عنوان به زبان ديگر :
Cerebellar Giant Synaptosomes: a Model to Study Basal and Stimulated Release of [3H] gamma-Aminobutyric Acid
پديد آورندگان :
Hosseini S. M. نويسنده , ASGARI A. نويسنده , Mehrani H. A. نويسنده , KHOSHBATEN A. نويسنده
چكيده لاتين :
Background: Neurotransmitter release is an essential link in
cell communication of the nervous system. Many investigations
have focused on gamma amino butyric acid (GABA)ergic
neurotransmission, because it has been implicated in the
pathophysiology of several central nervous system disorders.
To bypass complications related to homo- and heterosynaptic
modulation and to avoid indirect interpretations of data, we
herein describe a simple approach for direct measurement of
GABA release.
Material and Methods: Giant synaptosomes originated from
nerve terminals of rat cerebellum mossy fibers were prepared
for the study. Electron micrographs as well as lactate dehydrogenase
assay are used for morphological and biochemical verifications.
Giant synaptosomes were preloaded with labeled
[3H]GABA. Spontaneous and stimulated release of eH]GABA
was measured using a superfusion apparatus. Stimulation was
evoked by increasing exteracellular concentration of K+ ions.
Results: Spontaneous [3H]GABA release had a constant and
nearly linear kinetics. eH]GABA outflow evoked by depolarizing
solution containing 15 mM of K+ showed 2-3 fold increases
over the basal release. The same effect was also reproducible
after several minutes.
Conclusion: The present findings indicate that this preparation
could be used as a suitable and versatile in vitro model to
study GABA release from axon terminals under basal and
evoked conditions.