TY - JOUR
T1 - Kinetic and quantitative measurements of catecholamine transport in chromaffin ghosts using a glassy carbon electrode
AU - Hayflick, Susan
AU - Johnson, Robert G.
AU - Carty, Sally E.
AU - Scarpa, Antonio
N1 - Funding Information:
Many thanks to Dr. Curt Freed for invaluable assistance with the initial experiments and for his continuous suggestions and advice, to Dr. Jack Leigh for a criticat reading of the manuscript, and to Dan Brannen for preparation of the manuscript. This work was supported by Grants HL-18708 and HL-240 10 from the National Institutes of Health.
PY - 1982/10
Y1 - 1982/10
N2 - A method for the on-line kinetic measurement of net catecholamine uptake and release in ghosts derived from bovine chromaffin granules is described. Changes in free catecholamine concentration in 1 to 2 ml of media containing chromaffin ghosts were continuously measured through the amperometric detection of their oxidation products through a glassy carbon electrode set at 0.5-V potential vs a reference electrode. Parallel measurements of catecholamine uptake and release in the ghosts under various metabolic conditions show a good quantitative agreement between the values obtained with the electrode and those obtained through high-performance liquid chromatography after separation of the ghosts from the medium. Initial velocities of ATP-dependent uptake of epinephrine, norepinephrine, dopamine, and isoproternol by the ghosts are shown. This method permits, for the first time, quantitation of unidirectional movement of catecholamines in the presence of minute quantities of biological samples. The advantages, limitations, and suitability of this method to measure catecholamine transport in other systems are discussed.
AB - A method for the on-line kinetic measurement of net catecholamine uptake and release in ghosts derived from bovine chromaffin granules is described. Changes in free catecholamine concentration in 1 to 2 ml of media containing chromaffin ghosts were continuously measured through the amperometric detection of their oxidation products through a glassy carbon electrode set at 0.5-V potential vs a reference electrode. Parallel measurements of catecholamine uptake and release in the ghosts under various metabolic conditions show a good quantitative agreement between the values obtained with the electrode and those obtained through high-performance liquid chromatography after separation of the ghosts from the medium. Initial velocities of ATP-dependent uptake of epinephrine, norepinephrine, dopamine, and isoproternol by the ghosts are shown. This method permits, for the first time, quantitation of unidirectional movement of catecholamines in the presence of minute quantities of biological samples. The advantages, limitations, and suitability of this method to measure catecholamine transport in other systems are discussed.
UR - http://www.scopus.com/inward/record.url?scp=0020460548&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0020460548&partnerID=8YFLogxK
U2 - 10.1016/0003-2697(82)90108-7
DO - 10.1016/0003-2697(82)90108-7
M3 - Article
C2 - 7181117
AN - SCOPUS:0020460548
SN - 0003-2697
VL - 126
SP - 58
EP - 66
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 1
ER -