The activity of persistent Ca²⁺ sparklets, which are characterized by longer and more frequent channel open events than low-activity sparklets, contributes substantially to steady state Ca²⁺ entry under physiological conditions. Here, we addressed two questions related to the regulation of Ca²⁺ sparklets by PKCα and c-Src, both of which increase whole-cell Ca_v1.2 current: 1) Does c-Src activation enhance persistent Ca²⁺ sparklet activity? 2) Does PKCα activate c-Src to produce persistent Ca²⁺ sparklets? Using TIRF microscopy, Ca²⁺ sparklets were recorded from voltage-clamped tsA-201 cells co-expressing wild type (WT) or mutant Ca_v1.2c (the neuronal isoform of Ca_v1.2) constructs ± active or inactive PKCα/c-Src. Cells expressing Ca_v1.2c exhibited both low-activity and persistent Ca²⁺ sparklets. Persistent Ca²⁺ sparklet activity was significantly reduced by acute application of the c-Src inhibitor PP2 or co-expression of kinase-dead c-Src. Ca_v1.2c constructs mutated at one of the two C-terminal residues (Y²¹²²F, Y²¹³⁹F) were used to test the effect of blocking putative phosphorylation sites for c-Src. Expression of Y²¹²²F but not Y²¹³⁹F Ca_v1.2c abrogated the potentiating effect of c-Src on Ca²⁺ sparklet activity. We could not detect a significant change in persistent Ca²⁺ sparklet activity or density in cells co-expressing Ca_v1.2c + PKCα, regardless of whether WT or Y2122F Ca_v1.2c was used, or after PP2 application, suggesting that PKCα does not act upstream of c-Src to produce persistent Ca²⁺sparklets. However, our results indicate that persistent Ca²⁺ sparklet activity is promoted by the action of c-Src on residue Y²¹²² of the Ca_v1.2c C-terminus.