Divalent cation permeability and blockade of Ca2+-permeant non-selective cation channels in rat adrenal zona glomerulosa cells

摘要

class="enumerated" style="list-style-type:decimal">The effects of the divalent cations Ca²⁺, Mg²⁺ and Ni²⁺ on unitary Na⁺ currents through receptor-regulated non-selective cation channels were studied in inside-out and cell-attached patches from rat adrenal zona glomerulosa cells.External Ca²⁺ caused a concentration-dependent and voltage-independent inhibition of inward Na⁺ current, exhibiting an IC50 of 1.4 mm. The channel was also Ca²⁺ permeant and external Ca²⁺ shifted the reversal potential as expected for a channel exhibiting a constant Ca²⁺:Na⁺ permeability ratio near to 4.External and internal 2 mm Mg²⁺ caused voltage-dependent inhibition of inward and outward Na⁺ current, respectively. Modelling Mg²⁺ as an impermeant fast open channel blocker indicated that external Mg²⁺ blocked the pore at a single site exhibiting a zero voltage Kd of 5.1 mm for Mg²⁺ and located 19% of the distance through the transmembrane electric field from the external surface. Internal Mg²⁺ blocked the pore at a second site exhibiting a Kd of 1.7 mm for Mg²⁺ and located 36% of the distance through the transmembrane electric field from the cytosolic surface.External Ni²⁺ caused a voltage- and concentration-dependent slow blockade of inward Na⁺ current. Modelling Ni²⁺ as an impermeant slow open channel blocker indicated that Ni²⁺ blocked the pore at a single site exhibiting a Kd of 1.09 mm for Ni²⁺ and located 13.7% of the distance through the transmembrane electric field from the external surface.External 2 mm Mg²⁺ increased the Kd for external Ni²⁺ binding to 1.27 mm, consistent with competition for a single binding site. Changing ionic strength did not substantially affect Ni²⁺ blockade indicating the absence of surface potential under physiological ionic conditions.It is concluded that at least two divalent cation binding sites, separated by a high free energy barrier (the selectivity filter), are located in the pore and contribute to Ca²⁺ selectivity and permeability of the channel.