The Effect of Nickel and Manganese on Store-Operated Channels in Canine Pulmonary Arterial Smooth Muscle Cells

Yeom, Jong Hoon; Min, Hyoung Ki; Shim, Jae Hang; Jeon, Woo Jae; Cho, Sang Yoon; Yoon, Woo Jong; Kim, Kyoung Hun

Abstract

BACKGROUND
Pulmonary vasoconstriction greatly contributes to the elevated pulmonary vascular resistance in patients with pulmonary hypertension. A rise in cytosolic Ca2+ concentration ([Ca2+]cyt) in pulmonary arterial smooth muscle cells (PASMCs) is a major trigger for pulmonary vasoconstriction. Presently, little is known about the nature of the store-operated channels (SOCs) in PASMCs, even though store depletion-mediated capacitative Ca2+ entry (CCE) is a critical mechanism for refilling the empty sarcoplasmic reticulum (SR) with Ca2+, and for maintaining a sustained increase in [Ca2+]cyt. The goal of this in vitro study was to investigate the effects of nickel and manganese on store-operated channels in canine PASMCs.
METHODS
Isolated PASMCs were obtained from an enzymatically treated canine pulmonary artery. Currents were recorded at room temperature using the dialyzed whole cell recording technique. The protocol used to deplete the intracellular Ca2+ stores and to monitor the development of the store-operated Ca2+ currents, involved voltage-clamping cells at 0 mV to inactivate any voltage-dependent calcium currents. Crrents were recorded in response to a 200 ms voltage step from 120 to 40 mV in 20 mV increments every 15 seconds.
RESULTS
Simultaneous depletion of intracellular Ca2+ leads to a voltage-independent and linear store-operated Ca2+ current (ISOC) reversal near 0 mV. Nickel and manganese inhibit ISOC.
CONCLUSIONS
In canine PASMCs, the depletion of intracellular Ca2+ stores leads to the activation of ISOC, which is inhibited by nickel and manganese. These metals are non-specific inhibitors of non-selective cation channels. Our results indicate that Ni2+- and Mn2+-sensitive pathways may mediate Ca2+ entry, or that a class of non-selective cation channels may contribute to CCE in canine PASMCs.