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QUANTITATIVE DETERMINATION OF ECTOPEPTIDASE ACTIVITY IN THE RAT HIPPOCAMPUS USING ELECTROOSMOTIC PUSH-PULL PERFUSION COUPLED TO CAPILLARY LIQUID CHROMATOGRAPHY: A COMPUTATIONAL AND EXPERIMENTAL APPROACH

Ou, Yangguang (2018) QUANTITATIVE DETERMINATION OF ECTOPEPTIDASE ACTIVITY IN THE RAT HIPPOCAMPUS USING ELECTROOSMOTIC PUSH-PULL PERFUSION COUPLED TO CAPILLARY LIQUID CHROMATOGRAPHY: A COMPUTATIONAL AND EXPERIMENTAL APPROACH. Doctoral Dissertation, University of Pittsburgh. (Unpublished)

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Abstract

It has been known for over a century that the CA1 subregion of the hippocampus is more vulnerable than the CA3 to ischemic damage. While many studies have been conducted the exact mechanism is still unknown. Ectopeptidases are membrane-bound enzymes whose catalytic domains face the extracellular space. Traditionally, their role was believed to only involve clearance of active peptides but recent studies have shown that they play an important role in regulating peptide activity. Here we report the quantitative measurement of ectopeptidase activity using electroosmotic push-pull perfusion coupled to offline capillary liquid chromatography. This method revealed a three-fold higher aminopeptidase activity hydrolyzing the neuroprotective peptide Leu-enkephalin in the CA1 region of the rat hippocampus. Inhibition of the higher aminopeptidase activity in the CA1 selectively protects this region from ischemic damage due to oxygen-glucose deprivation. This is the first report of spatially-resolved quantitative measurements of enzyme activity in intact tissue using native substrates. The studies in this thesis combine both computational and experimental approaches to tackle a longstanding question in neuroscience.


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Details

Item Type: University of Pittsburgh ETD
Status: Unpublished
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Ou, Yangguangyao5@pitt.eduyao50000-0002-6902-3978
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Committee ChairWeber, Stephensweber@pitt.edusweber
Committee MemberMichael, Adrianamichael@pitt.eduamichael
Committee MemberChilders, W.wschild@pitt.eduwschild
Committee MemberBarrionuevo, Germangerman@pitt.edugerman
Date: 31 January 2018
Date Type: Publication
Defense Date: 1 December 2017
Approval Date: 31 January 2018
Submission Date: 7 November 2017
Access Restriction: No restriction; Release the ETD for access worldwide immediately.
Number of Pages: 142
Institution: University of Pittsburgh
Schools and Programs: Dietrich School of Arts and Sciences > Chemistry
Degree: PhD - Doctor of Philosophy
Thesis Type: Doctoral Dissertation
Refereed: Yes
Uncontrolled Keywords: ectopeptidase, enzyme activity, ischemia, push-pull perfusion, organotypic slice culture, finite element
Date Deposited: 31 Jan 2018 18:38
Last Modified: 31 Jan 2018 18:38
URI: http://d-scholarship.pitt.edu/id/eprint/33322

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