Addressing the Neurochemical Problem: Sensitive and Selective Measurements of Neurotransmitters, Neuropeptides, and Synaptic Vesicles

Persistent Link:
http://hdl.handle.net/10150/556979
Title:
Addressing the Neurochemical Problem: Sensitive and Selective Measurements of Neurotransmitters, Neuropeptides, and Synaptic Vesicles
Author:
Laude, Nicholas D.
Issue Date:
2015
Publisher:
The University of Arizona.
Rights:
Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
Abstract:
The neurochemical problem (1) and the directive of the neuroanalytical chemist (2) can be stated as follows: (1) The chemical space of the nervous system is populated by hundreds of neuroactive species linked through extensive biological circuits which are dynamically changing in time and space in response to myriad inputs. (2) Neurochemical analysis techniques should therefore have the appropriate temporal, spatial, and chemical resolution to study these systems, while perturbing them so minimally as to allow unfettered in vivo measurements. New tools and concepts for analytical measurements of neurotransmitters, neuropeptides, and synaptic vesicles are developed and presented in this dissertation on analytical measurements for addressing the neurochemical problem. The introduction gives a broad overview of chemical neuroscience and introduces quantitative visualization of the multidimensional resolution paradigm for analytical chemists seeking to design effective experiments. Chapters two through four detail advancements in data processing and instrument design which decrease detection limits and allow for improved spatial, temporal, chemical resolution in capillary electrophoresis measurements of neurotransmitters, metabolites, and synaptic vesicles. Chapter five discusses the development of fast-scan controlled-adsorption voltammetry which has dramatically increases the spatial and temporal resolution of basal dopamine measurement in vivo. Chapter six introduces online-preservation microdialysis as a way to overcome enzymatic degradation of endogenous opioid neuropeptides during in vivo sample collection. Because of this discovery of the secretory behavior these neuropeptides is reported in the anterior cingulate cortex (ACC), a region of the brain deeply associated with pain signaling. The advancement of peptide drugs particularly glycosylated neuropeptide analogs through new methods of mass spectrometry analysis for rapid feedback in drug development are presented in chapter 7. Chapter eight concludes this work with future directions pointing towards single-cell electrochemical and mass spectrometry measurements, shotgun-microdialysis for high-throughput screening of neurotherapeutics, preliminary data on the effect of chronic pain of endogenous opioids in the ACC, and the beginnings of in vivo neuroproteomics analysis in rodent pain models.
Type:
text; Electronic Dissertation
Keywords:
Enkephalins; Mass Spectrometry; Microdialysis; Neurochemical; Peptides; Chemistry; Capillary Electrophoresis
Degree Name:
Ph.D.
Degree Level:
doctoral
Degree Program:
Graduate College; Chemistry
Degree Grantor:
University of Arizona
Advisor:
Heien, Michael L.

Full metadata record

DC FieldValue Language
dc.language.isoen_USen
dc.titleAddressing the Neurochemical Problem: Sensitive and Selective Measurements of Neurotransmitters, Neuropeptides, and Synaptic Vesiclesen_US
dc.creatorLaude, Nicholas D.en
dc.contributor.authorLaude, Nicholas D.en
dc.date.issued2015en
dc.publisherThe University of Arizona.en
dc.rightsCopyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.en
dc.description.abstractThe neurochemical problem (1) and the directive of the neuroanalytical chemist (2) can be stated as follows: (1) The chemical space of the nervous system is populated by hundreds of neuroactive species linked through extensive biological circuits which are dynamically changing in time and space in response to myriad inputs. (2) Neurochemical analysis techniques should therefore have the appropriate temporal, spatial, and chemical resolution to study these systems, while perturbing them so minimally as to allow unfettered in vivo measurements. New tools and concepts for analytical measurements of neurotransmitters, neuropeptides, and synaptic vesicles are developed and presented in this dissertation on analytical measurements for addressing the neurochemical problem. The introduction gives a broad overview of chemical neuroscience and introduces quantitative visualization of the multidimensional resolution paradigm for analytical chemists seeking to design effective experiments. Chapters two through four detail advancements in data processing and instrument design which decrease detection limits and allow for improved spatial, temporal, chemical resolution in capillary electrophoresis measurements of neurotransmitters, metabolites, and synaptic vesicles. Chapter five discusses the development of fast-scan controlled-adsorption voltammetry which has dramatically increases the spatial and temporal resolution of basal dopamine measurement in vivo. Chapter six introduces online-preservation microdialysis as a way to overcome enzymatic degradation of endogenous opioid neuropeptides during in vivo sample collection. Because of this discovery of the secretory behavior these neuropeptides is reported in the anterior cingulate cortex (ACC), a region of the brain deeply associated with pain signaling. The advancement of peptide drugs particularly glycosylated neuropeptide analogs through new methods of mass spectrometry analysis for rapid feedback in drug development are presented in chapter 7. Chapter eight concludes this work with future directions pointing towards single-cell electrochemical and mass spectrometry measurements, shotgun-microdialysis for high-throughput screening of neurotherapeutics, preliminary data on the effect of chronic pain of endogenous opioids in the ACC, and the beginnings of in vivo neuroproteomics analysis in rodent pain models.en
dc.typetexten
dc.typeElectronic Dissertationen
dc.subjectEnkephalinsen
dc.subjectMass Spectrometryen
dc.subjectMicrodialysisen
dc.subjectNeurochemicalen
dc.subjectPeptidesen
dc.subjectChemistryen
dc.subjectCapillary Electrophoresisen
thesis.degree.namePh.D.en
thesis.degree.leveldoctoralen
thesis.degree.disciplineGraduate Collegeen
thesis.degree.disciplineChemistryen
thesis.degree.grantorUniversity of Arizonaen
dc.contributor.advisorHeien, Michael L.en
dc.contributor.committeememberHeien, Michael L.en
dc.contributor.committeememberArmstrong, Neal R.en
dc.contributor.committeememberAspinwall, Craig A.en
dc.contributor.committeememberBrown, Michael F.en
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