TY - JOUR
T1 - Monitoring neurochemical release from astrocytes using in vitro microdialysis coupled with high-speed capillary electrophoresis
AU - Hogerton, Amy L.
AU - Bowser, Michael T.
PY - 2013/10/1
Y1 - 2013/10/1
N2 - 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.
AB - 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.
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U2 - 10.1021/ac401631k
DO - 10.1021/ac401631k
M3 - Article
C2 - 23984889
AN - SCOPUS:84884995015
SN - 0003-2700
VL - 85
SP - 9070
EP - 9077
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 19
ER -