Repository logo
 

Characterization of periattachment factor

Simple item page
dc.contributor.authorPurcell, Scott H., author
dc.contributor.authorAnthony, Russell V., advisor
dc.contributor.authorSeidel, George E., advisor
dc.date.accessioned2024-03-13T20:12:30Z
dc.date.available2024-03-13T20:12:30Z
dc.date.issued2008
dc.description.abstractThe purpose of these studies was to characterize expression of periattachment factor (PF) mRNA in the developing sheep conceptus, localize PF in the sheep conceptus, determine if degrading PF mRNA in the sheep conceptus was detrimental to development, and characterize PF in human tissues and cells. Sheep conceptuses were collected on d 11, 13, 15, 16, 17, 21, and 30 post-mating. oPF mRNA exhibited a quadratic expression pattern (P<0.05). No oPF mRNA was detected in d 11 conceptuses. From d 13, oPF mRNA increased to a peak at d 16 before declining. Peak expression of oPF mRNA in the conceptus occurs during rapid elongation and initial attachment of the conceptus to the endometrium. Immunolocalization of PF in a d 15 conceptus showed predominantly nuclear staining in trophectoderm and trophendoderm. Next, embryos collected from superovulated ewes on d 8 post-mating were infected with either a lentivirus expressing a short-hairpin (sh) RNA designed to target PF mRNA for degradation, a lentivirus expressing a shRNA containing 3 mismatched nucleotides, or a control lentivirus expressing no shRNA. Following infection, blastocysts were transferred into recipient ewes and collected back on d 15 of gestation. While 94 and 88% of the control and mismatched shRNA-treated conceptuses elongated by d 15, none of the embryos treated with the lentivirus expressing shRNA against PF mRNA elongated, and most died. This indicates that oPF is required for conceptus elongation and survival. Human PF mRNA was detected in human carcinomas and in the first trimester cytotrophoblast cell line, hTR8. Immunohistochemistry showed PF in the nuclei of carcinomas and in first and second trimester cytotrophoblasts. PF was also present in the invading cytotrophoblast columns. In an in vitro invasion assay of first trimester cytotrophoblasts, hPF mRNA increased from 0, 3, to 12 h as invasion occurred. To further characterize PF in human cells, a lentiviral construct expressing an shRNA targeting the hPF mRNA sequence was developed that resulted in 63% mRNA reduction in BeWo cells.
dc.format.mediumborn digital
dc.format.mediumdoctoral dissertations
dc.identifierETDF_Purcell_2008_3332711.pdf
dc.identifier.urihttps://hdl.handle.net/10217/237913
dc.languageEnglish
dc.language.isoeng
dc.publisherColorado State University. Libraries
dc.relation.ispartof2000-2019
dc.rightsCopyright and other restrictions may apply. User is responsible for compliance with all applicable laws. For information about copyright law, please see https://libguides.colostate.edu/copyright.
dc.rights.licensePer the terms of a contractual agreement, all use of this item is limited to the non-commercial use of Colorado State University and its authorized users.
dc.subjectimplantation
dc.subjectperiattachment factor
dc.subjecttrophoblasts
dc.subjectphysiology
dc.titleCharacterization of periattachment factor
dc.typeText
dcterms.rights.dplaThis Item is protected by copyright and/or related rights (https://rightsstatements.org/vocab/InC/1.0/). You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s).
thesis.degree.disciplineBiomedical Sciences
thesis.degree.grantorColorado State University
thesis.degree.levelDoctoral
thesis.degree.nameDoctor of Philosophy (Ph.D.)

Files

Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
ETDF_Purcell_2008_3332711.pdf
Size:
1.27 MB
Format:
Adobe Portable Document Format
Download

Collections

2000-2019
Theses and Dissertations