Abstract
We present the design of an innovative molecular neuroprotective strategy, and provide proof-of-concept for its implementation, relying on the injury-mediated activation of an ectopic gene construct. As oxidative injury leads to the intracellular liberation of zinc, we hypothesize that tapping onto the zinc-activated metal regulatory element (MRE) transcription factor 1 (MTF-1) system to drive expression of the Kv2.1-targeted hepatitis C protein NS5A, will provide neuroprotection by preventing cell death-enabling cellular potassium loss in rat cortical neurons in vitro. Indeed, using biochemical and morphological assays, we demonstrate rapid expression of MRE-driven products in neurons. Further, we report that MRE-driven NS5A expression, induced by a slowly evolving excitotoxic stimulus, functionally blocks injurious, enhanced Kv2.1 potassium whole-cell currents and improves neuronal viability. We suggest this form of "on-demand" neuroprotection could provide the basis for a tenable therapeutic strategy to prevent neuronal cell death in neurodegeneration.
- apoptosis
- electrophysiology
- excitotoxicity
- gene therapy
- Kv channels
- metallothionein
- metals and chelators
- neurodegeneration
- voltage gated channels
- voltage-gated potassium channels
- The American Society for Pharmacology and Experimental Therapeutics