Identification of embryo implantation-related proteins Mitra Arianmanesh Embryo implantation is a complex process involving an active dialogue between the endometrium and embryo. Tightly controlled communication between the hypothalamus, pituitary, corpus luteum (CL), endometrium and embryo is essential for implantation. Unravelling molecules involved in embryo implantation is essential since implantation failure is one of the main causes of female infertility. Therefore, identification of molecular events during embryo implantation may result in enhancing implantation rates in both natural and assisted reproductive cycles, improving contraceptive design and reducing the rate of multiple pregnancies following embryo transfer in IVF cycles. Thus, in this study, sheep was used as an animal model in order to study endometrial, corpus luteal and plasma proteome changes during embryo implantation and early pregnancy. Endometrium, CLs and plasma were harvested from cyclic ewes on days 12 and 16 of the oestrous cycle (n=4 ewes/group) and from pregnant ewes on days 12, 16 and 20 of pregnancy (n=4 ewes/group). Furthermore, ovine endometrium were collected from pregnant and non-pregnant horns on days 16 (n=4) and 20 (n=4) of pregnancy to compare endometrial protein profiles of the gravid horn (in the presence of the conceptus) with the non-gravid horn (in the absence of the conceptus) in response to the conceptus to elucidate local embryo-endometrial signalling. 2DE gel, LC-MS/MS, Western blot, IHC and qRT-PCR were employed to quantify implantation processes. This study has identified proteins in the CL and endometrium with involvement in biological pathways that are fundamental for embryo implantation and gestation. In addition, it was found that the implanting embryo is capable of regulating the expression of endometrial proteins to establish an ideal environment for its implantation and establishment of pregnancy. These findings provide an addition to the field and a solid base for targeted studies to improve our understanding of implantation and its regulation.

Thesis (M.S.)--California Polytechnic State University, 2009. / Title from PDF title page; viewed on February 2, 2010. Major professor: Matthew A. Burd, D.V.M. "Presented to the faculty of California Polytechnic State University, San Luis Obispo." "In partial fulfillment of the requirements for the degree [of] Master of Science in Agriculture." "December 2009." Includes bibliographical references (p. 47-52).

published_or_final_version / Obstetrics and Gynaecology / Doctoral / Doctor of Philosophy

Implantation is a critical step in reproduction. It is complicated and well-coordinated consisting of apposition, attachment and invasion of embryo into the endometrium. The mechanism of implantation is unclear. Our previous proteomic study showed an increase of annexin A2 in the endometrium during the implantation window of mice, consistent with the increased annexin A2 expression in the receptive human endometrium. The hypothesis of this project was that annexin A2 mediatedthe embryo-endometrium attachment. The first objective was to study the spatio-temporal expression of endometrial annexin A2 immunoreactivities in humans and mice. The cyclical change in annexin A2 expression in the mouse and human reproductive cycle suggested the involvement of a steroid regulatory mechanism. Interestingly, annexinA2 was transiently expressed on the membrane between the mouse uterine luminal epithelium and the implanting embryos from Day 4 (pre-implantation) to Day 5 (post-implantation) of pregnancy. No such signal change was observed at the inter-implantation sites, showing that the implanting embryos partially regulated annexin A2 expression. These observations and the high expression of the molecule in the luminal epithelium of human endometrium in the mid-and late luteal phase were consistent with a role of annexin A2 in implantation. The second objective was to verify the action of steroids on annexin A2 expression. It was found that a combination of 6675 pmol/L of estrogen and 429.8nmol/L of progesterone increased the total and apical surface expression of annexin A2. In mice, estrogen but not progesterone, increased annexin A2 expression in the uterine luminal epithelium of ovariectomized mice. The third objective was to study the function of annexin A2 in embryo-endometrium attachment using an Ishikawa (endometrial epithelial cells)-JEG-3 trophoblast spheroids (embryo surrogate) coculture model. Knockdown of the expression of annexin A2 in either or both cell lines significantly decreased the attachment rate of the spheroids onto the endometrial cells. The suppressive action on the two cell lines was additive. The attachment was also suppressed in the presence of anti-annexin A2 antibody during coculture. Annexin A2 was also involved in mouse implantation as demonstrated by a significant decrease in implantation sites after injection of anti-annexin A2 antibody into the mouse uterine horn. The final objective was to study the action of annexin A2 as an adhesive molecule in embryo attachment. It was found that loss of P11, the binding partner of annexin A2, reduced the attachment rate of the JEG-3 spheroids probably by decreasing the translocation of annexin A2 to the surface of the endometrial cells. Recombinant P11 and annexin A2 protein failed to bind significantly to the Ishikawa cells and the JEG-3 cells. In summary, this study demonstrates the involvement of annexin A2 as an adherent molecule in the embryo-endometrium interaction. / published_or_final_version / Obstetrics and Gynaecology / Doctoral / Doctor of Philosophy

Thesis (M.S.)--Brigham Young University. Dept. of Plant and Animal Sciences, 2004. / Includes bibliographical references.

"December 2003" / Includes bibliographical references (leaves 183-224) / xvii, 236 leaves : ill. (chiefly col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, Dept. of Obstetrics and Gynaecology, 2004

published_or_final_version / Medical Sciences / Master / Master of Medical Sciences

Thesis (Ph.D.)--University of Adelaide, Dept. of Obstetrics and Gynaecology, 2004. / "December 2003" Includes bibliographical references (leaves 183-224).

No description available.

Hemolymph of Tipula trivittata larvae permits freeze tolerance of this insect due to its content of cryoprotectants and ice nucleating proteins. Spontaneous ice nucleation of the dialyzed hemolymph occurs between -5 C and -11 C up to dilutions of 1:1000. The objectives of this study were to evaluate the effects of seeding temperature (-5 C vs -7 C), and the presence of hemolymph at a low (.1% v/v) and a high (10% v/v) level on the survival of frozen-thawed bovine embryos. In Exp. l, survival rates of 6 and 7 day bovine embryos frozen in medium containing .1% hemolymph and seeded at -5 C or -7 C, were compared to evaluate the effect of seeding temperature. The effect of hemolymph was evaluated by including a control without hemolymph seeded at -7 C. In Exp. 2, survival rates of embryos frozen with and without 10% hemolymph were compared. In Exp. 3 the evaluation of the effect of 10% hemolymph was continued. Also included was a control handled identically to embryos frozen in medium with 10% hemolymph regarding pre and post freeze manipulations but which was not frozen. This allowed evaluation of freezing damage per se. For Exps. l, 2, and 3, survival based on mean final development score and time to advance a developmental stage in vitro did not differ for embryos frozen. However, in Exp.· 3, the control which was not frozen had 30% greater survival than embryos undergoing the same manipulations but which were frozen. Thus, neither seeding temperature nor inclusion of .1% or 10% hemolymph in freezing medium had a significant effect on survival of frozen-thawed bovine embryos. / Master of Science