Paav hsyn hm4d gi mcherry

Manufactured by Addgene

Sourced in United States

PAAV-hSyn-hM4D(Gi)-mCherry is a plasmid construct that contains the human Synapsin 1 (hSyn) promoter, the human M4 muscarinic receptor (hM4D(Gi)) coding sequence, and the mCherry fluorescent protein coding sequence. This plasmid can be used for the expression of the inhibitory DREADD (Designer Receptors Exclusively Activated by Designer Drugs) receptor hM4D(Gi) in neurons.

Automatically generated - may contain errors

Lab products found in correlation

Paav hsyn hm3d gq mcherry, by Addgene (2 mentions)

4 protocols using paav hsyn hm4d gi mcherry

Targeted DREADD Expression in Prefrontal Cortex

Check if the same lab product or an alternative is used in the 5 most similar protocols

For some experiments, an AAV retrograde virus expressing pAAV-hSyn-hM4D(Gi)-mCherry (Addgene #50475) or pAAV-hSyn-hM3D(Gq)-mCherry (Addgene #50474) was injected into either the ipsilateral pons (relative to lambda, 1.00 mm lateral, 0.20 mm posterior, depth of 4.55 mm) or contralateral mPFC (relative to bregma, 0.48 mm lateral, 2.1 mm anterior, depth of 1.6 mm) of 6- to 9-week-old mice to express DREADD (“designer receptor exclusively activated by designer drugs”) receptors selectively in PT (ipsilateral pons injections) or IT (contralateral mPFC injections) neurons in the prefrontal cortex. Under sterile surgical conditions, animals were anesthetized with continuous isoflurane (~2%) and a craniotomy made at the above coordinates. A 33 gauge microsyringe (Hamilton) containing undiluted virus was slowly lowered into place over a period of ~5 minutes. After waiting anther 5 minutes, the virus was injected at a rate of 50 nL per minute for a total volume of 450 nL (pons) or 300 nL (mPFC). Five minutes after the injection was completed, the microsyringe was slowly removed and the wound sutured. Mice were allowed to recover from surgery for at least 21 days before use in experiments.

, & Gulledge A.T. (2023). Cholinergic activation of corticofugal circuits in the adult mouse prefrontal cortex. bioRxiv.

+ Open protocol

+ Expand

Optogenetic and Chemogenetic Manipulations of Cocaine Addiction

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cocaine (15 mg/kg in saline; Macfarlan Smith Ltd, UK), AAV2-hSyn-EYFP (control virus; viral titer ≥ 7 × 10¹² vg/mL; 0.5 μL/loci), adeno-associated virus-2 vectors driven by human alpha-synuclein promoter expressing halorhodopsin (AAV2-hSyn-eNpHR3.0-EYFP, NPHR; viral titer ≥ 7 × 10¹² vg/mL; 0.5 μL/loci) or channelrhodopsin (AAV2-hSyn-ChR2 (E123A)-EYFP, ChR; UNC viral vector core, NC, USA; viral titer ≥ 7 × 10¹² vg/mL; 0.5 μL/loci), adeno-associated virus vectors (pAAV-hSyn-hM4D(Gi)-mCherry, Addgene, Watertown, MA, USA; viral titer ≥ 10¹² vg/mL; 0.5 μL/loci) for inhibitory designer receptors exclusively-activated by designer drugs (DREADDi), and clozapine N-oxide dihydrochoride (CNO, Tocris Bioscience, Bristol, UK) were used.

Jang H.B., Ahn D., Chang S., Kim H.K., Lee B.H., Kim S.C., Steffensen S.C., Bills K.B., Lee H, & Kim H.Y. (2023). Activation of a hypothalamus-habenula circuit by mechanical stimulation inhibits cocaine addiction-like behaviors. Biological Research, 56, 25.

+ Open protocol

+ Expand

DREADD-Expressing Adeno-Associated Viruses

Check if the same lab product or an alternative is used in the 5 most similar protocols

Adeno-associated viruses containing constitutive G_i-coupled (pAAV-hSyn-hM4D(Gi)-mCherry) and G_q-coupled (pAAV-hSyn-hM3D(Gq)-mCherry) DREADDs were obtained from Addgene (viral prep 50,475-AAV8 and 50,474-AAV8 respectively) and had titers of ≥ 7 × 10¹² GC/ml and 2.7^13 GC/ml.

Reich N.C, & Pfeffer L.M. (1990). Evidence for involvement of protein kinase C in the cellular response to interferon alpha.. Proceedings of the National Academy of Sciences of the United States of America, 87(22), 8761-8765.

+ Open protocol

+ Expand

Chemogenetic Silencing of NAc-projecting vHPC Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols

The chemogenetic inhibitory approach was used to silence NAc-projecting vHPC glutamatergic neurons and determine the effects of this circuit on FST performance. Six weeks before ending HFD, the rAAV expressing the engineered human M4 muscarinic receptor couples with Gi protein (hM4D_Gi) and mCherry (titer ≥ 7 × 10¹² vg/mL; pAAV-hSyn-hM4D_Gi-mCherry, Cat# 50475-AAVrg, Addgene) was bilaterally infused into the NAc of mice. On the experiment day, CNO (Cat# 4936, Tocris Bioscience, Bristol, UK) was bilaterally infused into the vHPC (2 µg/µL dissolved in saline, 1 µL/side) of free-moving mice, 30 min before the depression-like behavioral test, to transiently inhibit neuronal activity [31 (link)]. Mice were sacrificed 120 min after the completion of CNO infusions. Those mice that received bilateral infusions of an equal volume of saline to the vHPC served as control. Please see Supplementary 1 for detailed protocols of rAAV production and infusion.

Tsai S.F., Hsu P.L., Chen Y.W., Hossain M.S., Chen P.C., Tzeng S.F., Chen P.S, & Kuo Y.M. (2022). High-fat diet induces depression-like phenotype via astrocyte-mediated hyperactivation of ventral hippocampal glutamatergic afferents to the nucleus accumbens. Molecular Psychiatry, 27(11), 4372-4384.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!