Analysis of D-Aspartate as a Signaling Molecule in the Aplysia Californica Central Nervous System Using Capillary Electrophoresis and Radioisotopic Labeling
Scanlan, Cory Randolph
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https://hdl.handle.net/2142/84251
Description
Title
Analysis of D-Aspartate as a Signaling Molecule in the Aplysia Californica Central Nervous System Using Capillary Electrophoresis and Radioisotopic Labeling
Author(s)
Scanlan, Cory Randolph
Issue Date
2006
Doctoral Committee Chair(s)
Sweedler, Jonathan V.
Department of Study
Chemistry
Discipline
Chemistry
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Chemistry, Analytical
Language
eng
Abstract
A second cell-cell signaling molecule investigated is D-aspartate (D-Asp). D-Asp is present in the nervous systems of both vertebrates and invertebrates, and its biosynthesis has been observed in mammalian cells. We suspect that D-Asp acts as a classical neurotransmitter in the central nervous system. The mollusk Aplysia californica has shown high levels of D-Asp in the neural ganglia; and this, along with its relatively simple invertebrate system, makes Aplysia a viable model system for studying the function and neurochemistry of D-Asp. Using Aplysia californica , we have investigated the localization of D-Asp as well as its synthesis from L-Asp. We developed a technique utilizing off-line CE coupled with radionuclide detection, which enables high sensitivity characterization of L- to D-Asp conversion. Furthermore, these capillary electrophoretic techniques, along with liquid scintillation counting and MALDI-MS analysis, have allowed us to address other criteria required to determine whether D-Asp acts as a classical neurotransmitter, including release from cells, sodium dependence of uptake, response to D-Asp by tissues, and the D-Asp stimulated release of cardiomodulatory peptides from the R3-14 cells. These techniques developed provide robust methods for the analysis of neurotransmitters and elucidation of their chiral moieties in complex mass-limited biological samples.
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