Male and female PACAP-Cre transgenic mice (n = 15 males, 11 females) underwent the binge-type eating paradigm, and received injections into the PVT of a Cre-dependent excitatory DREADD (n = 14) or control virus (n = 12) during the 3rd or 4th week of the paradigm. Following 6 total weeks of binge-type eating, all mice received intraperitoneal (i.p.) injections of 5 mg/kg clozapine n-oxide (CNO; HelloBio, Princeton, NJ, USA) or saline vehicle (~0.14 mL; Baxter International Inc., Deerfield, IL, USA), 30 minutes prior to their binge eating session, and subsequent intake was recorded. These injections were made within-subject, with 48 hours between injections. One week later, to ensure that Cre was co-expressed with PACAP peptide, and that injection of CNO resulted in excitation of these Cre+ cells, a sub-set of the mice was injected between-subject with CNO or saline and sacrificed 120 minutes later. Immunohistochemistry was then used to detect Cre, PACAP, and c-Fos.
Clozapine n oxide cno
Manufactured by Hello Bio
Sourced in United States
Clozapine n-oxide (CNO) is a chemical compound used as a pharmacological tool in research applications. It is a synthetic molecule that serves as an inert metabolite of the antipsychotic drug clozapine. CNO is commonly used in combination with designer receptors exclusively activated by designer drugs (DREADDs) technology to remotely control the activity of genetically-modified cells in biological research.
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Lab products found in correlation
5 protocols using clozapine n oxide cno
1
PACAP-Cre Mice Binge Eating Paradigm
2
Chronic Morphine Exposure in Mice
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Morphine sulfate was supplied by NIDA Drug Supply (Research Triangle Park, NC) and dissolved into a 0.2% saccharin solution so that mice would consume morphine via drinking water [31 ]. Saccharin was used in the baseline condition and in the control groups for the duration of the studies. Morphine concentration was gradually increased from 0.3 to 0.5 mg/ml to ensure stable drinking throughout the 11 days of chronic exposure. Dosing stayed at 0.5 mg/ml for the remainder of the experiments [32 (link)]. Body weights and drinking volume were assessed every 2–3 days to confirm morphine ingestion, and adequate body weight was maintained. Clozapine N-Oxide (CNO) (HelloBio, cat. no. 1807) was dissolved in saline and administered i.p. at 1 mg/kg.
3
Pharmacological Modulation of BLA during Stress
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Before drug infusion, mice were gently restrained, and dummy cannula were replaced with injection cannula that extended 1 mm from the tip of the guide cannula. For protein kinase A (PKA) inhibition during a stress-inducing session, saline or Rp-cAMP (Santa Cruz Biotechnology, TX, USA) was infused into the lateral ventricle or the BLA using a 10 μL Hamilton syringe under infusion pump control. For chemogenetic inhibition of OFC–BLA transmission, saline or clozapine n-oxide (CNO, Hello Bio, Bristol, UK) were infused into the BLA.
4
Automated Fecal Collection and CNO Administration
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We transferred control and hM4Di-expressing mice to our custom-built feces collectors and allowed them to acclimate to the cages in LD for at least 7 d before beginning fecal pellet collection. We replaced the water bottles on the cages with 1% sucrose water daily for 3 d prior to the start of collection to acclimate mice to the removal and replacement of their water bottles. We then automatically collected fecal pellets for three days. From CT 6 on the second day of collection until CT 6 the following day, we replaced the water bottles on all cages with clozapine-N-oxide (CNO, 1 mg/kg; Hello Bio, Princeton, NJ) in 1% sucrose water (to mask the bitter taste of CNO). This method of continual CNO administration in the drinking water has been previously shown to successfully alter neuronal activity for the duration of CNO administration121 (link),122 (link). Virus expression was similar between mice. Mice consumed between 5 and 8 ml of CNO water, receiving at least 1 mg/kg CNO throughout the day; there was no genotype difference in the amount consumed.
5
Optogenetic Inhibition of SF1 Neurons
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Gi-Designer Receptors Exclusively Activated by Designer Drugs (DREADDs)-AAV (Gi-DREADD), rAAV8-hSyn-DIO-hM4D(Gi)-mCherry (Addgene Inc, Catalog# 44362-AAV8, 3.2 × 1012 VM/mL), was used for inhibition of VMHdm/cSF-1 neuronal activity induced by optogenetics. rAAV8-hSyn-DIO-hM4D(Gi)-mCherry and rAAV5-EF1α-DIO-ChR2(H134R)-EYFP (250 nL each, total 500 nL) were injected into the right side of the VMH of Sf-1-Cre mice as described above. Control for rAAV5-EF1α-DIO-ChR2(H134R)-EYFP was rAAV5-EF1α-DIO-EYFP. The optic cannula was inserted the VMH, PVH/AH or PAG as described above. Clozapine-N-Oxide (CNO, catlog# HB6149, Hello Bio Inc, NJ, USA) was injected intraperitoneally (i.p.) 10 min before optogenetic stimulation begun (2.5 mg/kg per BW). Sterile saline was used as vehicle and control.
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