The development and characterization of a polarized in vitro model of porcine uterine epithelial cells /
A porcine uterine epithelial cell (pUE) culture system that retains structural and functional properties of the surface epithelium in vivo was developed. Epithelial cells became polarized in vitro and compared favorably to uterine epithelial cells in situ once a transepithelial resistance of >60...
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| Format: | Thesis Book |
| Language: | English |
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[Place of publication not identified] :
[publisher not identified] ;
1996.
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| Subjects: | |
| Online Access: | http://proxy.library.tamu.edu/login?url=http://proquest.umi.com/pqdweb?did=743273371&sid=1&Fmt=2&clientId=2945&RQT=309&VName=PQD |
| Summary: | A porcine uterine epithelial cell (pUE) culture system that retains structural and functional properties of the surface epithelium in vivo was developed. Epithelial cells became polarized in vitro and compared favorably to uterine epithelial cells in situ once a transepithelial resistance of >600 QCM2 was established. Microscopic analysis confirmed the presence of a simple columnar epithelium. Indirect immunofluorescence of the tight junction-associated protein, ZO-1, on the polarized cells indicated the formation of tight junctional complexes. Functional polarity of epithelial cultures was also verified by electrical resistance measurements, basal preference for the secretion of Prostaglandins, apical preference for the release of 35S- methionine incorporated secretary proteins, and apically and basally distinct secretary protein profiles. The polarized pUE model was utilized to study specific molecules (including specific integrin subunits, Muc-1 and specific connexins) believed to be involved in the transition of the pre-receptive to the receptive stage of the uterus. The expression of integrin subunits, extracellular matrix proteins, Muc-1 and connexins was first examined in situ on both uterine epithelium and trophectoderm. Additionally, gilts were ovariectomized and given exogenous steroid treatments (vehicle, estrogen, progesterone, and estrogen plus progesterone). Results from this initial study suggested that progesterone upregulates integrin subunits a4, a5, and 01. lntegrin subunits a,, and P3 were constitutively expressed at a high level. These five integrin subunits were detected on trophectoderm from Day 1 1 -1 5 conceptuses. Additionally, extracellular matrix proteins present on contacting surfaces pUE and trophectoderm (fibronectin and vitronectin) are potential ligands for integrin subunits. Muc-1 was expressed at a moderate intensity in estrus and ovariectomized-estrogen-treated gilts. Although a gap junction-like structure was seen during Days 11 -1 5 of cyclic and pregnant gilts, subsequent studies using immunocytochemistry and a dye transfer assay failed to confirm the presence of connexins. The polarized pUE cells responded to steroid treatment (estrogen, progesterone, estrogen plus progesterone) by reproducing steroid induced changes observed in vivo, including upregulation of integrin subunits a4, a5, and 01 by progesterone, regulation of Muc-1 and the modulation of secreted proteins by steroids. The polarized pUE culture system may be useful for in vitro analysis of maternal recognition of pregnancy paradigms. |
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| Item Description: | Vita. "Major Subject: Veterinary Anatomy". |
| Physical Description: | xiii, 178 leaves : illustrations ; 28 cm. Issued also on microfiche from University Microfilms Inc. |
| Bibliography: | Includes bibliographical references. |