Altered Ca²⁺ handling has both immediate physiological effects and long-term genomic effects on vascular smooth muscle function. Previously we have shown that Ca²⁺ entry through voltage-dependent Ca²⁺ channels (VDCCs) or store-operated Ca²⁺ channels (SOCCs) results in phosphorylation of the Ca²⁺/cAMP response element binding protein (CREB) in cerebral arteries. Here, gene transcription profiles resulting from these two Ca²⁺ entry pathways were determined in human cerebrovascular smooth muscle cell cultures using oligonucleotide array analysis. Results were confirmed and expanded using quantitative RT-PCR, Western blot, and immunofluorescence. A distinct, yet overlapping, set of CRE-regulated genes was induced by VDCC activation using K⁺ membrane depolarization versus SOCC activation by thapsigargin. Membrane depolarization selectively induced a sustained increase in early growth response-1 (Egr-1) mRNA and protein that were inhibited by the VDCC blocker, nimodipine and the SOCC inhibitor, 2-APB. Thapsigargin selectively induced a sustained increase in MAP kinase phosphatase-1 (MKP-1) mRNA and protein, and these effects were decreased by 2-APB, but not by nimodipine. The physiologic agonist angiotensin II also stimulated expression of Egr-1 and MKP-1. Co-administration of 2-APB prevented expression of both Egr-1 and MKP-1, whereas nimodipine blocked only Egr-1 expression. Both thapsigargin and angiotensin II induced phosphorylation of ERK that was sensitive to 2-APB and was selectively required for CREB phosphorylation. Our findings thus indicate that Ca²⁺ entry through VDCCs and SOCCs can differentially regulate CRE-containing genes in vascular smooth muscle and also imply that agonist-induced signals involved in the modulation of gene transcription can be controlled by multiple sources of Ca²⁺.