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  • 1
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    Online Resource
    Extracellular matrix and the implantation cascade in pigs
    Bioscientifica ; 2020
    In:  Bioscientifica Proceedings ( 2020-10-12)
    Details
    In: Bioscientifica Proceedings, Bioscientifica, ( 2020-10-12)
    Type of Medium: Online Resource
    ISSN: 2515-0227
    URL: Article
    DOI: 10.1530/biosciprocs.15.0011
    Language: Unknown
    Publisher: Bioscientifica
    Publication Date: 2020
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  • 2
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    Enhanced focal adhesion assembly reflects increased mechanosensation and mechanotransduction at maternal–conceptus interface and uterine wall during ovine pregnancy
    Bioscientifica ; 2009
    In:  REPRODUCTION Vol. 137, No. 3 ( 2009-03), p. 567-582
    Details
    In: REPRODUCTION, Bioscientifica, Vol. 137, No. 3 ( 2009-03), p. 567-582
    Abstract: The integrity of the fetal–maternal interface is critical for proper fetal nourishment during pregnancy. Integrins are important adhesion molecules present at the interface during implantation; however, in vivo evidence for integrin activation and focal adhesion formation at the maternal–conceptus interface is limited. We hypothesized that focal adhesion assembly in uterine luminal epithelium (LE) and conceptus trophectoderm (Tr) results from integrin binding of extracellular matrix (ECM) at this interface to provide increased tensile forces and signaling to coordinate utero-placental development. An ovine model of unilateral pregnancy was used to evaluate mechanotransduction events leading to focal adhesion assembly at the maternal–conceptus interface and within the uterine wall. Animals were hysterectomized on days 40, 80, or 120 of pregnancy, and uteri immunostained for integrins (ITGAV, ITGA4, ITGA5, ITGB1, ITGB3, and ITGB5), ECM proteins (SPP1, LGALS15, fibronectin (FN), and vitronectin (VTN)), cytoskeletal molecules (ACTN and TLN1), and a signal generator (PTK2). Focal adhesion assembly in myometrium and stroma was also studied to provide a frame of reference for mechanical stretch of the uterine wall. Large focal adhesions containing aggregates of ITGAV, ITGA4, ITGA5, ITGB1, ITGB5, ACTN, and PTK2 were detected in interplacentomal uterine LE and Tr of gravid but not non-gravid uterine horns and increased during pregnancy. SPP1 and LGALS15, but not FN or VTN, were present along LE and Tr interfaces in both uterine horns. These data support the idea that focal adhesion assembly at the maternal–conceptus interface reflects adaptation to increasing forces caused by the growing fetus. Cooperative binding of multiple integrins to SPP1 deposited at the maternal–conceptus interface forms an adhesive mosaic to maintain a tight connection between uterine and placental surfaces along regions of epitheliochorial placentation in sheep.
    Type of Medium: Online Resource
    ISSN: 1470-1626 , 1741-7899
    URL: Article
    DOI: 10.1530/REP-08-0304
    Language: Unknown
    Publisher: Bioscientifica
    Publication Date: 2009
    detail.hit.zdb_id: 2037813-0
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  • 3
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    Glycosylation dependent cell adhesion molecule 1-like protein and l-selectin expression in sheep interplacentomal and placentomal endometrium
    Bioscientifica ; 2006
    In:  Reproduction Vol. 131, No. 4 ( 2006-04), p. 751-761
    Details
    In: Reproduction, Bioscientifica, Vol. 131, No. 4 ( 2006-04), p. 751-761
    Abstract: Glycosylation dependent cell adhesion molecule 1 (GlyCAM-1), a mucin component of sheep histotroph produced by glandular epithelium (GE) during early pregnancy, is hypothesized to function in implantation. However, GlyCAM-1 is present in uterine tissues subsequent to implantation suggesting additional functions of this l -selectin-binding ligand. This study focused on uterine GlyCAM-1 expression during placentome development in sheep. Western blot analysis of day 50 pregnant sheep identified 45, 40, and 25 kDa bands in interplacentomal endometrium, 40 and 25 kDa bands in placentomes, and 80 and 40 kDa bands in chorioallantois. The GlyCAM-1 proteins in interplacentomal regions were comparable to those detected in day 15–19 pregnant sheep, however, the 80 kDa form was unique to chorioallantois, and the absence of the 45 kDa GlyCAM-1 in placentomes indicated differences between interplacentomal and placentomal endometrium. Immunofluorescence identified GlyCAM-1 in lumenal epithelium (LE), stromal fibroblasts, and vascular smooth muscle cells. To better define its cellular distribution, GlyCAM-1 was co-localized with either epithelium-specific cytokeratin, smooth muscle-specific alpha-smooth muscle actin (α SMA), or stromal-specific vimentin. In interplacentomal endometrium, GlyCAM-1 co-localized with cytokeratin in LE but not in GE. GlyCAM-1 did not co-localize with α SMA, and was localized in the extracellular matrix of vimentin-positive stroma. In placentomes, GlyCAM-1 did not co-localize with cytokeratin, but did co-localize with α SMA and vimentin. Thus, in contrast to interplacentomal regions, GlyCAM-1 in placentomes was predominantly localized in vasculature rather than epithelial cells. Further, leukocytes expressing L-selectin were localized to the endothelial surface of GlyCAM-1-expressing vessels within placentomes. These data suggest that GlyCAM-1 assumes distinct functions in compartment-specific regions of the sheep uterus.
    Type of Medium: Online Resource
    ISSN: 1470-1626 , 1741-7899
    URL: Article
    DOI: 10.1530/rep.1.00855
    Language: Unknown
    Publisher: Bioscientifica
    Publication Date: 2006
    detail.hit.zdb_id: 2037813-0
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  • 4
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    Functional analysis of autocrine and paracrine signalling at the uterine–conceptus interface in pigs
    Bioscientifica ; 2020
    In:  Bioscientifica Proceedings ( 2020-10-12)
    Details
    In: Bioscientifica Proceedings, Bioscientifica, ( 2020-10-12)
    Type of Medium: Online Resource
    ISSN: 2515-0227
    URL: Article
    DOI: 10.1530/biosciprocs.16.0014
    Language: Unknown
    Publisher: Bioscientifica
    Publication Date: 2020
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  • 5
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    ITGAV (alpha v integrins) bind SPP1 (osteopontin) to support trophoblast cell adhesion
    Bioscientifica ; 2017
    In:  Reproduction Vol. 153, No. 5 ( 2017-05), p. 695-706
    Details
    In: Reproduction, Bioscientifica, Vol. 153, No. 5 ( 2017-05), p. 695-706
    Abstract: Attachment of the conceptus trophoblast (Tr) to the uterine luminal epithelium (LE) is critical for successful implantation. This study determined whether alpha v (av) integrins (ITGAV) directly mediate porcine trophoblast cell adhesion to secreted phosphoprotein 1 (SPP1, also known as osteopontin (OPN)) and examined the temporal/spatial expression of ITGAV, beta 3 (b3, ITGB3) and beta 6 (b6, ITGB6) integrin subunits, and SPP1, at the uterine–placental interface of pigs. Knockdown of ITGAV in porcine Tr (pTr2) cells by siRNA reduced pTr2 attachment to SPP1. In situ hybridization confirmed the presence of ITGAV , ITGB3 and ITGB6 mRNAs in uterine LE and conceptus Tr between Days 9 and 60 of gestation, with no change in the magnitude of expression over the course of pregnancy. Exogenous E2 or P4 did not affect ITGAV , ITGB3 and ITGB6 mRNA expression in the uteri of ovariectomized gilts. Immunofluorescence identified ITGAV, ITGB3 and SPP1 proteins in large aggregates at the uterine LE-placental Tr/chorion interface on Day 25, but aggregates were no longer observed by Day 50 of gestation. These results are the first to directly demonstrate that pTr2 cells engage ITGAV-containing integrin receptors to adhere to SPP1 and suggest that mechanical forces generated by tethering elongating conceptuses to uterine LE leads to assembly of focal adhesions containing ITGAV and SPP1; however, as placentation progresses, subsequent folding/interdigitation at the uterine–placental interface disperses mechanical forces resulting in the loss of focal adhesions.
    Type of Medium: Online Resource
    ISSN: 1470-1626 , 1741-7899
    URL: Article
    DOI: 10.1530/REP-17-0043
    Language: Unknown
    Publisher: Bioscientifica
    Publication Date: 2017
    detail.hit.zdb_id: 2037813-0
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  • 6
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    Secreted phosphoprotein 1 (osteopontin) is expressed by stromal macrophages in cyclic and pregnant endometrium of mice, but is induced by estrogen in luminal epithelium during conceptus attachment for implantation
    Bioscientifica ; 2006
    In:  Reproduction Vol. 132, No. 6 ( 2006-12), p. 919-929
    Details
    In: Reproduction, Bioscientifica, Vol. 132, No. 6 ( 2006-12), p. 919-929
    Abstract: Secreted phosphoprotein 1 (SPP1, osteopontin) is the most highly upregulated extracellular matrix/adhesion molecule/cytokine in the receptive phase human uterus, and Spp1 null mice manifest decreased pregnancy rates during mid-gestation as compared with wild-type counterparts. We hypothesize that Spp1 is required for proliferation, migration, survival, adhesion, and remodeling of cells at the conceptus–maternal interface. Our objective was to define the temporal/spatial distribution and steroid regulation of Spp1 in mouse uterus during estrous cycle and early gestation. In situ hybridization localized Spp1 to luminal epithelium (LE) and immune cells. LE expression was prominent at proestrus, decreased by estrus, and was nearly undetectable at diestrus. During pregnancy, Spp1 mRNA was not detected in LE until day 4.5 (day 1 = vaginal plug). Spp1 -expressing immune cells were scattered within the endometrial stroma throughout the estrous cycle and early pregnancy. Immunoreactive Spp1 was prominent at the apical LE surface by day 4.5 of pregnancy and Spp1 protein was also co-localized with subsets of CD45-positive (leukocytes) and F4/80-positive (macrophages) cells. In ovariectomized mice, estrogen, but not progesterone, induced Spp1 mRNA, whereas estrogen plus progesterone did not induce Spp1 in LE. These results establish that estrogen regulates Spp1 in mouse LE and are the first to identify macrophages that produce Spp1 within the peri-implantation endometrium of any species. We suggest that Spp1 at the apical surface of LE provides a mechanism to bridge conceptus to LE during implantation, and that Spp1-positive macrophages within the stroma may be involved in uterine remodeling for conceptus invasion.
    Type of Medium: Online Resource
    ISSN: 1470-1626 , 1741-7899
    URL: Article
    DOI: 10.1530/REP-06-0068
    Language: Unknown
    Publisher: Bioscientifica
    Publication Date: 2006
    detail.hit.zdb_id: 2037813-0
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  • 7
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    Online Resource
    OPN binds alpha V integrin to promote endothelial progenitor cell incorporation into vasculature
    Bioscientifica ; 2020
    In:  Reproduction Vol. 159, No. 4 ( 2020-04), p. 465-478
    Details
    In: Reproduction, Bioscientifica, Vol. 159, No. 4 ( 2020-04), p. 465-478
    Abstract: Angiogenesis is fundamental to the expansion of the placental vasculature during pregnancy. Integrins are associated with vascular formation; and osteopontin is a candidate ligand for integrins to promote angiogenesis. Endothelial progenitor cells (EPCs) are released from bone marrow into the blood and incorporate into newly vascularized tissue where they differentiate into mature endothelium. Results of studies in women suggest that EPCs may play an important role in maintaining placental vascular integrity during pregnancy, although little is known about how EPCs are recruited to these tissues. Our goal was to determine the αv integrin mediated effects of osteopontin on EPC adhesion and incorporation into angiogenic vascular networks. EPCs were isolated from 6 h old piglets. RT-PCR revealed that EPCs initially had a monocyte-like phenotype in culture that became more endothelial-like with cell passage. Immunofluorescence microscopy confirmed that the EPCs express platelet endothelial cell adhesion molecule, vascular endothelial cadherin, and von Willebrand factor. When EPCs were cultured on OPN-coated slides, the αv integrin subunit was observed in focal adhesions at the basal surface of EPCs. Silencing of αv integrin reduced EPC binding to OPN and focal adhesion assembly. In vitro siRNA knockdown in EPCs,demonstrated that OPN stimulates EPC incorporation into human umbilical vein endothelial cell (HUVEC) networks via αv-containing integrins. Finally, in situ hybridization and immunohistochemistry localized osteopontin near placental blood vessels. In summary, OPN binds the αv integrin subunit on EPCs to support EPC adhesion and increase EPC incorporation into angiogenic vascular networks.
    Type of Medium: Online Resource
    ISSN: 1470-1626 , 1741-7899
    URL: Article
    DOI: 10.1530/REP-19-0358
    Language: Unknown
    Publisher: Bioscientifica
    Publication Date: 2020
    detail.hit.zdb_id: 2037813-0
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  • 8
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    Online Resource
    Integrin α is necessary for the attachment of trophectoderm cells to osteopontin to mediate adhesion of the conceptus to the uterus during implantation in pigs
    Bioscientifica ; 2019
    In:  Bioscientifica Proceedings ( 2019-04-05)
    Details
    In: Bioscientifica Proceedings, Bioscientifica, ( 2019-04-05)
    Type of Medium: Online Resource
    ISSN: 2515-0227
    URL: Article
    DOI: 10.1530/biosciprocs.19.0020
    Language: Unknown
    Publisher: Bioscientifica
    Publication Date: 2019
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