F- (10 mM sodium fluoride plus deferoxamine to chelate contaminating aluminum) causes arterial contractions primarily by activating L-type Ca2+ channels. Results from the present study indicate that, although F(-)-induced contractions could be completely relaxed by washing out the F- with fresh buffer, a long-lasting effect of F- pretreatment was to produce L-type Ca2+ channel desensitization. Pretreatment of arteries for 4 h with F- (followed by washout of F-) resulted in much reduced increases in stress and [Ca2+]i produced by the subsequent addition of 110 mM KCl, such that steady-state values were, respectively, only 9 and 15% of the control values. However, a 4-h F- pretreatment caused a reduction only in the rate of stress development, but not the steady-state level of stress, produced by maximum concentrations of receptor agonists. In tissues that were pretreated with F- and then stimulated with the alpha-adrenoceptor agonist, phenylephrine, steady-state stress was still 104% of the control value, while the increase in [Ca2+]i was only 10% of the control value. F- is known to inhibit protein phosphatases, and similar reductions in the ability of KCl to produce contractions and increase [Ca2+]i were seen after pretreatment with the protein phosphatase inhibitor, okadaic acid. These data suggest that L-type Ca2+ channel desensitization by F- pretreatment was caused by increased protein phosphorylation. In addition, they suggest that much of the contribution made by L-type Ca2+ channels to increase [Ca2+]i during receptor stimulation may not be necessary for the maintenance of maximum stress at steady state.