共查询到20条相似文献，搜索用时 93 毫秒
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Generation of recombinant bovine interferon tau in the human embryonic kidney cell line and its biological activity 
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下载免费PDF全文 Toru Takahashi Ryosuke Sakumoto Ken‐Go Hayashi Misa Hosoe Junsuke Shirai Kazuyoshi Hashizume 《Animal Science Journal》2017,88(10):1498-1505
The objective of this study was to generate recombinant bovine interferon tau (rbIFNT) in mammalian hosts. The complementary DNA encoding bovine IFNT2 was cloned for the construction of pRcRSV‐bIFNT2 expression vector. The expression vector was transfected to 293 cells. Transfected cells harboring expression vector were selected with G418. Highly expressing clonal line was adapted to serum‐free suspension culture in a spinner flask. The recombinant protein had 24 kDa apparent molecular mass, suggesting being expressed as a glycoprotein, and was purified from serum‐free conditioned medium by the combination of Diethylaminoethanol Sepharose ion exchange and Sephacryl S‐200 HR gel filtration. A total of 7.3 mg rbIFNT was obtained from 13.5 L conditioned medium. Generated rbIFNT was biologically active in terms of antiviral activity measured by the plaque inhibition assay with Madin‐Darby bovine kidney cells and the vesicular stomatitis virus. The recombinant protein was also utilized for immunization to raise antibodies in the rabbit. The generated antibody was capable of use in both Western blotting and the binding assay. The results in the present study suggest that a certain amount of rbIFNT is raised in mammalian hosts by using conventional plasmid vector and its antibody provides useful tools for studies in the biology of bovine IFNT. 相似文献
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Fuller W Bazer 《畜牧与生物技术杂志(英文版)》2013,4(1):23
Maternal recognition of pregnancy refers to the requirement for the conceptus (embryo and its associated extra-embryonic membranes) to produce a hormone that acts on the uterus and/or corpus luteum (CL) to ensure maintenance of a functional CL for production of progesterone; the hormone required for pregnancy in most mammals. The pregnancy recognition signal in primates is chorionic gonadotrophin which acts directly on the CL via luteinizing hormone receptors to ensure maintenance of functional CL during pregnancy. In ruminants, interferon tau (IFNT) is the pregnancy recognition signal. IFNT is secreted during the peri-implantation period of pregnancy and acts on uterine epithelia to silence expression of estrogen receptor alpha and oxytocin receptor which abrogates the oxytocin-dependent release of luteolytic pulses of prostaglandin F2-alpha (PGF) by uterine epithelia; therefore, the CL continues to produce progesterone required for pregnancy. Pig conceptuses secrete interferon delta and interferon gamma during the peri-implantation period of pregnancy, but there is no evidence that they are involved in pregnancy recognition signaling. Rather, pig conceptuses secrete abundant amounts of estrogens between Days 11 to 15 of pregnancy required for maternal recognition of pregnancy. Estrogen, likely in concert with prolactin, prevents secretion of PGF into the uterine venous drainage (endocrine secretion), but maintains secretion of PGF into the uterine lumen (exocrine secretion) where it is metabolized to a form that is not luteolytic. Since PGF is sequestered within the uterine lumen and unavailable to induce luteolysis, functional CL are maintained for production of progesterone. In addition to effects of chorionic gonadotrophin, IFNT and estrogens to signal pregnancy recognition, these hormones act on uterine epithelia to enhance expression of genes critical for growth and development of the conceptus. 相似文献
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Pregnancy‐Induced ISG‐15 and MX‐1 Gene Expression is Detected in the Liver of Holstein–Friesian Heifers During Late Peri‐Implantation Period 下载免费PDF全文
下载免费PDF全文 MM Meyerholz K Mense H Knaack O Sandra M Schmicke 《Reproduction in domestic animals》2016,51(1):175-177
The bovine embryonic signal interferon‐τ (IFN‐τ) produced by the trophoblast is known to pass through the uterine fluid towards the endometrium and further into the maternal blood, where IFN‐τ induces specific expression of interferon‐stimulated gene expression (ISG), for example in peripheral leucocytes. In sheep, it was shown experimentally by administration of IFN‐τ that ISG is also detectable in the liver. The objective was to test whether ISG can be detected in liver biopsy specimens from Holstein–Friesian heifers during early pregnancy. Liver biopsies were taken on day 18 from pregnant and non‐pregnant heifers (n = 19), and the interferon‐stimulated protein 15 kDa (ISG‐15) and myxovirus‐resistance protein‐1 (MX‐1) gene expression was detected. The expression of both MX‐1 (p: 24.33 ± 7.40 vs np: 9.00 ± 4.02) and ISG‐15 (p: 43.73 ± 23.22 vs 7.83 ± 3.63) was higher in pregnant compared to non‐pregnant heifers (p < 0.05). In conclusion, pregnancy induced ISG‐15 and MX‐1 gene expression in the liver already at day 18 in cattle. 相似文献
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Expression of Heparin‐Binding EGF‐Like Growth Factor (HB‐EGF) in Bovine Endometrium: Effects of HB‐EGF and Interferon‐τ on Prostaglandin Production 下载免费PDF全文
下载免费PDF全文 Heparin‐binding EGF‐like growth factor (HB‐EGF) regulates several cell functions by binding to its membrane receptor (ErbB1 and ErbB4). Experimental evidences suggest that HB‐EGF, prostaglandins (PGs) and interferon‐τ (IFN‐τ) regulate uterine function for pregnancy establishment in ruminants. In this study, the mRNA expressions of HB‐EGF, ErbB1 and ErbB4 in bovine endometrium and the effects of HB‐EGF and IFN‐τ on PGE2 and PGF2‐α production by endometrial cells were investigated. RT‐PCR analysis revealed that HB‐EGF mRNA was greater at the mid‐luteal stage than at the early and regressed luteal stages (p < 0.05). ErbB1 mRNA expression was greater at the mid‐ and late luteal stages than at the other luteal stages (p < 0.05). IFN‐τ increased the expression of HB‐EGF, ErbB1 and ErbB4 mRNA in epithelial cells (p < 0.05). HB‐EGF did not affect PGF2‐α or PGE2 production by bovine endometrial epithelial cells, but increased PGF2‐α and PGE2 production by bovine endometrial stromal cells (p < 0.05). IFN‐τ significantly decreased HB‐EGF‐stimulated PGF2‐α (p < 0.05), but not PGE2 (p > 0.05) production by stromal cells. These results indicate that HB‐EGF and its receptors expression changed in bovine endometrium throughout the oestrous cycle. IFN‐τ increased their expression in cultured endometrial cells. HB‐EGF and IFN‐τ have the ability to regulate PGs production by stromal cells and therefore may play a role in the local regulation of uterine function at the time of implantation in cattle. 相似文献
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During a 5-year period 65 Pony and 20 Thoroughbred pregnant mares were subjected to videoendoscopic hysteroscopy from 10 to 266 days of gestation. The aims of these examinations were to 1) observe foetal and placental development in vivo (60 Pony and 10 Thoroughbred mares); 2) eliminate one of unicornuate twin conceptuses (9 Thoroughbred mares); 3) recover embryonic foetal and placental tissues non-surgically for experimental purposes (47 Pony mares); 4) induce focal separation of the placenta in late gestation as an experimental model of placentitis (5 pony mares and 1 Thoroughbred mare). It was possible to view the embryo and all constituent extra-embryonic membranes of the conceptus between Days 10 and 87 of gestation without having to perforate the allantochorion. This allowed study of physiological processes such as the coordinated uterine contractions responsible for conceptus motility between Days 7 and 17, active foetal movements, which began as early as Day 34, and invasion of chorionic girdle cells into the endometrium, which occurred between Days 34 and 38. From Day 90, vision of the foetus was reduced or prevented by the increased thickness of the allantochorion. Transendoscopic recovery of the conceptus was successful in all of 10 mares under 30 days of gestation, whereas only 10 of 18 attempts between Days 30 and 45 produced the conceptus without resorting to uterine lavage after initial rupture of the allantochorion. All 9 attempts to eliminate one of unilateral twin conceptuses were unsuccessful and the technique was abandoned as clinically unsuitable. Nevertheless, 9 of 22 (41%) single conceptuses remained viable after one or more hysteroscopic examinations.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
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In the pig as in ruminant species, the implantation of the elongated conceptus - the embryo with its associated membranes - onto the maternal uterus is accompanied by an intense secretion of interferon (IFN), which culminates at day 15 of development. It has been shown that in fact the pig trophectoderm - the polarized epithelium which lines the conceptus - simultaneously secretes two types of interferons: IFN-gamma (IFN-gamma), which is the more abundant species, is produced in very substantial amounts. Another IFN is also secreted, which happens to be a novel type I IFN, now named IFN-delta. It was previously shown that the uterus is the most probable target of the pig trophoblastic IFNs, since no autocrine effect was found on the trophoblast. It has also been shown that, unlike for the ruminant species, the pig trophoblastic IFNs do not play an apparent role in the so-called maternal recognition of pregnancy. We have focused this review on IFN-gamma, because first, it is the major species secreted and secondly, IFN-gamma has various regulatory effects on different tissues, including lymphoid cells. We particularly address the question of the possible role of trophoblastic IFN-gamma in early pregnancy, in the light of the known biological functions of human and mouse IFN-gamma. 相似文献
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B Serrano‐Pérez I Garcia‐Ispierto NM de Sousa JF Beckers S Almería F López‐Gatius 《Reproduction in domestic animals》2014,49(2):275-280
Gamma interferon (IFN‐γ) production and cross‐breed pregnancy have been attributed a role in protecting dairy cows infected with Neospora caninum against abortion. Plasma levels of pregnancy‐associated glycoproteins‐1 (PAG‐1) are a marker of placental/foetal well‐being and of PAG‐2 is an abortion risk indicator in chronically N. caninum‐infected animals. The present study examines, in cross‐breed pregnancies, interactions between IFN‐γ production and levels of PAG‐1 and PAG‐2 in non‐aborting naturally Neospora‐infected dairy cows. Data were obtained from 60 pregnant Holstein‐Friesian cows: 44 Neospora‐seropositive and 16 Neospora‐seronegative; 12 became pregnant using Holstein‐Friesian semen and 48 using Limousin semen. Blood samples were collected on Days 40, 90, 120, 150, 180 and 210 of gestation. Gamma interferon was only detected in the plasma of nine of the 44 Neospora‐seropositive cows, all of them became pregnant using Limousin semen. Through GLM procedures, in cows inseminated with Limousin semen and Neospora‐seropositive cows showing no IFN‐γ production, PAG‐1 concentrations were high and increased throughout gestation compared to the levels detected in cows inseminated with Holstein‐Friesian semen and Neospora‐seropositive cows producing IFN‐γ, respectively. In Neospora‐seronegative cows and in Neospora‐seropositive cows showing no IFN‐γ production, significantly increased PAG‐2 concentrations were observed on gestation Day 120. Our findings indicate that IFN‐γ production correlates negatively and the production of antibodies against N. caninum is uncorrelated with plasma PAG concentrations during gestation in Neospora‐infected dairy cows. Accordingly, IFN‐γ production could be linked to the transplacental migration of tachyzoites, which may cause a reduction in PAG levels. 相似文献
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Agnieszka BLITEK Ewa MORAWSKA-PUCINSKA Magdalena SZYMANSKA Jolanta KIEWISZ Agnieszka WACLAWIK 《The Journal of reproduction and development》2013,59(6):512-519
Transforming growth factor (TGF) β and its receptors are expressed at the
conceptus-maternal interface during early pregnancy in the pig. The present studies
were conducted to examine: (1) the effect of conceptus products on TGFβ1 mRNA
expression and protein concentration in the porcine endometrium using in
vivo and in vitro models, and (2) the effect of TGFβ1 on
proliferation of porcine trophoblast cells in vitro. During
in vivo experiments, gilts with one surgically detached uterine
horn were slaughtered on days 11 or 14 of the estrous cycle and pregnancy. For
in vitro studies, endometrial explants and luminal epithelial
(LE) cells co-cultured with stromal (ST) cells were treated with conceptus-exposed
medium (CEM). Moreover, porcine trophoblast cells were treated with TGFβ1, and the
number of viable cells was measured. On day 11, the presence of conceptuses had no
effect on TGFβ1 mRNA expression, but decreased the TGFβ1 protein concentration in the
connected uterine horn compared with the detached uterine horn. In contrast to day
11, on day 14 after estrus, TGFβ1 mRNA expression and protein content in the
endometrium collected from the gravid uterine horn were greater when compared with
the contralateral uterine horn. The treatment of endometrial slices with CEM resulted
in greater TGFβ1 mRNA expression and protein secretion. LE cells responded to CEM
with an increased TGFβ1 mRNA level. Moreover, TGFβ1 stimulated the proliferation of
day 14 trophoblast cells. In summary, porcine conceptuses may regulate TGFβ1
synthesis in the endometrium at the time of implantation. TGFβ1, in turn, may promote
conceptus development by increasing the proliferation of trophoblast cells. 相似文献
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Zhi Chen Xin Xu Taoling Tan Daijie Chen Huanjie Liang Kaidi Sun Mingxun Li Huimin Zhang Yongjiang Mao Zhangping Yang 《Reproduction in domestic animals》2019,54(6):882-891
Dairy cow mastitis is a detrimental factor in milk quality and food safety. Mastitis generally refers to inflammation caused by infection by pathogenic microorganisms. Our studies in recent years have revealed the role of miRNA regulation in Staphylococcus aureus‐induced mastitis. In the present study, we overexpressed and suppressed miR‐145 to investigate the function of miR‐145 in Mac‐T cells. Flow cytometry, ELISA and EdU staining were used to detect changes in the secretion of several Mac‐T cytokines and in cell proliferation. We found that overexpression of miR‐145 in Mac‐T cells significantly reduced the secretion of IL‐12 and TNF‐α, but increased the secretion of IFN‐γ; the proliferation of bovine mammary epithelial cells was also inhibited. Using quantitative real‐time PCR (qRT‐PCR), Western blotting and luciferase multiplex verification techniques, we found that miR‐145 targeted and regulated FSCN1. Knock‐down of FSCN1 significantly increased the secretion of IL‐12, while the secretion of TNF‐α was significantly downregulated in Mac‐T cells. Upon S. aureus infection of mammary gland tissue, the body initiated inflammatory responses; Bta‐miR‐145 expression was downregulated, which reduced the inhibitory effect on the FSCN1 gene; and upregulation of FSCN1 expression promoted mammary epithelial cell proliferation to allow the recovery of damaged tissue. The results of the present study will aid in understanding the immune mechanism opposing S. aureus infection in dairy cows and will provide a laboratory research basis for the prevention and treatment of mastitis. 相似文献
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Hickman CF Ainslie A Ealy AD Ashworth CJ Rooke JA 《Reproduction in domestic animals》2011,46(4):608-615
This experiment examined the effects of including recombinant ovine granulocyte-macrophage colony-stimulating factor (GMCSF) in in vitro culture on secretion of interferon-τ (IFNT) by bovine blastocysts. At 32 h post-insemination (p.i.), cleaved bovine zygotes were selected and incubated with or without GMCSF for either 48 h only (between 32 and 80 h p.i., Early) or until day 9 p.i. (Throughout). Concentrations of GMCSF (ng/ml) examined were as follows: Experiment 1: 2, 5, 10 and 50 (Early only); Experiment 2: 50 (Early and Throughout); Experiment 3: 2 and 10 (Early and Throughout). In none of the experiments did GMCSF have an effect (p > 0.05) on the numbers of blastocysts formed or blastocyst characteristics as assessed by cell number, proportion of apoptotic cells or oxidation of pyruvate. When GMCSF was included in culture medium between 32 and 80 h p.i. (Early), IFNT concentrations were lower (in media drops recovered after culture of groups of embryos for 48 h between days 7 and 9 p.i. and normalized by the numbers of blastocysts developing within each drop) compared to no inclusion of GMCSF or GMCSF present Throughout culture (Experiment 2, p > 0.05; Experiment 3, p = 0.038). IFNT was present in media drops in which groups of embryos had been incubated between days 7 and 9 p.i. but in which no blastocysts had developed. Experimental treatment did not influence (p > 0.05) IFNT secretion by blastocysts incubated individually for 24 h. However, during the 24-h individual culture, blastocysts recovered on day 7 secreted less IFNT than blastocysts recovered on day 8 (mean ± SE; 15 ± 1.3 v 30 ± 3.6 pg/ml; p < 0.001). In conclusion, in contrast to previous studies in the ovine, GMCSF did not increase IFNT secretion but in agreement with the ovine did not affect bovine blastocyst development. 相似文献