raav-hsyn-egfp #50465-aavrg  (Addgene inc)

Journal: Nature Communications

Article Title: Brainstem noradrenergic modulation of the kisspeptin neuron GnRH pulse generator in mice

doi: 10.1038/s41467-025-60837-8

Figure Lengend Snippet: a Experimental design for excitatory and inhibitory DREADD studies. Retrograde mCherry-tagged excitatory (hM3Dq) or inhibitory (hM4Di) DREADD AAVs were stereotaxically injected into the ARN alongside placement of an optic fiber for GCaMP photometry in Kiss1 Cre/+ ,Dbh FLP/+ ,Ai162 +/+ mice . Changes in the population activity of ARN KISS neurons were recorded by GCaMP fiber photometry before and following CNO injections in freely behaving mice. b , c Representative 24-h GCaMP photometry traces from the same female mouse injected s.c. with vehicle (VEH, saline) ( b ) and 3 mg/kg CNO ( c ). The shaded area indicates the period of lights off. Peaks represent ARN KISS neuron population SEs. d – f Bar graphs showing the mean + SEM number of SEs during the 4-h baseline and over the first and second 4-h time bins following VEH and different doses of CNO injections s.c. in ( d ) non-hM3Dq-expressing control females ( N = 6), ( e ) hM3Dq-expressing females ( N = 7, * p = 0.039, ** p = 0.007, ** p < 0.0001), and ( f ) male mice ( N = 7, * p = 0.041). g Schematic showing the method for calculating SE onset times and intervals. h – j Mean + SEM graphs showing the onset times of SEs following CNO injection in ( h ) control females ( N = 6), ( i ) hM3Dq-expressing females ( N = 7, *** p = 0.0008), and ( j ) males ( N = 7, ** p = 0.0026). One-way repeated measure ANOVA with Holm-Sidak post hoc comparison ( e ), and two-tailed paired t-tests, adjusted p values with Holm–Sidak method ( d , f , h , I , j ). Parts of this figure were drawn using images from Wikimedia, and SciDraw (licensed under Creative Commons 4.0 and 1.0 licenses, respectively).

Article Snippet: To selectively activate or silence brainstem NA neurons projecting to ARN KISS neurons, mice were injected with FLP-dependent retrograde viral constructs encoding the excitatory (AAVrg-hSyn-fDIO-hM3DGq-mCherry, 1.8 ×10 13 GC/mL, #154868, Addgene, USA), or inhibitory (AAVrg-hSyn-FLEx-FRT-hM4DGi-mCherry, 3.3 ×10 13 GC/mL, #161576 VectorBuilder, USA) DREADD receptors into the middle/caudal ARN (2.0 mm posterior to Bregma, 0.35 mm lateral to midline, 5.93 mm deep from brain surface) at a rate of 100 nL/min for 10 min with a custom-built bilateral Hamilton syringe apparatus.

Techniques: Injection, Activity Assay, Saline, Expressing, Control, Comparison, Two Tailed Test

Journal: Nature Communications

Article Title: Brainstem noradrenergic modulation of the kisspeptin neuron GnRH pulse generator in mice

doi: 10.1038/s41467-025-60837-8

Figure Lengend Snippet: a Cell-attached voltage-clamp trace of a patched hM4Di-expressing LC-NA neuron from a male mouse showing suppressed firing rate after repeated bath applications of 20 μM CNO. b , c Representative GCaMP fiber photometry traces demonstrating the acute excitatory effect of CNO chemogenetic inhibition of ARN-projecting NA neurons compared to vehicle (VEH, saline) in the same mouse. The shaded area indicates the period of lights off. d , e Bar graphs showing the mean + SEM number of ARN KISS neuron SEs in ( d ) hM4Di-expressing diestrous female ( N = 9) and ( e ) male ( N = 7) mice across 4-h time bins before and after s.c. injection of VEH and different doses of CNO. f , g Bar graphs showing the mean + SEM. onset times of SEs following s.c. injection of CNO and VEH in the hM4Di-expresing ( f ) female ( N = 9, * p = 0.0460) and ( g ) male ( N = 7) mice. One-way repeated measure ANOVA with Holm-Sidak post hoc comparison ( d ), and two-tailed paired t -tests, adjusted p values with Holm–Sidak method ( e , f , g ).

Article Snippet: To selectively activate or silence brainstem NA neurons projecting to ARN KISS neurons, mice were injected with FLP-dependent retrograde viral constructs encoding the excitatory (AAVrg-hSyn-fDIO-hM3DGq-mCherry, 1.8 ×10 13 GC/mL, #154868, Addgene, USA), or inhibitory (AAVrg-hSyn-FLEx-FRT-hM4DGi-mCherry, 3.3 ×10 13 GC/mL, #161576 VectorBuilder, USA) DREADD receptors into the middle/caudal ARN (2.0 mm posterior to Bregma, 0.35 mm lateral to midline, 5.93 mm deep from brain surface) at a rate of 100 nL/min for 10 min with a custom-built bilateral Hamilton syringe apparatus.

Techniques: Expressing, Inhibition, Saline, Injection, Comparison, Two Tailed Test