yfp channel  (Revvity)

Journal: Neuroscience

Article Title: Pituitary Adenylate Cyclase Activating Polypeptide Inhibits A 10 Dopamine Neurons and Suppresses the Binge-like Consumption of Palatable Food.

doi: 10.1016/j.neuroscience.2021.09.016

Figure Lengend Snippet: Fig. 1. Coronal schematics illustrating the accuracy of our bilateral AAV ChR2 injections in the VMN (A) and the guide cannula implantations and AAV eYFP injections in the VTA (B).

Article Snippet: AAV constructs containing either cation channel rhodopsin-2 (ChR2) (pAAV-Ef1a-DIO hChR2(E123A)-E YFP.YFP.WPRE.jGH; 7.2 1012 genomic copies/mL; 300 nL total volume; a gift from Karl Deisseroth; Addgene plasmid #35507) or its enhanced yellow fluorescent protein (eYFP) blank control (pAAV-Ef1aDIO EYFP; 1.0 1013; 300 nL total volume; Addgene plasmid #27056), as well as the retrograde tracer Fluorogold (Conde et al., 2016) (Fluorochrome, LLC, Denver, Colo.,USA; 0.2 lL) were injected bilaterally over 2 min. We kept the injection needle in place for 10 min to enable adequate diffusion from the tip, after which we slowly retracted it from the brain.

Techniques:

Journal: Neuroscience

Article Title: Pituitary Adenylate Cyclase Activating Polypeptide Inhibits A 10 Dopamine Neurons and Suppresses the Binge-like Consumption of Palatable Food.

doi: 10.1016/j.neuroscience.2021.09.016

Figure Lengend Snippet: Fig. 5. Photostimulation of VMN PACAP neurons inhibits VTA neurons in PACAP-Cre mice. (A, B) eYFP ChR2 reporter signal in the VMN (4X & 40X). (C, D) eYFP ChR2 reporter signal in the VTA (4X & 40X). (E) DIC image (40X) of a recorded VTA neuron. (F–I) Optogenetic stimulation of VMN PACAP neurons produces an outward current associated with an increased slope conductance and reversal of polarity at -90 mV in the vast majority of VTA neurons (n = 15). Arrows indicate where I/Vs were conducted.

Article Snippet: AAV constructs containing either cation channel rhodopsin-2 (ChR2) (pAAV-Ef1a-DIO hChR2(E123A)-E YFP.YFP.WPRE.jGH; 7.2 1012 genomic copies/mL; 300 nL total volume; a gift from Karl Deisseroth; Addgene plasmid #35507) or its enhanced yellow fluorescent protein (eYFP) blank control (pAAV-Ef1aDIO EYFP; 1.0 1013; 300 nL total volume; Addgene plasmid #27056), as well as the retrograde tracer Fluorogold (Conde et al., 2016) (Fluorochrome, LLC, Denver, Colo.,USA; 0.2 lL) were injected bilaterally over 2 min. We kept the injection needle in place for 10 min to enable adequate diffusion from the tip, after which we slowly retracted it from the brain.

Techniques: