The CXCR3 chemokine receptor has been linked to autoimmune and inflammatory disease, allograft rejection and ischemic nephropathy. CXCR3 is expressed on endothelial- and smooth muscle cells. Although a recent study has posited that antagonizing of CXCR3 function may reduce atherosclerosis, the role of CXCR3 in controlling physiological vascular functions remains unclear. This study demonstrates that disruption of CXCR3 leads to elevated mean arterial pressures (MAP) in anesthetized and conscious mice, respectively. Stimulation of isolated resistance vessels with various vasoconstrictors showed increased contractibility in CXCR3-/- mice in response to angiotensin II (Ang II) and a decreased vasodilatation in response to acetylcholine (ACh). The increased contractibility was related to higher Ang II type 1 receptor (AT1R) expression, while the decreased vasodilatation was related to lower M3-ACh-receptor expression in the mesenteric arteries of CXCR3-/- mice compared to wild-type mice. The vasodilatatory response to ACh could be antagonized by the non-selective ACh-receptor antagonist atropine and the selective M3-receptor antagonist 4-DAMP, but not for M1-, M2- and M4-receptor antagonists. Additionally, the transcription factors SP-1 and EGR-1 bound to the murine AT1R promoter region and increased SP-1 expression in CXCR3-/- mice indicates an imbalanced SP-1 and EGR-1 complex formation which causes increased AT1R expression and hypertension. The data indicate that CXCR3-receptor is important in vascular contractility and hypertension, possibly through upregulated AT1R expression.