Journal: The Journal of Neuroscience
Article Title: Three Types of Neurochemical Projection from the Bed Nucleus of the Stria Terminalis to the Ventral Tegmental Area in Adult Mice
Figure Lengend Snippet: Postsynaptic receptor phenotype in the VTA. A , Triple immunofluorescence for GFP (green), GABA A Rα1 (red), and TH (blue) in the VTA of GAD67–GFP knock-in mice. GABA A Rα1 is expressed selectively in GFP-immunopositive GABAergic neurons (asterisk). B , C , Double-labeling preembedding immunoelectron microscopy for BDA (diffuse DAB precipitates) and GABA A Rα1 (metal particles) in the VTA of wild-type mice. At BST–VTA symmetrical synapses (enlarged in C , white arrowheads), GABA A Rα1 is localized on the synaptic and extrasynaptic membranes of dendrites (arrows). D , Postembedding immunoelectron microscopy for GABA A Rα1 (ϕ = 10 nm) and VIAAT (ϕ = 15 nm). VIAAT-labeled terminals form symmetrical synapses (white arrowheads), and GABA A Rα1 subunit is localized on the postsynaptic membrane (white arrow). E , Postembedding immunoelectron microscopy for GluA1 (ϕ = 10 nm) and VGluT2 (ϕ = 15 nm). VGluT2-labeled terminals form asymmetrical synapses (black arrowheads), and GluA1 is localized on the postsynaptic membrane (black arrows). Scale bars: A , 30 μm; B , 500 nm; C–E , 200 nm. Dn, Dendrite; NT, nerve terminals.
Article Snippet: Digoxigenin (DIG)- or fluorescein-labeled cRNA probes were prepared to detect multiple mRNAs simultaneously. cDNA fragments of GAD67 [1036–2015; National Center for Biotechnology Information (NCBI) Reference Sequence ], mouse VGluT1 (301–1680; NCBI Reference Sequence ), mouse VGluT2 (934–2060; NCBI Reference Sequence ), and mouse VGluT3 (22–945; NCBI Reference Sequence ) were subcloned into the Bluescript II plasmid vector.
Techniques: Immunofluorescence, Knock-In, Mouse Assay, Labeling, Immuno-Electron Microscopy