 |
 Previous Article | Next Article 
Journal of Neuroscience, Vol 9, 38-48, Copyright © 1989 by Society for Neuroscience
Aplysia synaptosomes. I. Preparation and biochemical and morphological characterization of subcellular membrane fractions
GJ Chin, E Shapiro, SS Vogel and JH Schwartz
Howard Hughes Medical Institute, Center for Neurobiology and Behavior, Columbia University, New York, New York 10032.
We prepared and characterized subcellular membrane fractions from the CNS
of Aplysia californica that are enriched in isolated nerve terminals
(synaptosomes). Ganglia were homogenized in 1.1 M sucrose and fractionated
on a 2-step sucrose gradient, yielding 50 micrograms protein/animal in the
synaptosomal fraction (P3), which was enriched 3- fold in plasma membrane
as compared with the initial homogenate. Quantitative morphometry of
electron micrographs revealed that P3 contained 25% intact synaptosomes, a
5-fold enrichment over the homogenate. Although fractionation on a 5-step
sucrose gradient reduced the yield of protein in the synaptosomal fraction
to 40 micrograms/animal, this fraction (the 0.35 M/0.75 M interface) was
more enriched in plasma membrane than P3 and was less contaminated by
lysosomes and free mitochondria. By electron microscopy, the 0.35 M/0.75 M
interface contained up to 50% synaptosomes. Synaptosomal fractions
contained cAMP-, Ca2+/calmodulin-, and Ca2+/phospholipid- dependent protein
kinase activities and were enriched in a Mr 40,000 pertussis toxin
substrate, Gi/o. In the accompanying paper, we show that these synaptosomes
retain the ability to release transmitters.
This article has been cited by other articles:
|
|
|
|
 
D. Fioravante, R.-Y. Liu, and J. H. Byrne
The Ubiquitin-Proteasome System Is Necessary for Long-Term Synaptic Depression in Aplysia
J. Neurosci.,
October 8, 2008;
28(41):
10245 - 10256.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Y. Yao, M. T. Kelly, S. Sajikumar, P. Serrano, D. Tian, P. J. Bergold, J. U. Frey, and T. C. Sacktor
PKM{zeta} Maintains Late Long-Term Potentiation by N-Ethylmaleimide-Sensitive Factor/GluR2-Dependent Trafficking of Postsynaptic AMPA Receptors
J. Neurosci.,
July 30, 2008;
28(31):
7820 - 7827.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Carroll, J. Dyer, and W. S. Sossin
Serotonin Increases Phosphorylation of Synaptic 4EBP through TOR, but Eukaryotic Initiation Factor 4E Levels Do Not Limit Somatic Cap-Dependent Translation in Aplysia Neurons
Mol. Cell. Biol.,
November 15, 2006;
26(22):
8586 - 8598.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
O. Khabour, J. Levenson, L. C. Lyons, L. S. Kategaya, J. Chin, J. H. Byrne, and A. Eskin
Coregulation of Glutamate Uptake and Long-Term Sensitization in Aplysia
J. Neurosci.,
October 6, 2004;
24(40):
8829 - 8837.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Liu, J.-Y. Hu, S. Schacher, and J. H. Schwartz
The Two Regulatory Subunits of Aplysia cAMP-Dependent Protein Kinase Mediate Distinct Functions in Producing Synaptic Plasticity
J. Neurosci.,
March 10, 2004;
24(10):
2465 - 2474.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Khan, A. M. Pepio, and W. S. Sossin
Serotonin Activates S6 Kinase in a Rapamycin-Sensitive Manner in Aplysia Synaptosomes
J. Neurosci.,
January 15, 2001;
21(2):
382 - 391.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Levenson, S. Endo, L. S. Kategaya, R. I. Fernandez, D. G. Brabham, J. Chin, J. H. Byrne, and A. Eskin
Long-term regulation of neuronal high-affinity glutamate and glutamine uptake in Aplysia
PNAS,
October 23, 2000;
(2000)
220256497.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
P. Skehel, K. Martin, E. Kandel, and D Bartsch
A VAMP-binding protein from Aplysia required for neurotransmitter release
Science,
September 15, 1995;
269(5230):
1580 - 1583.
[Abstract]
[PDF]
|
|
|
|
|
|
|
J. Levenson, S. Endo, L. S. Kategaya, R. I. Fernandez, D. G. Brabham, J. Chin, J. H. Byrne, and A. Eskin
Long-term regulation of neuronal high-affinity glutamate and glutamine uptake in Aplysia
PNAS,
November 7, 2000;
97(23):
12858 - 12863.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
