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To examine the electrophysiological properties of DmCa\textsubscript{v}3, we injected cRNAs made from the DmCa\textsubscript{v}3 cDNA template into \emph{Xenopus} oocytes.
From 4 days after cRNA injection, DmCa\textsubscript{v}3 was expressed as measured by robust inward currents in 10 mM Ba\textsuperscript{2+} as a charge carrier.
To directly compare the biophysical properties of DmCa\textsubscript{v}3 and mammalian T-type Ca\textsuperscript{2+} channel homolog under the same conditions, we expressed rat Ca\textsubscript{v}3.1 subunit of which biophysical properties were previously reported in the expression system\cite{9495342}.
Compared to Ca\textsubscript{v}3.1, DmCa\textsubscript{v}3 had a similar low-voltage threshold (around
\minus \textminus 60 mV) for activation, although the averaged value is slightly lower by 3 ~ 4 mV.
The current traces of DmCa\textsubscript{v}3 and Ca\textsubscript{v}3.1 were activated and then inactivated during serial step pulses from a holding potential of
-90 \textminus90 mV, producing transient current kinetics with the inactivation kinetics of DmCa\textsubscript{v}3 currents being likely to be slightly slower than those of Ca\textsubscript{v}3.1 currents.
The activation and inactivation kinetics of currents through DmCa\textsubscript{v}3 was accelerated as the higher step pulses were applied, producing a criss-crossing pattern (Fig. 1a), a typical T-type Ca\textsuperscript{2+} channel kinetics.
Analysis of current-voltage (I-V) relationships showed that V\textsubscript{50,act} for half-maximal activation and slope factor (k) of DmCa\textsubscript{v}3 channel are
-43.32 \textminus43.32 $\pm$ 1.38 mV and 7.74 $\pm$ 1.33, while those of Ca\textsubscript{v}3.1 are
-38.92 \textminus38.92 $\pm$ 0.99 and 6.35 $\pm$ 0.94 (Fig. 1a).
These results suggest that DmCa\textsubscript{v}3 can be activated more negative potential than Ca\textsubscript{v}3.1 by 4.4 mV.
Taken together, the biophysical properties of DmCa\textsubscript{v}3 including activation threshold of about
-60 \textminus60 mV, formation of maximal current amplitude at
-20 \textminus20 mV, transient current kinetics, a criss-crossing pattern by currents evoked by a voltage protocol for I-V are very similar to the hallmark properties of native T-type Ca\textsuperscript{2+} channels as well as cloned channels\cite{9495342, 6087159, 9670923, 10066244}.
The activation curves obtained from fitting chord conductance with a Boltzmann equation showed that the potentials (V\textsubscript{50,act}) for half-maximal activation of DmCa\textsubscript{v}3 and Ca\textsubscript{v}3.1 are
-43.32 \textminus43.32 $\pm$ 1.58 and
-38.92 \textminus38.92 $\pm$ 1.15 mV, respectively, indicating that DmCa\textsubscript{v}3 channel activated at 4.4 mV lower test potentials than Ca\textsubscript{v}3.1 (P $<$ 0.05, Student's t-test, n=11 -- 14) (Fig. 1b and Table 1).
In steady-state inactivation, the potentials (V\textsubscript{50,inact}) of 50 \% channel availability for DmCav3 and Cav3.1 are estimated to be -58.04 $\pm$ 0.71 and -61.31 $\pm$ 0.70 mV (P $<$ 0.05, Student’s t-test, n=5 -- 15), indicating that the V\textsubscript{50,inact} of DmCa\textsubscript{v}3 is 3.3 mV more positive than that of Ca\textsubscript{v}3.1 (Fig. 1b and Table 1).
In regard to window current typically designated by the portion overlapped in the steady-state inactivation and activation curves, the window region for DmCa\textsubscript{v}3 is significantly larger than that for Ca\textsubscript{v}3.1, implying that DmCa\textsubscript{v}3 is capable of persistently evoking higher channel activity over relevant voltage range than Ca\textsubscript{v}3.1.
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