• Find People
  • Campus Map
  • PiratePort
  • A-Z
    • About
    • Submit
    • Browse
    • Login
    View Item 
    •   ScholarShip Home
    • ECU Main Campus
    • Thomas Harriot College of Arts and Sciences
    • Biology
    • View Item
    •   ScholarShip Home
    • ECU Main Campus
    • Thomas Harriot College of Arts and Sciences
    • Biology
    • View Item
    JavaScript is disabled for your browser. Some features of this site may not work without it.

    Browse

    All of The ScholarShipCommunities & CollectionsDateAuthorsTitlesSubjectsTypeDate SubmittedThis CollectionDateAuthorsTitlesSubjectsTypeDate Submitted

    My Account

    Login

    Statistics

    View Google Analytics Statistics

    Effect of Versican G1 Domain Overexpression on Hyaluronan, Hyaluronidase-2, and Synovial Joint Formation in the Embryonic Chick

    Thumbnail
    View/ Open
    Andrews_ecu_0600M_11021.pdf (1.621Mb)

    Show full item record
    Author
    Andrews, Kristen Nichole
    Abstract
    Versican, an extracellular chondroitin sulfate proteoglycan which binds hyaluronan, has been hypothesized to play a role in synovial joint formation in the chick embryo. Several studies have shown versican to be involved in mesenchymal aggregation during limb chondrogenesis with diminished expression as later stages of limb development occur. Although expression within the cartilage model is decreased, versican is still found in a few areas including the joint interzone and perichondrium. The presence of versican along with hyaluronan in the interzone could further joint interzone formation, the process of which is still not completely understood. Previous studies showed that the hyaluronan degratory enzyme, hyaluronidase 2 (Hyal-2), was up-regulated in the developing joint by overexpression of the hyaluronan-binding G1 domain of versican, suggesting a role for versican in regulation of hyaluronan metabolism during early stages of synovial joint morphogenesis. The goal of this study was to manipulate versican and hyaluronan expression in the embryonic chick to obtain a better understanding of their interaction during synovial joint development. To study versican's function, an adenovirally-encoded G1 N-terminal hyaluronan-binding domain was over-expressed via microinjection into the presumptive elbow joint of embryonic chicks. To examine the potential interaction of versican with the hyaluronan pathway during joint development, microinjection of dominant negative CD44 adenovirus, the hyaluronan synthesis inhibitor 4-methylumbelliferone, and two hyaluronidase2 shRNA viruses (hyal2-702 and hyal2-1130) with or without co-injection of adeno-G1 versican was performed. After injection, changes in expression of hyaluronan were observed via histochemistry at early cavitation stages (HH35/36). Morphological changes in the developing joint were observed using whole mount Alcian blue histochemistry and histochemical/immunocytochemical examination of tissue sections. To investigate whether adeno-G1 versican effects were dependent on hyaluronan or CD44, hyaluronan ELISA was performed to assess its accumulation due to up-regulation of Hyal-2. Overexpression of versican G1 in the developing synovial joint led to an enlargement of the interzone area and increased hyaluronan staining was seen in areas of adeno-G1 infection within treated limbs in comparison to controls. Treatment with 4-methylumbelliferone in conjunction with adeno-G1 also resulted in an increased interzone area but an overall decrease in hyaluronan accumulation. Hyaluronan ELISA results demonstrated reduced hyaluronan levels in the 4-methylumbelliferone treated samples in comparison to adeno-G1 alone. Immunohistochemical/histochemical staining along with trends seen from real-time PCR analysis of Hyal-2 suggests that the ability of G1 versican overexpression to upregulate Hyal-2 is dependent on hyaluronan.  
    URI
    http://hdl.handle.net/10342/4189
    Subject
     Biology; Developmental biology; CD44; Hyaluronan; Hyaluronidase-2; Limb development; Versican 
    Date
    2013
    Citation:
    APA:
    Andrews, Kristen Nichole. (January 2013). Effect of Versican G1 Domain Overexpression on Hyaluronan, Hyaluronidase-2, and Synovial Joint Formation in the Embryonic Chick (Master's Thesis, East Carolina University). Retrieved from the Scholarship. (http://hdl.handle.net/10342/4189.)

    Display/Hide MLA, Chicago and APA citation formats.

    MLA:
    Andrews, Kristen Nichole. Effect of Versican G1 Domain Overexpression on Hyaluronan, Hyaluronidase-2, and Synovial Joint Formation in the Embryonic Chick. Master's Thesis. East Carolina University, January 2013. The Scholarship. http://hdl.handle.net/10342/4189. September 21, 2023.
    Chicago:
    Andrews, Kristen Nichole, “Effect of Versican G1 Domain Overexpression on Hyaluronan, Hyaluronidase-2, and Synovial Joint Formation in the Embryonic Chick” (Master's Thesis., East Carolina University, January 2013).
    AMA:
    Andrews, Kristen Nichole. Effect of Versican G1 Domain Overexpression on Hyaluronan, Hyaluronidase-2, and Synovial Joint Formation in the Embryonic Chick [Master's Thesis]. Greenville, NC: East Carolina University; January 2013.
    Collections
    • Biology
    • Master's Theses
    Publisher
    East Carolina University

    xmlui.ArtifactBrowser.ItemViewer.elsevier_entitlement

    East Carolina University has created ScholarShip, a digital archive for the scholarly output of the ECU community.

    • About
    • Contact Us
    • Send Feedback