PMID: 11107168Dec 7, 2000

Isolation and characterization of substance P-containing dense core vesicles from rabbit optic nerve and termini

Journal of Neuroscience Research
E A BergR E Fine

Abstract

In neurons, neuropeptides and other synaptic components are transported down the axon to the synapse in vesicles using molecular motors of the kinesin family. In the synapse, these neuropeptides are found in dense core vesicles (DCVs), and, following calcium-mediated exocytosis, they interact with receptors on the target cell. We have developed a rapid, large-scale technique for purifying peptide-containing DCVs from specific nuclei in the central nervous system. By using differential velocity gradient and equilibrium gradient centrifugation, neuropeptide-containing DCVs can be separated by size and density from optic nerve (ON) and its termini, the lateral geniculate nuclei and the superior colliculi. Isolated DCVs contain neuropeptides (substance P and brain-derived neurotrophic factor), synaptic vesicle (SV) membrane proteins (SV2, synaptotagmins, synaptophysin, Rab3 and synaptobrevin), SV-associated proteins (alpha-synuclein), secretory markers for DCVs previously isolated (secretogranin II), and beta-amyloid precursor protein. By using electron microscopic techniques, DCV were also visualized and shown to be immunoreactive for neuropeptides, neurotrophins, and SV membrane proteins. Because of the interesting group of physi...Continue Reading

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Related Concepts

TAC1
SNCA gene
Calcium [EPC]
Optic Lobe, Human
Entire Optic Nerve
Calcium
Neurons
Amyloid Beta Precursor Protein Measurement
Recombinant Brain-Derived Growth Factors
Synaptic Vesicles

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