1. alpha(1)-Adrenoceptor (AR) subtypes in mouse carotid arteries were characterised using a combination of agonist/antagonist pharmacology and knockout (KO) mice. 2. Phenylephrine (PE) was most potent in the alpha(1B)-KO (pEC(50)=6.9+/-0.2) followed by control (pEC(50)=6.3+/-0.06) and alpha(1D)-KO (pEC(50)=5.5+/-0.07). Both N-[5-(4,5-dihydro-1H-imidazol-2yl)-2-hydroxy-5,6,7,8-tetrahydronaphthalen-1-yl] methanesulphonamide hydrobromide (A-61603) and 5-hydroxytryptamine (5-HT) were more potent in the alpha(1D)-KO (pEC(50)=7.4+/-0.27 and 7.4+/-0.05, respectively) than the control (pEC(50)=6.9+/-0.09 and 6.9+/-0.08, respectively) and equipotent with the control in the alpha(1B)-KO (pEC(50)=6.7+/-0.07 and 6.8+/-0.04). Maximum responses to PE and A-61603 were reduced in the alpha(1D)-KO compared to control; there was no difference in maximum responses to 5-HT. 3. In control arteries, prazosin and 5-methylurapidil acted competitively with pA(2) of 9.6 and 7.5, respectively. BMY7378 produced antagonism only at the highest concentration used (100 nM; pK(B) 8.3). 4. Prazosin, 5-methylurapidil and BMY7378 acted competitively in alpha(1B)-KO carotid arteries with pA(2) of 10.3, 7.6 and 9.6, respectively. 5. In the alpha(1D)-KO, against PE, 5-methylurapidil produced a pA(2) of 8.1. pK(B) values were calculated for prazosin (10.6) and BMY7378 (7.0). Against A-61603, 5-methylurapidil had a pA(2) of 8.5, prazosin 8.6, while BMY7378 had no effect. 6. In conclusion, the alpha(1B)-KO mediates contraction solely through alpha(1D)-ARs and the alpha(1D)-KO through alpha(1A)-ARs. Extrapolating back to the control from the knockout data suggests that all three subtypes could be involved in the responses, but we propose that the alpha(1D)-AR causes the contractile response and that the role of the alpha(1B)-AR is mainly regulatory.