title: Monitoring neurochemical release from astrocytes using in vitro microdialysis coupled with high-speed capillary electrophoresis.
abstract: We have developed a novel in vitro approach for monitoring fast neurochemical dynamics in model cell systems using microdialysis sampling coupled with high-speed capillary electrophoresis (CE). Cells from an immortalized astrocyte line (C8-D1A) were cultured in direct contact with the porous membrane of a microdialysis probe. Confocal microscopy was used to confirm cell viability and confluency over the microdialysis sampling region. Small molecules released from the astrocytes were efficiently sampled by the probe due to the direct contact with the membrane. Microdialysis sampling was coupled with online, high-speed CE allowing changes in the dialysate concentration of small molecule amine neurochemicals to be monitored with 20 s temporal resolution. Basal release of a number of important analytes was detected including glycine, taurine, D-serine, and glutamate. The ability of the in vitro microdialysis-CE instrument to monitor dynamic changes in analyte concentration was assessed by transferring a probe cultured with astrocytes from a solution containing artificial cerebrospinal fluid (aCSF) to a high K(+) solution (100 mM K(+)-aCSF). Upon stimulation, the observed concentration of a number of key neurochemicals increased dramatically including glycine (700%), taurine (185%), and serine (215%). Amino acids such as phenylalanine and valine, which are not known to respond to cellular swelling mechanisms, were unaffected by the K(+) stimulation.
authors.affiliation: Department of Chemistry, University of Minnesota , 207 Pleasant St. SE, Minneapolis, Minnesota 55455, United States.
authors.forename: Amy L
authors.initials: AL
authors.lastname: Hogerton
authors.forename: Michael T
authors.initials: MT
authors.lastname: Bowser
mesh_terms: D000596:Amino Acids / Q000032:analysis / Q000378:metabolism; D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism*; D002470:Cell Survival; D002478:Cells, Cultured; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D005998:Glycine / Q000032:analysis / Q000378:metabolism; D051379:Mice; D017551:Microdialysis / Q000295:instrumentation / Q000379:methods*; D018377:Neurotransmitter Agents / Q000032:analysis* / Q000378:metabolism*; D012694:Serine / Q000032:analysis / Q000378:metabolism; D013654:Taurine / Q000032:analysis / Q000378:metabolism
chemical_list: D000596:Amino Acids; D018377:Neurotransmitter Agents; D013654:Taurine; D012694:Serine; D005998:Glycine