Large conductance cation channels (LCC-channels) are abundantly expressed in the outer and inner nuclear membranes, but a specific antagonist of these channels remains to be identified. Given that many previously discovered modulators of these channels include N-cholinergic receptor agonists and antagonists, this study aimed to assess the blocking effects of M-cholinergic modulators, such as pilocarpine and platyphylline, on LCC-channels. The experiments were performed on the nuclei of cerebellar Purkinje neurons of Wistar rats. The electrophysiological activity of LCC-channels was evaluated based on the currents passing through the channels, which were recorded with the patch-clamp technique in nucleus-attached configuration and voltage-clamp mode. We found that pilocarpine (1 mmol/l) applied to the bath does not affect the openstate probability (Po) of LCC-channels, or the amplitude of the currents through them. In contrast, when applied via the patch pipette, pilocarpine, in addition to reducing Po by 68.3% at -40 mV, decreases the amplitude of the ion currents at +40 mV by 13.9%. Platyphylline (1 mmol/l), applied to the bath solution, decreases both the amplitude of the currents (by 16.9% at -60 mV) and the Po values (by 52.6% at -40 mV) of LCC-channels. However, when added to the patch pipette solution, this substance causes an increase in the amplitude of LCC-channels-mediated currents at negative applied potentials (by up to 20.9% at -40 mV), contrary to the impact observed for in-bath application. Moreover, platyphylline decreases Po of LCC-channels at negative applied potentials (by up to 54.8% at -60 mV), and the amplitude of the currents at positive ones (by up to 22.2% at +40 mV). For both substances, variability in the effects depending on the application configuration may indicate distinct differences in the LCC-channels domains involved in interactions