In vivo spectroscopy: a novel approach for simultaneously estimating nitric oxide and hemodynamic parameters in the rat brain

Abstract

Nitric oxide (NO) is a versatile molecule involved in a wide range of biological processes. Under physiological conditions, NO reacts with oxyhemoglobin (OxyHb) to form methemoglobin (MetHb) at a very high rate. Microdialysis studies have used hemoglobin solutions as a trapping method to quantify NO in vivo. The methodology described here uses the microcapillary network with endogenous OxyHb instead of microdialysis probe with exogenous OxyHb for monitoring MetHb as an indirect index of NO levels by in vivo spectroscopy using optical fibers. This new method has been validated in rat cerebral cortex by the infusion of NO or well-known drug-induced changes in NO concentration (NMDA agonists and a NO-synthase inhibitor) and by comparing results with simultaneous voltammetric recordings. Results indicate that this spectroscopy technique is able to record large increases in MetHb levels and to detect reductions of its basal levels. In addition, data show that similar changes and kinetics can be observed with both techniques. Thus, intravascular MetHb can be used as an indirect index of NO levels. It is proposed that in vivo spectroscopy may be a useful tool to gain insight into the roles of NO in hemodynamic parameters and in other physiological processes such as the regulation of the mitochondrial respiratory chain.