The smooth muscle cell is the principal component responsible for involuntary control of visceral organs, including vascular tonicity, secretion, and sphincter regulation. It is known that the neurotransmitters released from nerve endings increase the intracellular Ca2+ level in smooth muscle cells followed by muscle contraction. We herein report that femtosecond laser pulses focused on the diffraction-limited volume can induce intracellular Ca2+ increases in the irradiated smooth muscle cell without neurotransmitters, and locally increased intracellular Ca2+ levels are amplified by calcium-induced calcium-releasing mechanisms through the ryanodine receptor, a Ca2+ channel of the endoplasmic reticulum. The laser-induced Ca2+ increases propagate to adjacent cells through gap junctions. Thus, ultrashort-pulsed lasers can induce smooth muscle contraction by controlling Ca2+, even with optical stimulation of the diffraction-limited volume.