The patch-clamp method which was rewarded with the Nobel Prize in 1991 is a well-established and indispensable method to study ion channels in living cells and to biophysically characterize non-voltage-gated ion channels that comprise about 70% of all ion channels in the human genome. To investigate the biophysical properties of non-voltage-gated ion channels, whole-cell measurements with application of continuous voltage-ramps are routinely conducted to obtain current-voltage (IV) relationships. However, adequate tools for detailed and quantitative analysis of IV curves are still missing. We use the example of the transient receptor potential classical (TRPC) channel family to elucidate whether the normalized slope conductance (NSC) is an appropriate tool for a reliable discrimination of the IV curves of diverse TRPC channels that differ in their individual curve progression. We provide a robust calculation method for the NSC, and by applying this method we find that TRPC channel activators and modulators can evoke different NSC progressions independent from their expression levels which is pointing to distinguishable active channel states. TRPC6 mutations of patients suffering from focal segmental glomerulosclerosis (FSGS) resulted in distinct NSC progressions suggesting that the NSC is suitable to investigate structure-function relations and might help unravel the unknown pathomechanisms leading to FSGS. Altogether, the NCS represents an effective algorithm for extended biophysical characterization of non-voltage-gated ion channels.
- Hermann, C.
- Treder, A.
- Näher, M.
- Geiseler, R.
- Gudermann, T.
- Mederos, Y. Schnitzler M.
- Storch, U.
