Abstract
Using proteomics, previous work in our laboratory identified five mitochondrial related proteins [citrate synthase (CS), glucose-regulated protein 75 (GRP75), heat shock protein 60 (HSP60), prohibitin (PHB), voltage-dependent anion channel 1 (VDAC1)] to be differentially expressed in primary cortical neuronal cultures following preconditioning treatments [1] [2]. To investigate a protective or damaging role of these five proteins in neurons, we used RNAi constructs to knockdown and adenoviral vectors to over-express the proteins in cortical neuronal cultures prior to exposure to three ischemia-related injury models: excitotoxicity (L-glutamic acid), oxidative stress (hydrogen peroxide) and in vitro ischemia (oxygen-glucose deprivation). We observed that down-regulating these mitochondrial proteins had no effect on neuronal viability, in any injury model. By contrast, over-expression of PHB exacerbated cell death in the hydrogen peroxide and L-glutamic acid injury models. These findings indicate that PHB plays a neurodamaging role following oxidative and excitotoxic stress and suggests that the protein is a potential therapeutic target for the design of drugs to limit neuronal death following cerebral ischemia and other forms of brain injury.
Highlights
Acute brain injury following stroke and other forms of cerebral ischemia have an immerse impact on patients and their families and on the broader community, due to the high social and economic cost
Western blot analysis confirmed that Short Interfering RNA (siRNA) constructs for citrate synthase (CS), glucose-regulated protein 75 (GRP75), heat shock protein 60 (HSP60), PHB and VDAC1decreased the expression of their respective target protein in cortical neuronal cultures (Figure 1)
Cortical neuronal cultures treated with specific siRNA constructs and subjected to in vitro ischemia, hydrogen peroxide and L-glutamic acid exposure, did not result in any statistically significant increase or decrease in neuronal viability (Figure 2)
Summary
Acute brain injury following stroke and other forms of cerebral ischemia have an immerse impact on patients and their families and on the broader community, due to the high social and economic cost. Major risk factors for stroke include hypertension, smoking, lack of physical activity, diabetes and an unhealthy diet [3]. Brain damage following a stroke manifests itself through a range of symptoms including problems with coordination and speech, paralysis in one or more limbs, dizziness, visual defects and reduced alertness or concentration [4]. As neuroprotective agents capable of limiting ischemic brain injury are unavailable, a major objective in stroke research is the identification of potential therapeutic targets for the development of clinically effective neuroprotective drugs. One approach to identify therapeutic targets for neuroprotective drug discovery is to investigate differential protein expression following neuronal preconditioning or the induction of ischemic tolerance. As preconditioning is reliant on new protein synthesis, the identification of proteins involved in this process is a potential target in the development of drugs to inhibit ischemic neuronal death
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
