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PACAP induces increased excitability in D1- and D2-expressing nucleus accumbens medium spiny neurons

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 17, 2025)
Funding
CONAHCyT 1102522
Abstract
One of the main eating disorders associated with overweight and obesity is binge eating disorder. Binge eating is characterized by excessive consumption of high-calorie foods over a short period of time, approximately 2 hours. The nucleus accumbens (NAc) plays a key role in modulating the hedonic value of high-calorie foods, commonly referred to as palatable foods. Specific subregions of the shell portion of the NAc (NAcSh), known as hedonic hot spots, may play an important role in the motivational aspect of food consumption. Previous work has shown that the pituitary adenylate cyclase-activating polypeptide (PACAP) injected into the NAc reduces palatable food intake, suggesting that this peptide could be a potential tool for treating binge eating. However, the mechanisms of action of PACAP on the NAc are poorly understood. Here, we used whole-cell recording and calcium imaging techniques in NAcSh brain slices from D1-Cre and A2A-Cre mice to investigate PACAP modulation of medium spiny neuron (MSN) activity. We found that PACAP administration increased the firing rate of D1- and D2- expressing MSNs. In addition, in a binge-eating mouse model, nasal PACAP reduced binge-eating behavior.
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