Objective: Several members of the PKD gene family (PKD2, PKDL and PKD2L2) are expressed in the heart. Polycystin-2 and its homologues, which are encoded by these genes, have recently been shown to form Ca²⁺-regulated nonselective cation channels in heterologous expression systems. Previously, large conductance nonselective cation channels (LCC) have been described in cardiomyocytes, however, their molecular identity remained obscure. We therefore examined whether LCCs may be formed by polycystins. Methods: Myocytes isolated from the rat left ventricle were investigated by the whole-cell patch-clamp technique and single-cell RT-PCR. Results: Application of 10 mM caffeine to the bath solution to increase the intracellular Ca²⁺ concentration led to activation of LCC in 56% of the myocytes investigated (total n = 651), in ∼10%, more than three LCCs were detected. The single channel conductance was ∼300 pS for monovalent cations and the channel was relatively nonselective for the monovalent cations Na⁺, K⁺, Li⁺, and Cs⁺ and also permeable for the divalent cations Ca²⁺ and Ba²⁺, but impermeable for NMDG⁺ and Cl⁻. Amiloride (IC₅₀=131±1.1 μM) and millimolar concentrations of the trivalent cations Gd³⁺ and La³⁺ inhibited the LCC. Single-cell RT-PCR analysis revealed that mRNA of PKD2 and PKD2L2, but not PKDL or PKD1 are expressed in individual rat left ventricular myocytes. Conclusion: The characteristics of LCC shown in the present study are nearly identical to those observed for polycystin-2 and its homologues suggesting that polycystin-2 or polycystin-2L2 underlie LCC in ventricular myocytes.