• Login
    View Item 
    •   UMB Digital Archive
    • School, Graduate
    • Theses and Dissertations All Schools
    • View Item
    •   UMB Digital Archive
    • School, Graduate
    • Theses and Dissertations All Schools
    • View Item
    JavaScript is disabled for your browser. Some features of this site may not work without it.

    Browse

    All of UMB Digital ArchiveCommunitiesPublication DateAuthorsTitlesSubjectsThis CollectionPublication DateAuthorsTitlesSubjects

    My Account

    LoginRegister

    Statistics

    Display statistics

    The Role of Testisin in Endothelial Cells and Angiogenesis

    • CSV
    • RefMan
    • EndNote
    • BibTex
    • RefWorks
    Thumbnail
    Name:
    Peroutka_umaryland_0373D_11261.pdf
    Size:
    9.100Mb
    Format:
    PDF
    Download
    Author
    Peroutka, Raymond cc
    0000-0002-8639-5773
    Advisor
    Antalis, Toni M.
    Date
    2021
    Type
    dissertation
    
    Metadata
    Show full item record
    Abstract
    Testisin (PRSS21) is a membrane anchored serine protease, which is tethered to the cell surface via a glycosylphosphatidylinositol (GPI)-anchor. Testisin expression has been documented in eosinophiles, ovarian cancers, endothelial cells, and spermatozoa where its expression is highest. Although two substrates of testisin have so far been identified, protease activated receptor 2 (PAR2) and protein inhibitor C (PCI), little is known of the biological, physiological, and pathophysiologic characteristics of testisin. Thus far, there are no published data identifying an activator or inhibitor of testisin. To better characterize the biochemistry of testisin we produced the zymogen as inclusion bodies in E. coli and refolded using the insoluble cellular fraction. To better characterize the cellular functions of testisin, hybridomas producing anti-testisin monoclonal antibodies were acquired, antibodies purified, and then characterized. In an investigation of testisin’s function in endothelial cells we identified testisin as a novel regulator of physiological hormone-induced angiogenesis and microvascular endothelial permeability. Using a murine model of rapid physiological angiogenesis during corpus luteal development in the ovary, we found that mice genetically deficient in testisin (Prss21-/-) show a substantially increased incidence of hemorrhages which are significantly more severe than in littermate control Prss21+/+ mice. This phenotype was associated with increased vascular leakiness, demonstrated by a greater accumulation of extravasated Evans blue dye in Prss21-/- ovaries. Live cell imaging of in vitro cultured microvascular endothelial cells depleted of testisin by siRNA knockdown revealed that loss of testisin markedly impaired reorganization and tubule-like formation on Matrigel. Moreover, testisin siRNA knockdown increased the paracellular permeability to FITC-albumin across endothelial cell monolayers, which was associated with decreased expression of the adherens junction protein VE-cadherin and increased levels of phospho-(Tyr658)-VE-cadherin, without affecting the levels of the tight junction proteins occludin, claudin-5, or ZO-1. Decreased expression of VE-cadherin in the neovasculature of Prss21-/- ovaries was also observed without marked differences in endothelial cell content, vascular claudin-5 expression or pericyte recruitment. Together, these data identify testisin as a novel regulator of VE-cadherin adhesions during angiogenesis and indicate a potential new target for regulating neovascular integrity and associated pathologies.
    Description
    Molecular Medicine
    University of Maryland, Baltimore
    Ph.D.
    Keyword
    testisin
    Blood-vessels--Growth
    Endothelial Cells
    Identifier to cite or link to this item
    http://hdl.handle.net/10713/16528
    Collections
    Theses and Dissertations School of Medicine
    Theses and Dissertations All Schools

    entitlement

     
    DSpace software (copyright © 2002 - 2023)  DuraSpace
    Quick Guide | Policies | Contact Us | UMB Health Sciences & Human Services Library
    Open Repository is a service operated by 
    Atmire NV
     

    Export search results

    The export option will allow you to export the current search results of the entered query to a file. Different formats are available for download. To export the items, click on the button corresponding with the preferred download format.

    By default, clicking on the export buttons will result in a download of the allowed maximum amount of items.

    To select a subset of the search results, click "Selective Export" button and make a selection of the items you want to export. The amount of items that can be exported at once is similarly restricted as the full export.

    After making a selection, click one of the export format buttons. The amount of items that will be exported is indicated in the bubble next to export format.