Experiment/han 2023 mouse arcuate nucleus
From Miniscope
Mouse arcuate nucleus imaging of kisspeptin neurons
Deep hypothalamic imaging of the kisspeptin pulse generator in freely behaving male mice.
Extracted from full textTranscribed by a curator from the published methods section. Values not stated in the paper are marked as not reported rather than inferred.
Animal
Species
Mouse
Transgenic line
Kiss1-Cre (129S6Sv/Ev x C57BL/6 background)
Sex
Male
Age at surgery
8-14 weeks
Animals
5
Target
Brain region
Coordinates
AP -2.0 from bregma, ML 0.3 from midline; lens 200 um above the injection site
Hemisphere
Not reported
Cell type
Kisspeptin (Kiss1-expressing) neurons
Indicator
Indicator
GCaMP6s
Reports
Calcium
Delivery
AAV + transgenic
Construct
AAV9-CAG-FLEX-GCaMP6s-WPRE-SV40
Titer
1.3 x 10^13 GC/mL
Injection volume
1 uL
Expression time
5 weeks
Optical access
Method
GRIN lens
Lens
500 um diameter, 8.4 mm long (Grintech)
Implant depth
200 um above the injection site
Surgery staging
Single surgery
Implant to baseplating
1 week
Imaging
Scope
Condition
Freely behaving
Frame rate
30 Hz
Sessions
Habituated for at least 3 weeks after baseplating
Analysis
NoRMCorre, CNMF-E, MiniscopeAnalysis (MATLAB)
Cell yield
13-50 neurons per animal
What was hard about this preparation
An 8.4 mm lens at 500 um diameter is among the most demanding implants represented here: the tract passes through the full dorsoventral extent of the brain, and cell yield stays modest (13-50 per animal) even when it works.
Who to ask
Experimenter
Allan E. Herbison
Institution
University of Cambridge
Notes
Long habituation — at least 3 weeks post-baseplate — before recording.