from Gene Expression Nervous System Atlas GENSAT ) that expresses under control dopamine transporter DAT promoter was characterized. Confocal microscopy analysis brain sections showed strong e GFP signal reporter regions striatal terminals co‐localized with markers tyrosine hydroxylase TH ). Thorough quantification co‐localization ventral vast majority ‐expressing ergic. Importantly, expression profiles also revealed heterogeneity when comparing substantia nigra tegmental area. ‐e mice neither change synaptosomal uptake nor altered levels both striatum midbrain. No behavioural difference between Dat 1‐ wild‐type found, suggesting strain not aberrant. Finally, highly enriched can be obtained postnatal by fluorescence‐activated sorting sorted cultured vitro . The current investigation demonstrates this selective for neurons, constitutes promising tool delineating new aspects biology.