In the pancreas, the role of the small-conductance, calcium-activated SK channels remains controversial. Here, we show that three SK subtypes are expressed in the rat insulinoma cells. Our findings demonstrate that rat SK1 (rSK1) channels ensure appropriate insulin secretion by establishing the cell’s negative resting membrane potential and shortening the duration of the action potential. We also found that the depletion of rSK1 transcripts generated a condition in which beta cells constitutively secrete insulin, even in the absence of a stimulating molecule (such as glucose). Together, these results implicate SK1 subunits as key regulators of excitability and endocrine function in beta cells.