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      Immunohistochemical examination of the uteroplacental interface of cows on days 21, 31, 40, and 67 of gestation

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          Abstract

          In brief

          Cattle are classified as having synepitheliochorial placentation in which the majority of the uterine luminal epithelial cells remain intact with some luminal epithelial cells fusing with binucleate trophoblast cells to form syncytial trinucleate cells. This study suggests the possibility that, for a limited and as yet undefined period of gestation, the majority of luminal epithelial cells are eliminated and replaced by trophoblast cells that express pregnancy-associated glycoproteins.

          Abstract

          What we understand about the early stages of placentation in cattle is based on an elegant series of electron microscopic images that provide exquisite detail but limited appreciation for the microanatomy across the uteroplacental interface. In order to achieve a global perspective on the histology of bovine placentation during critical early stages of gestation, i.e., days 21, 31, 40, and 67, we performed immunohistochemistry to detect cell-specific expression of pregnancy-associated glycoprotein (PAG), cytokeratin, epithelial (E)-cadherin, and serine hydroxymethyltransferase 2 (SHMT2) at the intact uteroplacental interface. Key findings from the immunohistochemical analyses are that there are: (i) PAG-positive cells with a single nucleus within the uterine luminal epithelial (LE) cells; (ii) PAG-positive cells with two nuclei in the LE; (iii) PAG-positive syncytial cells with more than three nuclei in the LE; (iv) LE cells that are dissociated from one another and from the basement membrane in regions of syncytialization within the LE layer; (v) replacement of the mononuclear LE with a multilayer thick population of PAG-positive cells invading into the uterine stroma of caruncles but not into the stroma of intercaruncular endometrium; and (vi) PAG-, E-cadherin-, and SHMT2-positive mononuclear cells at the leading edge of developing cotyledonary villi that eventually represent the majority of the epithelial surface separating caruncular stroma from cotyledonary stroma. Finally, the uteroplacental interface of ruminants is not always uniform across a single cross section of a site of placentation, which allows different conclusions to be made depending on the part of the uteroplacental interface being examined.

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          Integrins in cell migration.

          Integrin-based adhesion has served as a model for studying the central role of adhesion in migration. In this article, we outline modes of migration, both integrin-dependent and -independent in vitro and in vivo. We next discuss the roles of adhesion contacts as signaling centers and linkages between the ECM and actin that allows adhesions to serve as traction sites. This includes signaling complexes that regulate migration and the interplay among adhesion, signaling, and pliability of the substratum. Finally, we address mechanisms of adhesion assembly and disassembly and the role of adhesion in cellular polarity.
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            The establishment of an ELISA for the detection of pregnancy-associated glycoproteins (PAGs) in the serum of pregnant cows and heifers.

            The pregnancy-associated glycoproteins (PAGs) are a large gene family expressed in trophoblast cells of ruminant ungulates. The detection of PAGs (more specifically, PAG-1) in maternal serum has served as the basis for pregnancy detection in cattle. Unfortunately, PAG-1 and/or antigenically-related PAGs exhibit a long half-life in maternal serum (>8 d) and can be detected 80-100 d post-partum, thereby producing false positives in animals bred within 60-d of calving. The goal of the present studies was to develop a monoclonal-based assay that targeted early-pregnancy PAGs whose persistence in maternal serum post-partum might be relatively short-lived. Three anti-PAG monoclonal antibodies that recognized distinct subsets of PAGs were selected and used as trapping reagents in a 'sandwich' type of enzyme-linked immunosorbant assay (ELISA). A polyclonal antiserum with broad specificity was used for detecting bound PAGs. A total of 42 cows and heifers were bled daily on day 15, days 22 to 28, and then weekly throughout pregnancy and for 10 weeks (approximately 70 d) into the post-partum period. The ELISA was able to detect PAG in maternal serum of all animals unambiguously by day 28 post-insemination (PAG concentration: 8.75 +/- 3.04 ng/mL). In maternal serum, PAG concentrations peaked during the week of parturition at 588.9 +/- 249.9 ng/mL, and after calving, PAG was completely cleared (half-life: 4.3 d) by eight-week post-partum in 38 of 40 of the animals tested and was at very low concentrations in the remaining two (1.4 and 4.9 ng/mL, respectively). In summary, a monoclonal-based assay has been established that is sensitive enough to detect PAG in maternal serum by the forth week of pregnancy, but does not suffer from carry-over of antigen from a previous pregnancy.
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              Circulating concentrations of bovine pregnancy-associated glycoproteins and late embryonic mortality in lactating dairy herds.

              The objectives of these experiments were as follows: (1) to determine the association between circulating concentrations of pregnancy-associated glycoproteins (PAG) and late embryonic mortality (EM) in lactating dairy cattle following fixed-time artificial insemination (TAI) on d 0 or timed embryo transfer (TET) on d 7, (2) to identify a circulating concentration of PAG on d 31 below which late EM would be likely to occur, and (3) to identify when during gestation (d 31-59) late EM is occurring. Cows were diagnosed pregnant on d 31 of gestation based on presence of a fetal heartbeat and reconfirmed to be pregnant on d 59 of gestation. Late EM occurred when a cow had a viable embryo on d 31 of gestation but not on d 59 following TAI or TET. Only pregnant cows on d 31 were included in the analysis (TAI-maintained, n=413; TAI-EM, n=77; TET-maintained, n=238; TET-EM, n=47). Cows that were pregnant at d 31 of gestation and maintained the pregnancy until d 59 had significantly higher circulating concentrations of PAG at d 31 of gestation compared with cows that experienced late EM between d 31 and 59 of gestation in both TAI and TET. To conduct a more stringent test of the effectiveness of a single circulating PAG concentration (d 31) to predict EM, a receiver-operating characteristic curve was generated to identify a PAG concentration on d 31 that would predict EM with ≥95% accuracy in cows that received TAI or TET. Based on positive and negative predicative value analysis, a circulating concentration of PAG below 1.4 ng/mL (TAI; minimal detectable level 0.28 ng/mL) and 1.85 ng/mL (TET) was 95% accurate in predicting EM (between d 31 and 59) at d 31 of gestation, respectively. Following TET, embryonic loss was tracked by Doppler ultrasound, progesterone, and PAG from d 24 to 59 of gestation, with more than 50% of the loss occurring between d 31 and 38 of gestation. In summary, circulating concentrations of PAG on d 31 of gestation may provide a good marker for predicting EM between d 31 and 59 of gestation, and the data suggest that this model could help predict which cows will undergo late EM.
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                Author and article information

                Journal
                Reproduction
                Reproduction
                REP
                Reproduction (Cambridge, England)
                Bioscientifica Ltd (Bristol )
                1470-1626
                1741-7899
                29 January 2024
                19 December 2023
                01 February 2024
                : 167
                : 2
                : e230444
                Affiliations
                [1 ]Department of Veterinary Integrative Biosciences , School of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, Texas, USA,
                [2 ]Department of Animal Science , College of Agriculture and Life Sciences, Texas A&M University, College Station, Texas, USA
                Author notes
                Correspondence should be addressed to K G Pohler or G A Johnson; Email: Ky.Pohler@ 123456ag.tamu.edu or GJohnson@ 123456cvm.tamu.edu
                Author information
                http://orcid.org/0000-0003-1949-1797
                Article
                REP-23-0444
                10.1530/REP-23-0444
                10895283
                38112573
                121101c5-3d68-452f-b7d8-b5384283bdc3
                © the author(s)

                This work is licensed under a Creative Commons Attribution 4.0 International License.

                History
                : 13 November 2023
                : 19 December 2023
                Funding
                Funded by: National Institute of Food and Agriculture, doi http://dx.doi.org/10.13039/100005825;
                Categories
                Research

                Obstetrics & Gynecology
                Obstetrics & Gynecology

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