Amiloride Sensitive Current in hDCT and M1 cells.

Abstract

Introduction: The establishment of a human distal convoluted tubule (hDCT) cell line would significantly advance renal medicine research, as no such human DCT cell line currently exists. This study aims to verify a key component in the development of this cell line by detecting the presence of an amiloride-sensitive sodium current. Methods: The role of epithelial sodium channels (ENaC) in regulating sodium currents was investigated in mouse cortical collecting duct (M1) cells using the whole-cell patch-clamp technique. Results: The data collected from both cell lines demonstrated limited consistency. In the M1 cell line, one out of four cells showed a detectable amiloride-sensitive current after treatment with 10 μM forskolin; however, the reliability of this result is low. A similar pattern was observed in hDCT cells, but no amiloride-sensitive current was conclusively confirmed. In both the M1 and hDCT3 cell lines, the vehicle control exhibited a general trend of current inhibition. Conclusion: The presence of an amiloride-sensitive sodium current could not be conclusively confirmed in either cell line. Further studies, incorporating modifications to the current protocol, may be necessary to achieve more definitive results.