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Endometrial mucins and human embryo implantation. / Sean Gereint Griffiths

Swansea University Author: Sean Gereint Griffiths

Abstract

Failure of the embryo to implant into the uterine lining results in infertility and is also the rate limiting step in FVF. The tethered, glycoprotein MUC1 is a protective cell surface receptor that has been associated with infertility. Evidence suggests MUC1 and other endometrial mucins regulate emb...

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Published: 2013
Institution: Swansea University
Degree level: Doctoral
Degree name: Ph.D
URI: https://cronfa.swan.ac.uk/Record/cronfa42530
first_indexed 2018-08-02T18:54:55Z
last_indexed 2019-10-21T16:47:59Z
id cronfa42530
recordtype RisThesis
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The protein backbone of MUC1 is thought to act as a scaffold for L-selectin ligands and may be integral to the initial tethering of the embryo during early implantation. Objectives The remit of this thesis was to model the expression of MUC1 and other mucins in the human endometrium using endometrial cell lines and to use atomic force microscopy to understand the role played by these proteins in shaping the non-specific and embryo specific adhesive characteristics of endometrial monolayers. If possible the proposed L-selectin implantation mechanism was to be identified and functionally characterised. Methodology This project successfully married the traditional molecular tools of quantitative PCR and immunocytochemistry with novel application of INCELL analyzer high content screening protein analysis, peak force quantitative nano-mechanical mapping atomic force microscopy and single molecule force spectroscopy atomic force microscopy to characterise the specific and non-specific surface adhesion on live endometrial monolayers. Results Firstly, immunocytochemistry and qRTPCR revealed that basal MUC1 expression was significantly higher in Hec-1-B relative to Hec-IA, Ishikawa and Hec50. Secondly, INCELL analysis qualified a distinct heterogeneous expression of MUC1 across the endometrial monolayer and delineated altered patterning following treatment with estradiol and progestins. Thirdly, we have shown a direct and proportional correlation between MUC1 expression and adhesion in live Hec-IA and Hec-IB cell monolayers. Fourthly, this work has confirmed and characterised binding of recombinant L-selectin to the endometrial epithelial cell surface. Fifthly, it is shown that L-selectin surface binding decreases following a reduction in MUC1 surface presentation. Conclusions The results implicate MUC1 as a key component of endometrial adhesion and an initial mediator of implantation with a functional patterned expression suggesting areas of altered receptivity exist across endometrial monolayers. Abnormal MUC1 expression has been shown in endometrial pathologies and unexplained infertility. The current investigation suggests MUC1 protein may assist embryo attachment by retarding it sufficiently through mechanical impedance to allow specific L-selectin binding further securing the embryo. A non-receptive endometrium may contribute towards the infertile phenotype despite repeated IVF treatments, thus novel examination of potential embryo adhesion molecules such as MUC1 may aid understanding of endometrial characteristics which prevent embryo implantation and contribute towards IVF failure.</abstract><type>E-Thesis</type><journal/><journalNumber></journalNumber><paginationStart/><paginationEnd/><publisher/><placeOfPublication/><isbnPrint/><issnPrint/><issnElectronic/><keywords>Medicine.;Developmental biology.;Obstetrics.</keywords><publishedDay>31</publishedDay><publishedMonth>12</publishedMonth><publishedYear>2013</publishedYear><publishedDate>2013-12-31</publishedDate><doi/><url/><notes/><college>COLLEGE NANME</college><department>Swansea University Medical School</department><CollegeCode>COLLEGE CODE</CollegeCode><institution>Swansea University</institution><degreelevel>Doctoral</degreelevel><degreename>Ph.D</degreename><apcterm/><lastEdited>2018-08-29T14:40:23.3157184</lastEdited><Created>2018-08-02T16:24:29.5717948</Created><path><level id="1">Faculty of Medicine, Health and Life Sciences</level><level id="2">Swansea University Medical School - Medicine</level></path><authors><author><firstname>Sean Gereint</firstname><surname>Griffiths</surname><orcid>NULL</orcid><order>1</order></author></authors><documents><document><filename>0042530-02082018162501.pdf</filename><originalFilename>10805279.pdf</originalFilename><uploaded>2018-08-02T16:25:01.7570000</uploaded><type>Output</type><contentLength>24836629</contentLength><contentType>application/pdf</contentType><version>E-Thesis</version><cronfaStatus>true</cronfaStatus><embargoDate>2018-08-02T16:25:01.7570000</embargoDate><copyrightCorrect>false</copyrightCorrect></document></documents><OutputDurs/></rfc1807>
spelling 2018-08-29T14:40:23.3157184 v2 42530 2018-08-02 Endometrial mucins and human embryo implantation. 3a8800ed1dc9668a77e3bfaccfd3590c NULL Sean Gereint Griffiths Sean Gereint Griffiths true true 2018-08-02 Failure of the embryo to implant into the uterine lining results in infertility and is also the rate limiting step in FVF. The tethered, glycoprotein MUC1 is a protective cell surface receptor that has been associated with infertility. Evidence suggests MUC1 and other endometrial mucins regulate embryo implantation. Endometrial epithelia must be shielded from infection whilst permitting the recognition and implantation of the embryo. The protein backbone of MUC1 is thought to act as a scaffold for L-selectin ligands and may be integral to the initial tethering of the embryo during early implantation. Objectives The remit of this thesis was to model the expression of MUC1 and other mucins in the human endometrium using endometrial cell lines and to use atomic force microscopy to understand the role played by these proteins in shaping the non-specific and embryo specific adhesive characteristics of endometrial monolayers. If possible the proposed L-selectin implantation mechanism was to be identified and functionally characterised. Methodology This project successfully married the traditional molecular tools of quantitative PCR and immunocytochemistry with novel application of INCELL analyzer high content screening protein analysis, peak force quantitative nano-mechanical mapping atomic force microscopy and single molecule force spectroscopy atomic force microscopy to characterise the specific and non-specific surface adhesion on live endometrial monolayers. Results Firstly, immunocytochemistry and qRTPCR revealed that basal MUC1 expression was significantly higher in Hec-1-B relative to Hec-IA, Ishikawa and Hec50. Secondly, INCELL analysis qualified a distinct heterogeneous expression of MUC1 across the endometrial monolayer and delineated altered patterning following treatment with estradiol and progestins. Thirdly, we have shown a direct and proportional correlation between MUC1 expression and adhesion in live Hec-IA and Hec-IB cell monolayers. Fourthly, this work has confirmed and characterised binding of recombinant L-selectin to the endometrial epithelial cell surface. Fifthly, it is shown that L-selectin surface binding decreases following a reduction in MUC1 surface presentation. Conclusions The results implicate MUC1 as a key component of endometrial adhesion and an initial mediator of implantation with a functional patterned expression suggesting areas of altered receptivity exist across endometrial monolayers. Abnormal MUC1 expression has been shown in endometrial pathologies and unexplained infertility. The current investigation suggests MUC1 protein may assist embryo attachment by retarding it sufficiently through mechanical impedance to allow specific L-selectin binding further securing the embryo. A non-receptive endometrium may contribute towards the infertile phenotype despite repeated IVF treatments, thus novel examination of potential embryo adhesion molecules such as MUC1 may aid understanding of endometrial characteristics which prevent embryo implantation and contribute towards IVF failure. E-Thesis Medicine.;Developmental biology.;Obstetrics. 31 12 2013 2013-12-31 COLLEGE NANME Swansea University Medical School COLLEGE CODE Swansea University Doctoral Ph.D 2018-08-29T14:40:23.3157184 2018-08-02T16:24:29.5717948 Faculty of Medicine, Health and Life Sciences Swansea University Medical School - Medicine Sean Gereint Griffiths NULL 1 0042530-02082018162501.pdf 10805279.pdf 2018-08-02T16:25:01.7570000 Output 24836629 application/pdf E-Thesis true 2018-08-02T16:25:01.7570000 false
title Endometrial mucins and human embryo implantation.
spellingShingle Endometrial mucins and human embryo implantation.
Sean Gereint Griffiths
title_short Endometrial mucins and human embryo implantation.
title_full Endometrial mucins and human embryo implantation.
title_fullStr Endometrial mucins and human embryo implantation.
title_full_unstemmed Endometrial mucins and human embryo implantation.
title_sort Endometrial mucins and human embryo implantation.
author_id_str_mv 3a8800ed1dc9668a77e3bfaccfd3590c
author_id_fullname_str_mv 3a8800ed1dc9668a77e3bfaccfd3590c_***_Sean Gereint Griffiths
author Sean Gereint Griffiths
author2 Sean Gereint Griffiths
format E-Thesis
publishDate 2013
institution Swansea University
college_str Faculty of Medicine, Health and Life Sciences
hierarchytype
hierarchy_top_id facultyofmedicinehealthandlifesciences
hierarchy_top_title Faculty of Medicine, Health and Life Sciences
hierarchy_parent_id facultyofmedicinehealthandlifesciences
hierarchy_parent_title Faculty of Medicine, Health and Life Sciences
department_str Swansea University Medical School - Medicine{{{_:::_}}}Faculty of Medicine, Health and Life Sciences{{{_:::_}}}Swansea University Medical School - Medicine
document_store_str 1
active_str 0
description Failure of the embryo to implant into the uterine lining results in infertility and is also the rate limiting step in FVF. The tethered, glycoprotein MUC1 is a protective cell surface receptor that has been associated with infertility. Evidence suggests MUC1 and other endometrial mucins regulate embryo implantation. Endometrial epithelia must be shielded from infection whilst permitting the recognition and implantation of the embryo. The protein backbone of MUC1 is thought to act as a scaffold for L-selectin ligands and may be integral to the initial tethering of the embryo during early implantation. Objectives The remit of this thesis was to model the expression of MUC1 and other mucins in the human endometrium using endometrial cell lines and to use atomic force microscopy to understand the role played by these proteins in shaping the non-specific and embryo specific adhesive characteristics of endometrial monolayers. If possible the proposed L-selectin implantation mechanism was to be identified and functionally characterised. Methodology This project successfully married the traditional molecular tools of quantitative PCR and immunocytochemistry with novel application of INCELL analyzer high content screening protein analysis, peak force quantitative nano-mechanical mapping atomic force microscopy and single molecule force spectroscopy atomic force microscopy to characterise the specific and non-specific surface adhesion on live endometrial monolayers. Results Firstly, immunocytochemistry and qRTPCR revealed that basal MUC1 expression was significantly higher in Hec-1-B relative to Hec-IA, Ishikawa and Hec50. Secondly, INCELL analysis qualified a distinct heterogeneous expression of MUC1 across the endometrial monolayer and delineated altered patterning following treatment with estradiol and progestins. Thirdly, we have shown a direct and proportional correlation between MUC1 expression and adhesion in live Hec-IA and Hec-IB cell monolayers. Fourthly, this work has confirmed and characterised binding of recombinant L-selectin to the endometrial epithelial cell surface. Fifthly, it is shown that L-selectin surface binding decreases following a reduction in MUC1 surface presentation. Conclusions The results implicate MUC1 as a key component of endometrial adhesion and an initial mediator of implantation with a functional patterned expression suggesting areas of altered receptivity exist across endometrial monolayers. Abnormal MUC1 expression has been shown in endometrial pathologies and unexplained infertility. The current investigation suggests MUC1 protein may assist embryo attachment by retarding it sufficiently through mechanical impedance to allow specific L-selectin binding further securing the embryo. A non-receptive endometrium may contribute towards the infertile phenotype despite repeated IVF treatments, thus novel examination of potential embryo adhesion molecules such as MUC1 may aid understanding of endometrial characteristics which prevent embryo implantation and contribute towards IVF failure.
published_date 2013-12-31T13:32:03Z
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score 11.047588

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