Effect of Intraluteal Injection of Endothelin Type A Receptor Antagonist on PGF2.ALPHA.-induced Luteolysis in the Cow

Watanabe, Sho; Shirasuna, Koumei; Matsui, Motozumi; Yamamoto, Daisuke; Berisha, Bajram; Schams, D.; Miyamoto, Akio

doi:10.1262/jrd.18018

Cited by 38 publications

(33 citation statements)

References 41 publications

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“…Total RNA was extracted following the protocol of Chomczynski and Sacchi using TRIzol reagent [19] as in our previous study [20]. The extracted total RNA was stored in RNA storage solution (Ambion, Austin, TX, USA) at -80 C until use for cDNA production.…”

Section: Rna Isolation and Real-time Pcrmentioning

confidence: 99%

“…The extracted total RNA was stored in RNA storage solution (Ambion, Austin, TX, USA) at -80 C until use for cDNA production. RNA samples were treated with DNase using an RQ1 RNase-Free DNase kit (Promega, Madison, WI, USA) as in our previous study [20]. The synthesized cDNA was stored at -30 C.…”

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confidence: 99%

“…RNA samples were treated with DNase using an RQ1 RNase-Free DNase kit (Promega, Madison, WI, USA) as in our previous study [20]. The synthesized cDNA was stored at -30 C.The mRNA expression of ISG15, OAS-1, IL-10, TNFα, and β-actin was quantified by real-time PCR with a LightCycler (Roche Diagnostics GmbH, Mannheim, Germany) as described in our previous study [20]. The primers used for real-time PCR were as follows: 5′-ggtatgatgcgagctgaagcactt-3′, forward, and 5′-acctccctgctgtcaaggt-3′, reverse, for ISG15 (Accession No.…”

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Upregulation of Interferon-stimulated Genes and Interleukin-10 in Peripheral Blood Immune Cells During Early Pregnancy in Dairy Cows

et al. 2012

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Abstract. In cows, interferon-tau (IFNT) regulates maternal recognition around days 15-19 after artificial insemination (AI). The present study hypothesized that if key target genes of IFNT are clearly upregulated in earlier stages of pregnancy, these genes could be use as indices of future pregnancy in cows. Therefore, we determined the expression of these genes in peripheral blood mononuclear leukocytes (PBMCs) and polymorphonuclear granulocytes (PMNs) during the maternal recognition period (MRP). Twenty multiparous Holstein cows were subjected to AI on day 0 and categorized into the following groups: pregnancy (Preg, n = 9), embryonic death (ED, n = 5) and non-pregnancy (NP, n = 6). Progesterone levels in the Preg group were higher than those in the NP group on days 12-21. ISG15 and OAS-1 (IFN-stimulated genes: ISGs) mRNA in PBMCs on day 8 was higher in the Preg group than in the NP group, and these mRNAs in PMNs was higher in the Preg group on day 5 than in the NP and ED groups. Interleukin-10 (IL-10, Th2 cytokine) mRNA expression increased on day 8 in the PBMCs of pregnant cows. Tumor necrosis factor α (TNFα, Th1 cytokine) mRNA expression was stable in all groups. In an in vitro cell culture experiment, IFNT stimulated mRNA expression of ISGs in both PBMCs and PMNs. IFNT stimulated IL-10 mRNA expression in PBMCs, whereas IFNT increased TNFα mRNA levels in PBMCs in vitro. The results suggest that ISGs and IL-10 could be responsive to IFNT before the MRP in peripheral blood immune cells and may be useful target genes for reliable indices of pregnancy before the MRP. Key words: Cow, Immune cells, Interferon tau, Pregnancy (J. Reprod. Dev. 58: [84][85][86][87][88][89][90] 2012) D uring the past 5 decades, milk production per cow has dramatically increased because of improved management, nutrition, and genetic selection [1,2]. In contrast, the decreases in fertility and conception rates of the modern high-producing dairy cow are the major causes of economic loss for dairy producers [2,3]. After establishment of pregnancy in domestic ruminants, the conceptus secretes interferon-tau (IFNT) as a maternal recognition factor [4]. IFNT acts in the uterus around day 16 after insemination and prevents luteolysis by inhibiting prostaglandin F 2α release, resulting in the maintenance of corpus luteum function; therefore, this period is termed the maternal recognition period (MRP) [5]. IFNT induces the synthesis and secretion of IFN-stimulated genes (ISGs) such as ISG15, 2′,5′-oligoadenylate synthetase (OAS-1), IFN regulatory factor 1, Mx1 and Mx2 not only in the uterus but also in blood cells in ewes and cows [6][7][8][9][10]. In fact, in bovine peripheral blood leukocytes, ISG15 mRNA levels were higher in pregnant cows than in nonpregnant cows on days 18 and 20 after artificial insemination (AI) [7][8][9]. Technology for the early detection of pregnancy is needed to identify nonpregnant cows and to synchronize and artificially inseminate these cows prior to the next ovulation, and many studies have focused on identify...

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Section: Rna Isolation and Real-time Pcrmentioning

confidence: 99%

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Upregulation of Interferon-stimulated Genes and Interleukin-10 in Peripheral Blood Immune Cells During Early Pregnancy in Dairy Cows

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Section: Capillary Tube Formation On Matrigel Of Lyecsmentioning

confidence: 99%

“…The cDNA was synthesized as described previously (Watanabe et al 2006). The mRNA expression levels of ISG15, LYVE1, podoplanin, VEGFR3, VEGFC, VEGFD, b-actin, and GAPDH were quantified by RT quantitative PCR (RT-qPCR) as described previously (Watanabe et al 2006). RT-qPCRs were performed in duplicate in a final volume of 10 ml containing 5 ml SYBER Green, 2.8 ml of H 2 O, 0.1 ml of 50 mM forward and reverse primers (Table 1 lists primer sequences and accession numbers), and 2 ml cDNA template or water (as nontemplate negative control).…”

Section: Capillary Tube Formation On Matrigel Of Lyecsmentioning

confidence: 99%

Possible involvement of IFNT in lymphangiogenesis in the corpus luteum during the maternal recognition period in the cow

Nitta¹,

Shirasuna²,

Haneda³

et al. 2011

Reproduction

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The corpus luteum (CL), which secretes large amounts of progesterone and is thus essential for establishing pregnancy, contains various types of immune cells that may play essential roles in CL function by generating immune responses. The lymphatic system is the second circulation system and is necessary for immune function, but the lymphatic system of the bovine CL has not been characterized in detail. We collected bovine CLs on days 12 and 16 of the estrous cycle (C12 and C16) and days 16 and 40 of early pregnancy (P16 and P40). Lymphatic endothelial hyaluronan receptor 1 (LYVE1) protein was detected in the CL by immunohistochemistry and western blotting and increased at P40 compared with C16. The mRNA expression levels of lymphangiogenic factors, such as vascular endothelial growth factor-C (VEGFC), VEGFD, and their common receptor VEGFR3, as well as the lymphatic endothelial cell (LyEC) marker podoplanin, increased in P16 and P40 CLs. Thus, it is suggested that the lymphatic system of the bovine CL reconstitutes during early pregnancy. Interferon tau (IFNT) from the conceptus in the uterus is a candidate for activating luteal lymphangiogenesis during the maternal recognition period (MRP). We found that treatment of LyECs isolated from internal iliac lymphatic vessels with IFNT stimulated LyEC proliferation and significantly increased mRNA expression of VEGFC and IFN-stimulated gene 15. Moreover, both IFNT and VEGFC induced LyECs to form capillary-like tubes in vitro.In conclusion, it is suggested that new lymphangiogenesis in the bovine CL begins during the MRP and that IFNT may mediate this novel phenomenon.

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Effect of local neutralization of basic fibroblast growth factor or vascular endothelial growth factor by a specific antibody on the development of the corpus luteum in the cow

Yamashita

Kamada

Shirasuna

et al. 2008

Molecular Reproduction Devel

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Active angiogenesis and progesterone (P) synthesis occur in parallel during development of the corpus luteum (CL). Basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) are known to stimulate angiogenesis and P synthesis in vitro. The aim of the present study was to investigate the impact of bFGF or VEGF on the CL development in the cow by using a specific antibody against bFGF or VEGF. bFGF antibody, VEGF antibody, or saline as a control (n = 4 cows/treatment) were injected directly into the CL immediately after ovulation (Day 1), and the treatment was continued for 3 times/day over 7 days. Luteal biopsies were applied on Day 8 of the estrous cycle to determine the expression of genes associated with P synthesis and angiogenesis. Intraluteal injections with the bFGF antibody or the VEGF antibody markedly decreased the CL volume, plasma P concentration and StAR mRNA expression. bFGF antibody treatment decreased the mRNA expression of bFGF, FGF receptor-1, VEGF120, and angiopoietin (ANPT)-1, and increased ANPT-2/ANPT-1 ratio. However, VEGF antibody treatment decreased ANPT-2 mRNA expression and ANPT-2/ANPT-1 ratio. These results indicate that local neutralization of bFGF or VEGF changes genes regulating angiogenesis and P synthesis, and remarkably suppresses the CL size and P secretion during the development of CL in the cow, supporting the concept that bFGF and VEGF control the CL formation and function.

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Effect of Intraluteal Injection of Endothelin Type A Receptor Antagonist on PGF2.ALPHA.-induced Luteolysis in the Cow

Cited by 38 publications

References 41 publications

Upregulation of Interferon-stimulated Genes and Interleukin-10 in Peripheral Blood Immune Cells During Early Pregnancy in Dairy Cows

Upregulation of Interferon-stimulated Genes and Interleukin-10 in Peripheral Blood Immune Cells During Early Pregnancy in Dairy Cows

Possible involvement of IFNT in lymphangiogenesis in the corpus luteum during the maternal recognition period in the cow

Effect of local neutralization of basic fibroblast growth factor or vascular endothelial growth factor by a specific antibody on the development of the corpus luteum in the cow

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