Activation of Voltage-Gated Calcium Channels (VGCC) by nsEP

Nanosecond pulses may be too brief to cause translocation of the voltage sensor of VGCC and channel opening. Instead, nsEP can electroporate the membrane, causing depolarization due the loss of the resting membrane potential, which will culminate in the VGCC opening.

This project aims to establish if VGCC can be activated by nsEP directly, without the concurrent membrane damage by electroporation.

HEK293 cells do not express endogenous VGCC. We transfect them to express VGCC (CaV1.3, red marker) and test if the VGCC is functional by the entry of Ca2+ (green) upon depolarization in KCl. Next, we compare activation and membrane permeabilization by nsEP in cells which do and do not express functional VGCC.
Olympus IX81 microscope equipped for image recording concurrently with nsEP exposure, drug and solution delivery, and patch-clamp data acquisition