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Journal of Neuroscience Methods 154 (2006) 245255 Filtration disrupts synaptosomes during radiochemical analysis
 

Summary: Journal of Neuroscience Methods 154 (2006) 245255
Filtration disrupts synaptosomes during radiochemical analysis
of serotonin uptake: Comparison with chronoamperometry
in SERT knockout mice
Xiomara A. Pereza, Laura E. Biancoa, Anne M. Andrewsb,c,
a Department of Chemistry, The Pennsylvania State University, University Park, PA 16802, United States
b Huck Institutes of the Life Sciences, 201 Life Science Building, The Pennsylvania State University, University Park, PA 16802, United States
c Department of Veterinary and Biomedical Sciences, The Pennsylvania State University, University Park, PA 16802, United States
Received 31 January 2005; received in revised form 4 December 2005; accepted 23 December 2005
Abstract
Radiochemical methods have failed to reveal decreases in synaptosomal serotonin uptake in mice lacking one functional copy of the serotonin
transporter (SERT) gene. By contrast, uptake rates determined by chronoamperometry in synaptosomes from SERT+/- mice show gene-related
reductions. We revisited [3
H]5-HT uptake in SERT knockout mice to determine the effects of inclusion of O2 in the incubation buffer on the kinetic
parameters obtained by this method. In oxygenated synaptosomes prepared from frontal cortex and striatum, modest 25 and 35% reductions in
radiolabeled 5-HT uptake were detected in SERT+/- versus SERT+/+ mice. However, even in the presence of O2, no differences in [3
H]5-HT
uptake were detected between SERT+/- and SERT+/+ mice in brain stem in contrast to 60% reductions determined by chronoamperometry.
Moreover, while inclusion of O2 modestly increased the rates of [3
H]5-HT uptake, rates determined by chronoamperometry in the presence of O2

  

Source: Andrews, Anne M. - Huck Institutes of the Life Sciences, Pennsylvania State University

 

Collections: Biology and Medicine