In vitro monitoring of cytochrome oxidase activity in neuronal cells

The purpose of this investigation is to design a system to measure cellular activity in relation to the activity of cytochrome c oxidase (Cox), a key metabolic enzyme found in all cell types. Cox levels in neuronal cells have been found to be high when compared to other cell types. This system is being developed to investigate cell metabolism of neurons growing in a micro-machined silicon glass chamber designed for studying neuronal growth. Cox has also been linked with neurodegenerative disorders such as Alzheimer´s and Parkinson´s disease and may have a role as a marker for such diseases. Here, the authors explore the possibility of using NIRS (Near Infrared Spectroscopy) 845 nm region using FTIR (Fourier Transform Infra-Red) spectroscopy. Changes in the oxidation state of Cox can be seen as changes in absorption at 845 nm. As part of this investigation, quantification and analysis of Cox was also carried out using Western blot protein analysis, immunochemistry and fluorescence microscopy to detect levels in neuroblastoma (NG115-401 L) and primary rat neuronal cells