flp-dependent mruby2 virus  (Jackson Laboratory)

Journal: IBRO Neuroscience Reports

Article Title: Co-transmitting neurons in the lateral septal nucleus exhibit features of neurotransmitter switching

doi: 10.1016/j.ibneur.2022.05.003

Figure Lengend Snippet: Genomic markers and viral labeling reveal ChAT/vGAT co-expressing population of neurons within the LSN. (A) Genomic or viral-based labeling of co-expressing neurons. (B) Location of the LSN. Green box indicates the location of the image shown in (C). Right, hemisphere of brain shown in (C). (C) ChAT/vGAT-positive neurons in the LSN labeled by tri-cistronic reporter. (D) Viral labeling of co-expressing LSN neurons. (E) Overlapping DIO-mVenus and fDIO-mRuby2 signals illustrating virally-targeted co-expressing neurons. (F) Quantified neurons within the LSN. Co-expressing neurons comprise 5% and 28% of vGAT-positive and ChAT-positive neurons, respectively. vGAT- and ChAT-positive neurons in each slice are normalized to the number of DAPI-positive nuclei counted in each slice. n = 28 images of the LSN from 1 animal.

Article Snippet: Assuming that subpopulations of vGAT LSN neurons would have connections within the LSN, we labeled local glutamatergic neurons by injecting a combination of Cre-dependent ChR2 (AAV-DIO-ChR2-EYFP) and Flp-dependent mRuby2 (AAV-fDIO-mRuby2) viruses into the LSN of vGAT-Cre; vGlut2-Flp (JAX Stock No. 030212) mice ( A-B).

Techniques: Labeling, Expressing

Journal: IBRO Neuroscience Reports

Article Title: Co-transmitting neurons in the lateral septal nucleus exhibit features of neurotransmitter switching

doi: 10.1016/j.ibneur.2022.05.003

Figure Lengend Snippet: LSN co-expressing neurons co-release ACh and GABA onto vGlut2-positive LSN neurons. (A) Photostimulation of ChR2-expressing vGAT-positive neurons while recording from mRuby2-expressing vGlut2-positive neurons in the LSN. (B) Confocal image of LSN with labeled channelrhodopsin-expressing vGAT-Cre-positive neurons (green) and vGlut2-Flp-positive neurons (red). (C) Recording of IPSCs in the presence of glutamatergic and GABAergic blockers. (D) Magnification of grey box in (C), IPSCs are resistant to bath application of mecamylamine, but sensitive to bath application of atropine.

Article Snippet: Assuming that subpopulations of vGAT LSN neurons would have connections within the LSN, we labeled local glutamatergic neurons by injecting a combination of Cre-dependent ChR2 (AAV-DIO-ChR2-EYFP) and Flp-dependent mRuby2 (AAV-fDIO-mRuby2) viruses into the LSN of vGAT-Cre; vGlut2-Flp (JAX Stock No. 030212) mice ( A-B).

Techniques: Expressing, Labeling