Expression of rat uterine serine proteinases homologous to mouse implantation serine proteinase 2

Oozono, Shinji; Yamauchi, Nobuhiko; Nishimura, Kyohei; Matsumoto, Kenji; Watanabe, Ryoji; Kubota, Kaiyu; Aramaki, Shigeto; Sato, Fuminori; Wood, Chris; Soh, Tomoki; Kizaki, Kei Ichirou; Hattori, Masa Aki

doi:10.1002/jez.b.21237

Cited by 6 publications

(4 citation statements)

References 32 publications

(22 reference statements)

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“…The UESCs were isolated from Per2-dLuc transgenic rats on day 4.5 of gestation, as reported previously (22,39,41). The uterine lumens were filled with PBS containing 0.1% collagenase and incubated at 37°C for 1 h in a shaking water bath.…”

Section: Methodsmentioning

confidence: 99%

Inhibitory role of REV-ERBα in the expression of bone morphogenetic protein gene family in rat uterus endometrium stromal cells

Tasaki

Zhao

Isayama

et al. 2015

American Journal of Physiology-Cell Physiology

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Uterus circadian rhythms have been implicated in the gestation processes of mammals through entraining of the clock proteins to numerous downstream genes. Bone morphogenetic proteins (BMPs), having clock-controlled regulatory sites in their gene promoters, are expressed in the uterus during decidualization, but the regulation of the Bmp gene expression is poorly understood. The present study was designed to dissect the physiological roles of the uterus oscillators in the Bmp expression using the uterus endometrial stromal cells (UESCs) isolated from Per2-dLuc transgenic rats on day 4.5 of gestation. The in vitro decidualization of UESCs was induced by medroxyprogesterone acetate and 2-O-dibutyryl cAMP. A significant decline of Per2-dLuc bioluminescence activity was induced in decidual cells, and concomitantly, the expression of canonical clock genes was downregulated. Conversely, the expression of the core Bmp genes Bmp2, Bmp4, Bmp6, and Bmp7 was upregulated. In UESCs transfected with Bmal1-specific siRNA, in which Rev-erbα expression was downregulated, Bmp genes, such as Bmp2, Bmp4, and Bmp6 were upregulated. However, Bmp1, Bmp7, and Bmp8a were not significantly affected by Bmal1 silencing. The expression of all Bmp genes was enhanced after treatment with the REV-ERBα antagonist (SR8278), although their rhythmic profiles were differed from each other. The binding of REV-ERBα to the proximal regions of the Bmp2 and Bmp4 promoters was revealed by chromatin immunoprecipitation-PCR analysis. Collectively, these results indicate that the Bmp genes are upregulated by the attenuation of the cellular circadian clock; in particular, its core component REV-ERBα functions as a transcriptional silencer in the Bmp gene family.

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Section: Methodsmentioning

confidence: 99%

Inhibitory role of REV-ERBα in the expression of bone morphogenetic protein gene family in rat uterus endometrium stromal cells

Tasaki

Zhao

Isayama

et al. 2015

American Journal of Physiology-Cell Physiology

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“…Preparation and Culture of the UESCs UESCs were isolated from Per2-dLuc transgenic rats on day 4.50 and day 6.50 of gestation as reported previously (He et al, 2007b;Matsumoto et al, 2009;Oozono et al, 2008). The uterine lumens were filled with phosphatebuffered saline (PBS) containing 0.1% collagenase and incubated at 37.8°C for 1 h in a shaking water bath.…”

Section: Animalsmentioning

confidence: 99%

Down-Regulation of Circadian Clock GenePeriod 2in Uterine Endometrial Stromal Cells of Pregnant Rats During Decidualization

Uchikawa

Kawamura

Yamauchi

et al. 2010

Chronobiology International

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Circadian rhythms are modulated in a variety of peripheral tissues, including in the uterus where endometrial stromal cells (UESCs) undergo proliferation and differentiation (decidualization) during gestation. Here the authors focused on circadian rhythms in UESCs during implantation and decidualization in rodents. As revealed by analyses of cultured UESCs from pregnant Per2 promoter-dLuc transgenic rats, Per2 oscillation of ∼24 h was observed in response to dexamethasone. Per2 oscillation was enhanced in UESCs during implantation, whereas they were attenuated during decidualization. In vivo studies showed that PER2 protein in the uteri displayed a peak at zeitberger time 4 (ZT 4) (day 4.50 of gestation) and a trough at ZT 12 (day 4.83), indicating its circadian rhythmicity. Conversely, no significant circadian rhythm of the PER2 protein was observed during decidualization. Fluorescent immunohistochemical studies also supported circadian rhythmicity of the PER2 protein in its intracellular distribution. In accordance with Per2 mRNA expression, a circadian rhythm of vascular endothelial growth factor (Vegf) gene expression, having several E-box or E-box-like sites at the upstream of the transcription start site, was observed during implantation, showing a peak at ZT 0 and a trough at ZT 12. In contrast, Vegf mRNA expression displayed no circadian rhythm during decidualization. Collectively, the present results prove that Per2 oscillation is down-regulated in UESCs during decidualization. It is strongly suggested that cellular differentiation in UESCs interferes with circadian clockwork.

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“…The UESCs were isolated from Per2-dLuc transgenic rats on day 4.50 of gestation as reported previously (6, 28, 29). The harvested cells were washed thrice with fresh DMEM/F12, and seeded onto 35 mm collagen-coated dishes at the density of 2 × 10 5 cells/dish with 2 mL of culture medium (phenol red-free DMEM/F12 supplemented with 10% charcoal-treated FBS and 1× PS).…”

Section: Methodsmentioning

confidence: 99%

Profiling of circadian genes expressed in the uterus endometrial stromal cells of pregnant rats as revealed by DNA microarray coupled with RNA interference

et al. 2013

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The peripheral circadian oscillator plays an essential role in synchronizing local physiology to operate in a circadian manner via regulation of the expression of clock-controlled genes. The present study aimed to evaluate the circadian rhythms of clock genes and clock-controlled genes expressed in the rat uterus endometrial stromal cells (UESCs) during the stage of implantation by a DNA microarray. Of 12,252 genes showing significantly expression, 7,235 genes displayed significant alterations. As revealed by the biological pathway analysis using the database for annotation, visualization, and integrated discovery online annotation software, genes were involved in cell cycle, glutathione metabolism, MAPK signaling pathway, fatty acid metabolism, ubiquitin mediated proteolysis, focal adhesion, and PPAR signaling pathway. The clustering of clock genes were mainly divided into four groups: the first group was Rorα, Timeless, Npas2, Bmal1, Id2, and Cry2; the second group Per1, Per2, Per3, Dec1, Tef, and Dbp; the third group Bmal2, Cry1, E4bp4, Rorβ, and Clock; the fourth group Rev-erbα. Eleven implantation-related genes and 24 placenta formation-related genes displayed significant alterations, suggesting that these genes involved in implantation and placenta formation are controlled under circadian clock. Some candidates as clock-controlled genes were evaluated by using RNA interference to Bmal1 mRNA. Down-regulation of Igf1 gene expression was observed by Bmal1 silencing, whereas the expression of Inhβa was significantly increased. During active oscillation of circadian clock, the apoptosis-related genes Fas and Caspase3 remained no significant changes, but they were significantly increased by knockdown of Bmal1 mRNA. These results indicate that clock-controlled genes are up- or down-regulated in rat UESCs during the stage of decidualization. DNA microarray analysis coupled with RNA interference will be helpful to understand the physiological roles of some oscillating genes in blastocyst implantation and placenta formation.

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Expression of rat uterine serine proteinases homologous to mouse implantation serine proteinase 2

Cited by 6 publications

References 32 publications

Inhibitory role of REV-ERBα in the expression of bone morphogenetic protein gene family in rat uterus endometrium stromal cells

Inhibitory role of REV-ERBα in the expression of bone morphogenetic protein gene family in rat uterus endometrium stromal cells

Down-Regulation of Circadian Clock GenePeriod 2in Uterine Endometrial Stromal Cells of Pregnant Rats During Decidualization

Profiling of circadian genes expressed in the uterus endometrial stromal cells of pregnant rats as revealed by DNA microarray coupled with RNA interference

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