The Sertoli cell ectoplasmic specialization is a specialized domain of the calcium-dependent Sertoli-spermatid adherens junction. Structurally abnormal or absent Sertoli ectoplasmic specializations are associated with spermatid sloughing and subsequent oligospermia in conditions associated with reduced fertility potential, although the junctional strength between these cells is not known. Adjudin is a potential male contraceptive agent thought to interrupt testicular binding dynamics of adherens junctions, resulting in controlled spermatid sloughing.It was hypothesized that the mechanism of action of Adjudin, pertinent to its putative contraceptive effect, is the disruption of the Sertoli cell-spermatid junction.
This was tested in vitro using primary isolates of germ cells and both primary and immortal Sertoli cells.This dissertation presents the examination of Sertoli-germ cell interactions in three parts, which address the overall aims of this dissertation project: (1) measurement of the junctional strength between Sertoli cells and spermatids in vitro, (2) determination of the efficacy of sk Sertoli cell lines in Sertoli-germ cell binding studies in vitro, and (3) assessment of Adjudin as a potential male contraceptive, by measuring the junctional binding strength between Sertoli cells and spermatids exposed to this chemical in vitro.For the first time, the strength of the Sertoli-spermatid junction has been measured, using a micropipette pressure transducing system (MPTS).
Results reported in this dissertation demonstrate that the junctional strength between Sertoli cells and germ cells can be measured in vitro, support long held speculations regarding Sertoli-spermatid junctional interactions, and provide a technology to test proposed mechanisms of junctional binding dynamics between cells of the seminiferous epithelium (Chapter 2). Although the sk cell lines initially expressed mRNA for the FSH receptor, coculture results determined that these cell lines have limited value for investigating Sertoli-germ cell binding dynamics in vitro (Chapter 3). By utilizing the MPTS and primary cell isolates, Adjudin was determined to reduce the junctional strength between Sertoli cells and step-8 spermatids. In conclusion, results support the use of Adjudin as a potential reversible male contraceptive agent by a mechanism which alters the adhesion properties between the step-8 spermatid and the Sertoli cell (Chapter 4).
| Identifer | oai:union.ndltd.org:USF/oai:scholarcommons.usf.edu:etd-3761 |
| Date | 01 June 2006 |
| Creators | Wolski, Katja Margrit |
| Publisher | Scholar Commons |
| Source Sets | University of South Flordia |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | Graduate Theses and Dissertations |
| Rights | default |
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