Characterization of the concentration gradient of prostaglandin H synthase 2 mRNA throughout the pregnant baboon uterus

Wen, Wu; Smith, Gordon C. S.; Rose, James C.; Nathanielsz, Peter W.

doi:10.1677/joe.0.1820241

Cited by 10 publications

(9 citation statements)

References 17 publications

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“…Therefore, COX2 is the focal point of our present study. We have recently demonstrated that cervical glandular epithelial cells are the major cell type containing COX2 mRNA and protein in pregnant baboon and sheep (Wu et al 2004b(Wu et al , 2005a. Our data suggest that the cervix is not only the target of prostaglandins but also a site for producing prostaglandins.…”

Section: Introductionmentioning

confidence: 56%

Regulation of the prostaglandin enzymatic system by estradiol and progesterone in nonpregnant sheep cervix

Zhang

Collins²,

Chakrabarty³

et al. 2007

Reproduction

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In the present study, we examined the in vivo effects of estradiol (E 2 ) and progesterone on cyclooxygenase (COX) 2, prostaglandin F synthase (PTGFS, also known as PGFS), and membrane-associated prostaglandin E synthase 1 (mPTGES1) expression at both mRNA and protein levels using a nonpregnant ovariectomized (OVX) sheep model. Sixteen ewes were OVX shortly after ovulation. After 40 days, ewes were treated with saline (Cont, nZ5), or E 2 infused intravenously for 2 days (50 mg/day, nZ5) or intravaginal progesterone (P) sponges for 10 days (0.3 g P, nZ6). Cervical COX2, PTGFS, and mPTGES1 mRNA and protein were quantified by northern and western blot analyses respectively. In situ hybridization and/or immunocytochemistry were used to localize the cellular distribution of COX2, PTGFS, and mPTGES1 mRNAs and proteins. COX2 mRNA abundance increased significantly in the cervix after E 2 treatment (P!0.05). However, progesterone was a more potent stimulator than E 2 of COX2 mRNA and protein abundance in the cervix (P!0.01). In contrast, PTGFS and mPTGES1 mRNA and protein concentrations did not change after E 2 or progesterone treatment (PO0.05). COX2, PTGFS, and mPTGES1 mRNA and protein were only localized in cervical glandular epithelial cells. This study shows that increased cervical COX2 mRNA and protein, but not PTGFS and mPTGES1 mRNA and protein, were associated with E 2 and progesterone treatment in nonpregnant sheep. More strikingly, progesterone was a more potent stimulator of cervical COX2 expression than E 2 . The expression of COX2, PTGFS, and mPTGES1 mRNA and/or protein was confined in the cervical glandular epithelial cells of nonpregnant sheep.

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Section: Introductionmentioning

confidence: 56%

Regulation of the prostaglandin enzymatic system by estradiol and progesterone in nonpregnant sheep cervix

Zhang

Collins²,

Chakrabarty³

et al. 2007

Reproduction

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“…COX1 is constitutively expressed in most tissues, and COX2 is an inducible enzyme that responds to inflammatory cytokines and growth factors and is increased in intrauterine tissues during premature and spontaneous term labor [12,13]. We have recently demonstrated that cervical glandular epithelial cells are the major cell type containing COX2 mRNA and protein in pregnant baboon, which peaks near term [14]. Our data suggest that the cervix is not only the target of PGs but also a site for producing PGs.…”

Section: Introductionmentioning

confidence: 70%

“…Samples of polyadenylated RNA (2 g) were separated by electrophoresis on a 1.4% (wt/vol) agarose-0.66 M formaldehyde gel and transferred onto a nylon membrane (NEN Life Science) and then subjected to Northern blot analysis for COX2, PTGER2, PTGER4, and ESR1 mRNAs as described previously [13,14,20,22]. Cyclophilin mRNA was used to control RNA loading.…”

Section: Northern Blot Analysismentioning

confidence: 99%

“…Specificity of immunostaining for the COX2 was confirmed by two approaches: 1) omission of the primary antibody and 2) incubation of the slides with the normal rabbit serum instead of the primary antibody [13,14,20,23].…”

Section: Immunocytochemistrymentioning

confidence: 99%

“…A rabbit polyclonal antibody for human COX2 (Oxford Biomedical Research, Inc., Oxford, MI) was used at 1:1000 dilutions and incubated at 4ЊC for 20 h. This COX2 antibody has been characterized in our previous studies in application of sheep tissues [13,14,20,23]. Rabbit polyclonal antibodies for human PTGER2 and PTGER4 receptors (Cayman Chemical Co.) were used at 1:500 and 1:1000 dilutions and incubated at 4ЊC for 20 h. Specificity of Western blot analysis for PTGER2 and PTGER4 antibodies was confirmed by preabsorpting PTGER2 and PTGER4 antibodies with synthetic polypeptides that were used for generating the antibodies (Cayman Chemical).…”

Section: Antibodies For Western Blot Analysis and Immunocytochemistrymentioning

confidence: 99%

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Sufficient Progesterone-Priming Prior to Estradiol Stimulation Is Required for Optimal Induction of the Cervical Prostaglandin System in Pregnant Sheep at 0.7 Gestations1

Wen¹,

Coksaygan²,

Chakrabarty³

et al. 2005

Biology of Reproduction

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The purposes of this study were to determine the separate and interactive functions of progesterone and estradiol in regulating the cervical prostaglandin (PG) system in pregnant sheep at 0.7 gestations. At 106-108 days of gestational age (dGA), ewes were treated with vehicle for 14 days (n = 5) or vehicle for 12 days followed by estradiol 5 mg twice a day, intramuscularly for 2 days (n = 5) or progesterone 100 mg, twice a day, intramuscularly for 14 days (n = 5) or progesterone 100 mg twice a day, intramuscularly for 10 days and then 2 days vehicle followed by estradiol 5 mg twice a day intramuscularly for 2 days (n = 5). At 121-123 dGA, cervical tissues were obtained under halothane anesthesia. Cervical RNA and protein were extracted and analyzed for prostaglandin-endoperoxide synthase 2 (COX2), two PGE(2) receptors, PTGER2 and PTGER4, and estrogen receptor alpha (ESR1) by Northern and Western blot analysis. Immunocytochemistry and in situ hybridization were applied to localize cellular distribution of COX2, PTGER2, and PTGER4 in the cervix. Data were analyzed by ANOVA. COX2 and PTGER4 mRNAs and proteins were increased (P < 0.05) in ewes treated with combined estradiol and progesterone but not in ewes treated with estradiol or progesterone alone compared with controls. ESR1 mRNA was increased in ewes treated with progesterone and estradiol plus progesterone. In contrast, PTGER2 mRNA and protein remained the same after all treatments. COX2 mRNA and protein were localized only in cervical glandular epithelial cells, whereas PTGER2 and PTGER4 were localized in both cervical glandular epithelial and smooth muscle cells. In conclusion, these data suggest that additional progesterone priming at 0.7 gestations synergizes with estradiol to induce cervical COX2, PTGER4, and ESR1 and support our hypothesis that stimulation of the cervical PG system by estradiol is optimized by sufficient progesterone priming in the pregnant sheep cervix.

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A molecular signature of an arrest of descent in human parturition

Mittal

Romero

Tarca

et al. 2011

American Journal of Obstetrics and Gynecology

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Objective-This study was undertaken to identify the molecular basis of an arrest of descent.Study Design-Human myometrium was obtained from women in term labor (TL; n=29) and arrest of descent (AODes, n=21). Gene expression was characterized using Illumina® HumanHT-12 microarrays. A moderated t-test and false discovery rate adjustment were applied for analysis. Confirmatory qRT-PCR and immunoblot was performed in an independent sample set.Results-400 genes were differentially expressed between women with an AODes compared to those with TL. Gene Ontology analysis indicated enrichment of biological processes and molecular functions related to inflammation and muscle function. Impacted pathways included inflammation and the actin cytoskeleton. Overexpression of HIF1A, IL-6, and PTGS2 in AODES was confirmed. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. Conclusion-We have identified a stereotypic pattern of gene expression in the myometrium of women with an arrest of descent. This represents the first study examining the molecular basis of an arrest of descent using a genome-wide approach.

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Characterization of the concentration gradient of prostaglandin H synthase 2 mRNA throughout the pregnant baboon uterus

Cited by 10 publications

References 17 publications

Regulation of the prostaglandin enzymatic system by estradiol and progesterone in nonpregnant sheep cervix

Regulation of the prostaglandin enzymatic system by estradiol and progesterone in nonpregnant sheep cervix

Sufficient Progesterone-Priming Prior to Estradiol Stimulation Is Required for Optimal Induction of the Cervical Prostaglandin System in Pregnant Sheep at 0.7 Gestations1

A molecular signature of an arrest of descent in human parturition

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