Immunolocalisation of the uterine secretory proteins uterocalin, uteroferrin and uteroglobin in the mare's uterus and placenta throughout pregnancy

Ellenberger, C.; Wilsher, S.; Allen, W. R.; Hoffmann, Christine; Kölling, Malte; Bazer, Fuller W.; Klug, Jörg; Schoon, D.; Schoon, H-A

doi:10.1016/j.theriogenology.2008.04.050

Cited by 43 publications

(36 citation statements)

References 35 publications

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“…Immunoreactive CTSB protein was detected using the rabbit anti-rat CTSB polyclonal IgG (2.5 mg/ml, Catalog number 06-480; Upstate, Lake Placid, NY, USA). Immunoreactive ACP5 protein was detected using affinity-purified rabbit anti-ACP5 (2.5 mg/ml; Ellenberger et al 2008). Multiple exposures of each western blot were performed to ensure linearity of chemiluminescent signals.…”

Section: Western Blot Analysesmentioning

confidence: 99%

Effects of long-term progesterone on developmental and functional aspects of porcine uterine epithelia and vasculature: progesterone alone does not support development of uterine glands comparable to that of pregnancy

Bailey¹,

Dunlap²,

Frank³

et al. 2010

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In pigs, endometrial functions are regulated primarily by progesterone and placental factors including estrogen. Progesterone levels are high throughout pregnancy to stimulate and maintain secretion of histotroph from uterine epithelia necessary for growth, implantation, placentation, and development of the conceptus (embryo and its extra-embryonic membranes). This study determined effects of longterm progesterone on development and histoarchitecture of endometrial luminal epithelium (LE), glandular epithelium (GE), and vasculature in pigs. Pigs were ovariectomized during diestrus (day 12), and then received daily injections of either corn oil or progesterone for 28 days. Prolonged progesterone treatment resulted in increased weight and length of the uterine horns, and thickness of the endometrium and myometrium. Hyperplasia and hypertrophy of GE were not evident, but LE cell height increased, suggesting elevated secretory activity. Although GE development was deficient, progesterone supported increased endometrial angiogenesis comparable to that of pregnancy. Progesterone also supported alterations to the apical and basolateral domains of LE and GE. Dolichos biflorus agglutinin lectin binding and a v integrin were downregulated at the apical surfaces of LE and GE. Claudin-4, a 2 b 1 integrin, and vimentin were increased at basolateral surfaces, whereas occludins-1 and -2, claudin-3, and E-cadherin were unaffected by progesterone treatment indicating structurally competent trans-epithelial adhesion and tight junctional complexes. Collectively, the results suggest that progesterone affects LE, GE, and vascular development and histoarchitecture, but in the absence of ovarian or placental factors, it does not support development of GE comparable to pregnancy. Furthermore, LE and vascular development are highly responsive to the effects of progesterone.

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Section: Western Blot Analysesmentioning

confidence: 99%

Effects of long-term progesterone on developmental and functional aspects of porcine uterine epithelia and vasculature: progesterone alone does not support development of uterine glands comparable to that of pregnancy

Bailey¹,

Dunlap²,

Frank³

et al. 2010

Reproduction

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“…Proteins were denatured and separated by 12% SDS-PAGE, and western blot analysis was performed as described previously (Johnson et al 1999b) using ECL detection (SuperSignal West Pico, Pierce, Rockford, IL, USA) and X-OMAT AR X-ray film (Kodak). Immunoreactive ACP5 protein was detected using affinity purified rabbit anti-ACP5 (2.5 mg/ml; Ellenberger et al 2008) and SPP1 protein was detected using a cocktail containing rabbit polyclonal antibodies against recombinant human SPP1 (anti-hSPP1; LF-123 and LF-124; 5 mg/ml (Fisher et al 1995). Rabbit IgG (5 mg/ml; Sigma) was used as the primary antibody for controls.…”

Section: Western Blot Analysesmentioning

confidence: 99%

“…Immunocytochemical localization of immunoreactive ACP5 in the porcine uterus was performed as described previously (Joyce et al 2005b) in uterine tissue cross sections using rabbit anti-porcine ACP5 (4.0 mg/ml; Ellenberger et al 2008). Antigen retrieval for ACP5 was performed by boiling citrate buffer.…”

Section: Immunohistochemical Analysismentioning

confidence: 99%

Effects of long-term progesterone exposure on porcine uterine gene expression: progesterone alone does not induce secreted phosphoprotein 1 (osteopontin) in glandular epithelium

et al. 2010

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Pigs experience significant conceptus loss near mid-gestation, correlating with increasing glandular epithelial (GE) development and secretory activity. Secreted phosphoprotein 1 (SPP1, osteopontin) increases in GE between days 30 and 40 of pregnancy and is expressed in the GE of day 90 pseudopregnant pigs, suggesting that progesterone (P 4 ) from corpora lutea is responsible for induction of SPP1 in GE. In this study, pigs were ovariectomized and treated daily with P 4 to assess effects of 40 days of P 4 exposure on SPP1, P 4 receptor (PGR), uteroferrin (ACP5), and fibroblast growth factor 7 (FGF7) expression in porcine endometria. PGR mRNA decreased in pigs injected with P 4 compared with pigs injected with corn oil (CO), and PGRs were downregulated in the luminal epithelium (LE) and GE. ACP5 mRNA increased in pigs injected with P 4 compared with pigs injected with CO, and ACP5 was induced in the GE of P 4 -treated pigs. FGF7 mRNA increased in pigs injected with P 4 compared with pigs injected with CO, and FGF7 was induced in the LE and GE of P 4 -treated pigs. SPP1 mRNA was not different between pigs injected with P 4 compared with pigs injected with CO, and SPP1 was not present in the GE of P 4 -treated pigs. Therefore, long-term P 4 , in the absence of ovarian and/or conceptus factors, does not induce SPP1 expression in GE. We hypothesize that a servomechanism involving sequential effects of multiple hormones and cytokines, similar to those for sheep and humans, is required for GE differentiation and function, including the synthesis and secretion of SPP1.

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“…In the mare, there is a distinct uterotubal papilla which presumably helps to maintain the marked differences in fluid composition observed in different parts of the oviduct and uterus in both horses and other domestic and laboratory species; this is in contrast to the lack of an anatomically distinct uterotubal separation in women due to which the oviductal and uterine fluids appear to mix freely (Hunter 1998). During the initial intrauterine period, the equine embryo migrates continually, surrounded by a protective glycoprotein capsule and bathed in nourishing endometrial secretions which contain several progesteronedependent proteins (Ellenberger et al 2008). The addition of one of the major progesterone dominated proteins, uterocalin, to embryo culture medium had a positive effect on capsule formation around IVP blastocysts.…”

Section: Development Of Horse Embryosmentioning

confidence: 99%

“…As a result of its molecular structure, uterocalin has since been proposed to function primarily as a carrier of biologically important lipids and a source of essential amino acids for the developing conceptus (Suire et al 2001, Kennedy 2004, where the positive charge of uterocalin (Crossett et al 1996) is thought to facilitate its binding to the negatively charged sialic acid residues of the capsule (Oriol et al 1993). Since uterocalin has also been found in trophoblast cells (Crossett et al 1996, Ellenberger et al 2008, some of the molecule clearly passes through the capsule and presumably fulfils a role as a carrier protein. This study suggests that uterocalin also contributes to the structure of the capsule and may play a role in the initial aggregation and cross-linking of trophectoderm-produced OC-1-reactive glycoproteins.…”

Section: Development Of Horse Embryosmentioning

confidence: 99%

Influence of the uterine environment on the development of in vitro-produced equine embryos

et al. 2012

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The necessity for early interaction between the embryo and the oviductal and/or uterine environment in the horse is reflected by several striking differences between equine embryos that develop in vivo and those produced in vitro. Better understanding of the salient interactions may help to improve the efficiency of in vitro equine embryo production. In an initial experiment, cleavage-stage in vitro-produced (IVP) equine embryos were transferred into the uterus of recipient mares that had ovulated recently to determine whether premature placement in this in vivo environment would improve subsequent development. In a second experiment, an important element of the uterine environment was mimicked by adding uterocalin, a major component of the endometrial secretions during early pregnancy, to the culture medium. Intrauterine transfer of cleavage-stage IVP equine embryos yielded neither ultrasonographically detectable pregnancies nor day 7 blastocysts, indicating that the uterus is not a suitable environment for precompact morula stage horse embryos. By contrast, exposure to uterocalin during IVP improved capsule formation, although it did not measurably affect the development or expression of a panel of genes known to differ between in vivo and in vitro embryos. Further studies are required to evaluate whether uterocalin serves purely as a carrier protein or more directly promotes improved capsule development.

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Immunolocalisation of the uterine secretory proteins uterocalin, uteroferrin and uteroglobin in the mare's uterus and placenta throughout pregnancy

Cited by 43 publications

References 35 publications

Effects of long-term progesterone on developmental and functional aspects of porcine uterine epithelia and vasculature: progesterone alone does not support development of uterine glands comparable to that of pregnancy

Effects of long-term progesterone on developmental and functional aspects of porcine uterine epithelia and vasculature: progesterone alone does not support development of uterine glands comparable to that of pregnancy

Effects of long-term progesterone exposure on porcine uterine gene expression: progesterone alone does not induce secreted phosphoprotein 1 (osteopontin) in glandular epithelium

Influence of the uterine environment on the development of in vitro-produced equine embryos

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