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A WESTERN BLOT ANALYSIS AND IMMUNOHISTOCHEMICAL STUDY
OF 4-DAY OLD MICE TEETH OVER-EXPRESSING AMELOTIN DURING
AMELOGENESIS
by
Desiree Yazdanshenas
A Thesis Presented to the
FACULTY OF THE GRADUATE SCHOOL
UNIVERSITY OF SOUTHERN CALIFORNIA
In Partial Fulfillment of the
Requirements for the Degree
MASTER OF SCIENCE
(CRANIOFACIAL BIOLOGY)
May 2009
Copyright 2009 Desiree Yazdanshenas
Object Description
| Title | A Western blot analysis and immunohistochemical study of 4-day old mice teeth over-expressing amelotin during amelogenesis |
| Author | Yazdanshenas, Desiree |
| Author email | yazdansh@usc.edu; dyazdanshenas@yahoo.com |
| Degree | Master of Science |
| Document type | Thesis |
| Degree program | Cranio-Facial Biology |
| School | School of Dentistry |
| Date defended/completed | 2009-03-18 |
| Date submitted | 2009 |
| Restricted until | Unrestricted |
| Date published | 2009-05-09 |
| Advisor (committee member) |
Paine, Michael Snead, Malcolm Le, Anh |
| Abstract | Amelotin is an enamel-specific gene product, and it’s expression under normal circumstances is limited to maturation-stage (or late-stage) amelogenesis. The altered pattern of amelotin gene expression is achieved by creating transgenic animals in which the amelogenin gene promoter is used to drive amelotin. This study sets out to analyze transgenic animals in which amelotin is expressed throughout all stages of enamel formation (amelogenesis). This thesis will include an introduction and background in tooth development (including amelogenesis), and present results demonstrating animal genotype confirmation, transgene expression profiles by immunohistochemical and Western analysis and enamel phenotype using electron microscopy. We hypothesized that "The transgene will be functional, and that the amelotin transgene will be expressed throughout all stages of amelogenesis, resulting in an abnormal enamel structure." Data from immunohistochemical studies showed amelotin to be expressed throughout all stages of amelogenesis, with relatively higher expression in the maturation stage of amelogenesis. Western blot analysis confirmed the over-expression of amelotin in these animals. Transgene expression was confirmed during all stages of amelogenesis, and scanning electron microscopy (SEM) imaging showed that the transgenic animals had a highly disrupted enamel structure. The conclusions were that transgenic mice over-expressing amelotin during amelogenesis result in a disrupted enamel architecture, and future studies are aimed at defining whether enamel hypomineralization and hypoplasia are also a feature of these teeth. |
| Keyword | amelotin; enamel; enamelin; tooth development; enamel proteins; amelogenesis; enamel development |
| Language | English |
| Part of collection | University of Southern California dissertations and theses |
| Publisher (of the original version) | University of Southern California |
| Place of publication (of the original version) | Los Angeles, California |
| Publisher (of the digital version) | University of Southern California. Libraries |
| Provenance | Electronically uploaded by the author |
| Type | texts |
| Legacy record ID | usctheses-m2052 |
| Rights | Yazdanshenas, Desiree |
| Repository name | Libraries, University of Southern California |
| Repository address | Los Angeles, California |
| Repository email | http://www.usc.edu/isd/libraries/services/ask_a_librarian/email/ |
| Filename | etd-Yazdanshenas-2782 |
| Archival file | uscthesesreloadpub_Volume29/etd-Yazdanshenas-2782.pdf |
Description
| Title | Page 1 |
| Full text | ! A WESTERN BLOT ANALYSIS AND IMMUNOHISTOCHEMICAL STUDY OF 4-DAY OLD MICE TEETH OVER-EXPRESSING AMELOTIN DURING AMELOGENESIS by Desiree Yazdanshenas A Thesis Presented to the FACULTY OF THE GRADUATE SCHOOL UNIVERSITY OF SOUTHERN CALIFORNIA In Partial Fulfillment of the Requirements for the Degree MASTER OF SCIENCE (CRANIOFACIAL BIOLOGY) May 2009 Copyright 2009 Desiree Yazdanshenas |
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