In the characterization of cocaine addiction, various of its rewarding and reinforcing properties have been studied. Recently, in-vivo fiber photometry (FP) monitoring of calcium in the nucleus accumbens (NAc) and prefrontal cortex (PFC) have provide relevant information in the understanding of acute cocaine effects. In addition, another finding have reported the potential effect of the M1 receptor-selective agonist VU0364572 on the cessation of cocaine-seeking behavior. Here, we hypothesized that VU0364572 has some potential neuromodulator properties over the in-vivo dynamics of calcium and glutamate, in response to sub-chronic cocaine exposure (SCE), in the NAc and PFC of female and male mice.Fiber photometry recordings were performed in C57Bl/6J mice between 8 to 10 weeks old, expressing the encoded virus biosensors pGP-AAV-syn-jGCaMP8m-WPRE and AAV2.9-hSyniGluSnFR3-857.GPl. Animals were classified in groups regarding the brain region and sex. Experimental plan: 1st, Saline exposure (SE) 2nd, SCE: cocaine (30mg/kg) for 5 days. After two weeks, 3rd: VU0364572 (3.2 mg/kg). 4th: VU0364572 plus SCE. Finally, histological characterization to check virus expression and fiber tip location.We have characterized the glutamate dynamics in the two brain regions and identified that cocaine reduced the number and amplitude of calcium peaks in the two brain regions, without showing sex differences. These responses were modulated by the pre-exposure to VU0364572 after 2 weeks of withdrawal. These findings suggest that stimulation of the M1 receptor has the ability to regulate the response to cocaine in these two brain regions; Contributing to the understanding of the potential therapeutic mechanisms of this compound.