Fiber-optic ammonia sensor for measuring synaptic glutamate and extracellular ammonia

A fiber-optic ammonia gas sensor designed for neurochemical applications is presented. Parameters evaluated in terms of effect on the steady-state and dynamic response of this sensor include the indicator dye, concentrations of indicator and total ammonia nitrogen in the internal solution, volume of the internal solution, structure of the gas-permeable membrane, and temperature. The final ammonia sensor responds over the concentration range from 7 to 3000 nM with a limit of detection of 7 nM and response times ranging from 2 to 5 min. Glutamate oxidase is immobilized at the tip of this ammonia sensor to provide a glutamate biosensor with a detection limit of 0.1 microM when operated at pH 7.8. In addition, this ammonia sensor is used to measure extracellular ammonia levels in perfused retinal and eye-cup tissue preparations. These measurements indicate a calcium-dependent, potassium-evoked release of ammonia during these depolarization conditions.