Abstract
Several recent studies on the function of neuroglobin (Ngb), a hemoprotein predominantly expressed in the brain, point toward a neuro-protective role during hypoxic-ischemic injuries. The exact mechanism by which Ngb protects the cell against H2O2-induced cell death remains to be elucidated. Hence, new tools need to be developed in order to study the proteinin vivoor under physiological conditions. In this summary of our work, we demonstrate how resonance Raman spectroscopy, optical tweezers and microfluidic systems were combined to mimicin vivoconditions in anin vitromilieu. The setup has been tested on several globin-containing cells: hemoglobin (Hb) within single red blood cells (RBCs), a nerve globin present in the nerve cord of the annelidAphrodite aculeata(A. aculeata), and wild-type (wt) human neuroglobin (NGB) overexpressed inEscherichia coli(E. coli) bacteria. The feasibility of the setup regarding sensitivity and photo-induced effects and the results regarding the oxygen uptake and release will be discussed and compared for each system. The summary of the results show that the method is promising and the setup will be developed further to monitor the dependence of the neuronal action potential on nerve globins.
Highlights
Ligand-binding globins are found throughout all kingdoms of nature
Ramser et al / The combination of resonance Raman spectroscopy, optical tweezers and microfluidic systems 289 the aspiration to investigate the protein in functional cells under physiological-like conditions
If the spectrometer was equipped with an upright microscope, a double microscope configuration was chosen to enable the integration of the optical trap together with the microfluidic system [21], meaning that the optical path for the optical tweezers and the Raman spectrometer were separated
Summary
Ligand-binding globins are found throughout all kingdoms of nature. Various functions are identified, such as oxygen delivery and storage and enzymatic activities. The authors suggest that TrxR and NGB belong to the defence system of the nerve cell against hypoxia and the oxidative damage occurring by reoxygenation This suggestion demands that the neuroglobins are able to form disulfide bridges, which so far only has been shown in vitro [11]. Even though first successful in vivo experiments performed on mice have been reported [6], further tools need to be developed to study functional heme-containing cells under varying physiological conditions in order to elucidate the mechanisms of cell protection against oxidative stress. K. Ramser et al / The combination of resonance Raman spectroscopy, optical tweezers and microfluidic systems 289 the aspiration to investigate the protein in functional cells under physiological-like conditions. In this article we will summarize and compare the results of our studies and report about the further development of the technique that will enable to study the dependence of the action potential of neuronal cells and nerve cords upon heme proteins
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 call
