title: A novel method for simultaneous glutamate and extracellular activity measurement in brain slices with high temporal resolution.
abstract: Measurement of neurotransmitters during normal or altered function in cerebral slices could be an important tool to better understand the relationship between biochemical changes and electrophysiological activity. Some attempts of this analysis have been made; however, the current techniques do not have the appropriate time resolution to establish this relationship. The use of electrochemical biosensors has allowed for good time resolution, but problems related to the reduction of signal noise and biofouling of the electrode surface could be an important issue. In this work, we propose a new alternative to simultaneously measure glutamate and electrical activity with a high temporal resolution in brain slices. This approach is based on the use of enzymatic reactors that generate a fluorescent derivative from glutamate that can be measured at high temporal resolution. The results presented here show a reliable measurement of this neurotransmitter in brain slices obtained from intact animals under the effect of a glutamate transporter blocker DL-threo-beta-benzyloxyaspartate as well as the potassium channel blocker 4-aminopyridine. Differences in the levels of glutamate and high frequency and amplitude discharges as an effect of drug administration were found in brain slices obtained from epileptic rats (p<0.05). In conclusion, this method could be used to measure neurotransmitter concentration online at a near physiological temporal resolution, which can then be correlated to the electrical activity that is simultaneously recorded.
authors.affiliation: Laboratory of Neurophysiology and Neurochemistry, Department of Cellular and Molecular Biology, CUCBA, University of Guadalajara, Jalisco, Mexico.
authors.forename: C L
authors.initials: CL
authors.lastname: López-Valenzuela
authors.affiliation: Laboratory of Neurophysiology and Neurochemistry, Department of Cellular and Molecular Biology, CUCBA, University of Guadalajara, Jalisco, Mexico. Electronic address: amorales@cucba.udg.mx.
authors.forename: A
authors.initials: A
authors.lastname: Morales-Villagrán
authors.affiliation: Laboratory of Neurophysiology and Neurochemistry, Department of Cellular and Molecular Biology, CUCBA, University of Guadalajara, Jalisco, Mexico.
authors.forename: L
authors.initials: L
authors.lastname: Medina-Ceja
mesh_terms: D015761:4-Aminopyridine / Q000494:pharmacology; D027322:Amino Acid Transport System X-AG / Q000037:antagonists & inhibitors; D000818:Animals; D001224:Aspartic Acid / Q000494:pharmacology; D001921:Brain / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*; D004594:Electrophysiology / Q000295:instrumentation / Q000379:methods*; D005110:Extracellular Space / Q000187:drug effects / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry; D018698:Glutamic Acid / Q000737:chemistry / Q000378:metabolism*; D006624:Hippocampus / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D008297:Male; D051381:Rats; D017208:Rats, Wistar
chemical_list: D027322:Amino Acid Transport System X-AG; D005456:Fluorescent Dyes; C120673:benzyloxyaspartate; D001224:Aspartic Acid; D018698:Glutamic Acid; D015761:4-Aminopyridine
keywords: Brain slices; Extracellular recording; Glutamate; High temporal resolution; Online measurement