Feasibility of continuous glutamate monitoring in perfused retinal tissue with a potentiometric biosensing probe.
A potentiometric biosensing probe for glutamate has been evaluated as a possible tool to measure the release of glutamate from the isolated retina of Bufo marinus. This probe is based on carbon dioxide detection, following enzymatic conversion of glutamate to gamma-aminobutyric acid (GABA) via glutamic acid decarboxylase (GAD). Probe response characteristics of dynamic range, limit of detection, pH dependency, and selectivity are described. Probe modifications were required for sensor operation in an upside down configuration which was demanded by the need to mount and perfuse the retinal tissue directly at the sensor tip. Overall, these results indicate that this particular potentiometric biosensor is not well suited for direct glutamate measurements in retinal tissue because of pH incompatibility between the sensor and the tissue, and because of high background carbon dioxide levels released from the retina at the pH optimum of the probe. Despite this drawback, the sensor could be utilized to provide a continuous "downstream" monitor of glutamate levels during the course of an experiment, after pH buffering of tissue perfusate. Alternative approaches to probes more compatible with direct tissue measurements are discussed.
D000818:Animals; D002024:Bufo marinus; D005971:Glutamates / Q000378:metabolism*; D018698:Glutamic Acid; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007202:Indicators and Reagents; D010477:Perfusion / Q000295:instrumentation / Q000379:methods; D011199:Potentiometry; D012160:Retina / Q000378:metabolism*
D005971:Glutamates; D007202:Indicators and Reagents; D018698:Glutamic Acid