<p>Ca²⁺ signaling remains essential for excitatory cells’ functioning, but it is still unknown what channels are responsible for the countercurrent of positive ions that accompanies Ca²⁺ release from intracellular stores. Previously, it was suggested that nuclear LCC-channels (Large Conductance Cation channels) may be involved in this process. Considering the results regarding the inhibiting properties of N‑choline receptor modulators on LCC-channels, in this research, we focused on studying the effects of atropine to assess whether it can be used as a&#xa0;blocker of these channels, enabling further evaluation of their role in Ca²⁺ release.</p><p>Electrophysiological properties of LCC-channels were investigated using the patch-clamp technique in voltage clamp mode and nucleus-attached configuration. The amplitude of the currents for a&#xa0;single-channel opening event and the probability of the channels residing in an open state were studied under control conditions and with the solution of atropine applied in two modes: in a&#xa0;bath with a&#xa0;sample, and into a&#xa0;patch pipette.</p><p>We found that atropine effectively decreases the amplitude of the currents through LCC-channels when applied in either mode. When applied to a&#xa0;bath, it has an inhibiting effect only at the negative membrane potentials (−40 mV and −60 mV), while causing an amplitude decrease at both positive (+40 mV and +60 mV) and negative (−60 mV) membrane potentials being applied via a&#xa0;patch pipette. It was also established that the probability of LCC-channels to reside in an open state (P_o) is lowered by atropine applied to a&#xa0;bath at a&#xa0;negative membrane potential of −40 mV, with a&#xa0;similar tendency observed at −60 mV. In the meantime, no changes in P_o were registered when atropine was applied via a&#xa0;patch pipette.</p>