Eptors inhibit NOS activity. Certainly, NOS activity is enhanced in the cortex of CB1 KO mice (Kim et al., 2006). Additionally, anandamide inhibited anxiousness and panic-like behaviors induced by administration of an NO donor in to the dorsal periaqueductal gray matter (Lisboa and Guimaraes, 2012; Lisboa et al., 2013). Based on these pieces of evidence, the present study investigated the doable involvement of the nitrergic technique in behavioral adjustments stress-related behavioral alterations by testing the hypothesis that: 1) 7-nitroindazole (7-NI), a preferential nNOS inhibitor, would attenuate CFC in WT mice, two) iNOS KO mice would show elevated CFC, three) the enhanced CFC observed in iNOS KO mice would be related to elevated NOS activity and NOS mRNA expression in the MPFC and HIP, and four) the behavioral adjustments observed in iNOS KO mice will be attenuated by 7-NI. We further investigated the involvement of the cannabinoid technique within the modulation of CFC by testing the hypothesis that facilitation of ECB signaling and CB1 antagonism would attenuate and improve CFC, respectively. Moreover, to confirm a possible interaction among cannabinoids and NO, we tested the hypothesis that cannabinoid drugs would modulate CFC and that iNOS KO mice would present modifications in mRNA expression of genes connected towards the ECB system within the MPFC and HIP.MethodsAnimalsAll experiments have been performed applying male C57BL/6J and iNOS KO (C57BL/6J background) mice (82 weeks old). Breeding homozygous pairs of mice with targeted deletion with the iNOS gene had been obtained from Jackson Laboratories (no. 002609, Bar Harbor, ME) and maintained in our local animal farm facility. The animals have been housed in groups of five animals per cage within a temperature-controlled area (24 1oC) below regular laboratory situations (12 h light/12 h dark, lights on at 6:30 am) with food and water accessible ad libitum until they had reached the appropriate age for the experimental procedures.ST6GAL1 Protein manufacturer Animals that received precisely the same treatment had been kept in pairs until the end in the experiments.LIF Protein supplier Procedures had been performed in conformity using the recommendations of the Brazilian Council for the care and use of laboratory animals (COBEA), which comply with international laws and politics, and were approved by our neighborhood ethical committee.PMID:36717102 Experiments were conducted amongst 9:00 am and 3:00 pm.Drugs and Treatment7-NI (15, 30, and 60 mg/kg, Sigma-Aldrich), a preferential nNOS inhibitor, was dissolved in 5 Tween 80 in NaCl 0.9 ; WIN55,212-2 (Win; 0.1, 0.3, and 1.0 mg/kg, Sigma-Aldrich), a nonselective cannabinoid agonist, and AM281 (1, 2, and 4 mg/kg), a potent and selective CB1 antagonist, had been dissolved in 10 dimethylsulfoxide (DMSO) in NaCl 0.9 and administered i.p. to WT mice 30 minutes before the first reexposure to the context chamber (Maren, 1998; Rutkowska et al., 2006; Gilhotra and Dhingra, 2009; Gomes et al.,Lisboa et al. |2011). URB597 (URB; 0.3, 1, and three mg/kg), an inhibitor of the FAAH enzyme that metabolizes the ECB anandamide, was dissolved in ten DMSO in NaCl 0.9 and administered i.p. to WT mice 1 hour just before the first reexposure to the context chamber (Gomes et al., 2011). All drugs had been administered in a fixed volume of ten mL/ kg of body weight. The animals have been also reexposed towards the exact same context 48, 72, and 96 hours immediately after the first chamber exposure. For evaluation of freezing behavior, independent groups of WT and iNOS KO mice received i.p. injections of 7-NI (successful dose, 30 mg/kg) or URB (effective.
