Fig. 5

Immunohistochemical analysis of CB1R and CB2R. A Immunohistochemistry of naïve male and female cortex. Staining for CB1R, CB2R, Iba1, GFAP, and Nissl was performed. Original magnification ×63. B Immunohistochemistry of naïve male and female PAG. Staining for CB1R, CB2R, Iba1, GFAP, and Nissl was performed. Original magnification ×63. Scale on lower left image applies to all frames. C Relative immunoreactivity of CB1R staining revealed significantly greater staining for CB1R in female cortex and lPAG vs males for both regions (****p < 0.0001 female cortex vs male cortex, ***p = 0.001 female lPAG vs male lPAG). D Measurement of co-efficient of colocalization for CB1R with Iba1, Nissl, and GFAP. CB1R colocalization with Nissl was significantly higher in female cortex vs male cortex (***p = 0.008) and female PAG vs male PAG (**p = 0.01). E CB2R immunoreactivity was significantly higher in female lPAG vs male lPAG (**p = 0.0053). F Measurement of co-efficient of colocalization for CB2R with IBa1, Nissl, and GFAP. CB2R colocalization with Nissl in the male cortex was significantly greater than in female cortex (***p = 0.0178). All calculations for co-efficient of colocalization were performed with respect to the relevant enzyme. n = 3/sex for each analysis