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Syn.Flex.GCaMP6f.WPRE.SV40 (AAV9) in nucleus accumbens shell (NAcSh) brain slices from D1-Cre and A2A-Cre mice

Addgene #100833-AAV9

Data Submitted By
Sonia Ortega-Tinoco
Lab Name
Salvador Hernández-López, Departamento de Fisiología, Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM)
Submission Date
September 08, 2025
Publication Date
October 17, 2025 (modified October 20, 2025)
DOI
https://doi.org/10.57733/addgene.a8yf1k
Funding
CONAHCyT 1102522
Abstract
Pituitary adenylate cyclase-activating polypeptide (PACAP) modulation of medium spiny neuron (MSN) activity was investigated through whole-cell recording and calcium imaging of nucleus accumbens shell (NAcSh) brain slices from D1-Cre and A2A-Cre mice. D1 and D2 medium spiny neurons expressing GCaMP6f were labeled via transduction with 100833-AAV9: pAAV.Syn.Flex.GCaMP6f.WPRE.SV40.
Citation
Syn.Flex.GCaMP6f.WPRE.SV40 (AAV9) in nucleus accumbens shell (NAcSh) brain slices from D1-Cre and A2A-Cre mice. Ortega-Tinoco S 2025. Addgene Report, https://doi.org/10.57733/addgene.a8yf1k
Caption: Whole-cell recording and calcium imaging in NAcSh coronal brain slices from D1-Cre mice at 15 μm. D1 medium spiny neurons expressing GCaMP6f are labeled via transduction with 100833-AAV9: pAAV.Syn.Flex.GCaMP6f.WPRE.SV40.
Image attribution: Sonia Ortega-Tinoco
Caption: Whole-cell recording and calcium imaging in NAcSh coronal brain slices from D1-Cre mice at 20 μm. D1 medium spiny neurons expressing GCaMP6f are labeled via transduction with 100833-AAV9: pAAV.Syn.Flex.GCaMP6f.WPRE.SV40.
Image attribution: Sonia Ortega-Tinoco
Caption: Whole-cell recording and calcium imaging in NAcSh coronal brain slices from A2A-Cre (D2 expressing) mice. D2 medium spiny neurons expressing GCaMP6f are labeled via transduction with 100833-AAV9: pAAV.Syn.Flex.GCaMP6f.WPRE.SV40.
Image attribution: Sonia Ortega-Tinoco

This report is made available under the Creative Commons Attribution 4.0 International License.

Vector Used
pAAV.Syn.Flex.GCaMP6f.WPRE.SV40
Addgene #100833-AAV9

Virus & Injection

Virus

Virus Name
pAAV.Syn.Flex.GCaMP6f.WPRE.SV40
Serotype
AAV9
Source
Addgene
Catalog Number
100833-AAV9
Promoter
Synapsin
Cargo Type
Biosensor
Injection Titer / Dose
Bilateral injections of 0.6 μL of 100833-AAV9: pAAV.Syn.Flex.GCaMP6f.WPRE.SV40 were administered into the nucleus accumbens shell (NAcSh) using a 30G dental needle attached to a microinjection system, at a rate of 0.1 μL/min.
Injection Volume
0.6 μL
Injection Rate
0.1 μL/min
Injector Material
30G dental needle
Injection Site / Route
Nucleus accumbens shell (NAcSh) at stereotaxic coordinates relative to bregma: AP +1.1 mm, ML ±1.1 mm, DV -5.2 mm.
View in Addgene Catalog

Other Details

Species
Mouse
Strain or Cell Line
C57BL/6J, D1-Cre mice (Drd1a-Cre), and D2-Cre mice (Adora2a-Cre)
Source
D1-Cre mice: Jax stock #030329; D2-Cre mice: MMRRC_031168-UCD
Age at Injection
30 days
Sex
Male
Time After Injection
18–21 days
Detection Method
Whole-field fluorescence
Assay & Results
The image field was visualized under a microscope and the field size was 280 ×260 μm size. The tissue was stimulated with light pulses (488 nm wavelength, 15–25 ms exposure time) using a LED lamp (Lambda HpX L5, Sutter Instruments) connected to the microscope via fiber optics. Image sequences were acquired at a frequency of 4 frames/s (250 ms/frame) by using a digital camera (Cool SNAP MYO, Photometrics)
Tips & Comments
Please see the original publication for more details: Ortega-Tinoco, S., Padilla-Orozco, M., Hernández-Vázquez, F., Garduño, J., Mondragón-García, A., Ramírez-Sánchez, E., Bargas, J., & Hernández-López, S. (2025). PACAP induces increased excitability in D1- and D2-expressing nucleus accumbens medium spiny neurons. Brain research bulletin, 224, 111323. https://doi.org/10.1016/j.brainresbull.2025.111323