Real-time evaluation of nitric oxide (NO) levels in cortical and hippocampal areas with a nanopore-based electrochemical NO sensor

Nitric oxide (NO) is an important biomolecule for regulating various brain functions, such as the control of neurovascular tone. NO, however, cannot be stored inside cells where NO is produced and immediately diffuses through the cellular membrane and decays rapidly, which makes the detection of NO extremely hard in an in vivo setting. We constructed an amperometric NO nanosensor and utilized it to directly measure NO release in the living brain. The NO nanosensor uses nanopores (pores with an opening radii <500 nm) in which NO is oxidized at the porous platinum surface. The nanopore-based sensor was inserted vertically into the brains of anesthetized mice up to the end of the hippocampal CA 3 region, or to a depth of about 3 mm. The sensor was slowly advanced in the brain in 0.5 μm increments and in 0.05 s temporal steps. Different levels of NO release were monitored by the nanopore NO sensor during the course of the penetration. The hippocampal CA3 region had the highest level of NO release, which was followed by CA2 and CA1 of the hippocampus and the cortex. The levels of NO release were not uniformly distributed within the cortical and hippocampal areas of living brain. In sum, the nanopore-based NO sensor was able to grossly measure NO contents within living brain in real time and with high sensitivity. This study may provide good insights about the relationship between the distributions of NOS-immunoreactive neurons and the directly measured levels of NO release in brain.