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Identification of 9 uterine genes that are regulated during mouse pregnancy and exhibit abnormal levels in the cyclooxygenase-1 knockout mouse.

Zhao B, Koon D, Curtis AL, Soper J, Bethin KE - Reprod. Biol. Endocrinol. (2007)

Bottom Line: There are several putative transcription factor binding sites that are shared by many of the 9 genes identified here including; estrogen and progesterone response elements and Ets binding sites.In summary, this work identifies 9 uterine murine genes that may play a role in parturition.The function of these genes is consistent with an important role of the immune system in parturition.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pediatrics and Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Riley Hospital for Children, Indianapolis, IN 46202, USA. baohuiz@yahoo.com

ABSTRACT

Background: Preterm birth is the leading cause of all infant mortality. In 2004, 12.5% of all births were preterm. In order to understand preterm labor, we must first understand normal labor. Since many of the myometrial changes that occur during pregnancy are similar in mice and humans and mouse gestation is short, we have studied the uterine genes that change in the mouse during pregnancy. Here, we used microarray analysis to identify uterine genes in the gravid mouse that are differentially regulated in the cyclooxygenase-1 knockout mouse model of delayed parturition.

Methods: Gestational d18.0 uteri (n = 4) were collected from pregnant wild-type and cyclooxygenase-1 knockout mice. Part of the uterus was used for frozen sections and RNA was isolated from the remainder. Microarray analysis was performed at the Indiana University School of Medicine Genomic Core and analyzed using the Microarray Data Portal. Northern analysis was performed to confirm microarray data and the genes localized in the gravid uterus by in situ hybridization.

Results: We identified 277 genes that are abnormally expressed in the gravid d18.0 cyclooxygenase-1 knockout mouse. Nine of these genes are also regulated in the normal murine uterus during the last half of gestation. Many of these genes are involved in the immune response, consistent with an important role of the immune system in parturition. Expression of 4 of these genes; arginase I, IgJ, Tnfrsf9 and troponin; was confirmed by Northern analysis to be mis-regulated during pregnancy in the knockout mouse. In situ hybridization of these genes demonstrated a similar location in the gravid wild-type and Cox-1 knockout mouse uteri.

Conclusion: To our knowledge, this is the first work to demonstrate the uterine location of these 4 genes in the mouse during late pregnancy. There are several putative transcription factor binding sites that are shared by many of the 9 genes identified here including; estrogen and progesterone response elements and Ets binding sites. In summary, this work identifies 9 uterine murine genes that may play a role in parturition. The function of these genes is consistent with an important role of the immune system in parturition.

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Bar graphs for Northern analysis of arginase, immunoglobulin J chain, Tnfrsf9 and troponin. Ten μg of RNA isolated from gravid d13.5, d16.5 and d19.0 wild-type (n = 3) and d13.5, d16.5, d19.0, d20.0 and d21.0 Cox-1 KO (n = 3) uteri were run on the gel. Error bars show the standard error.
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Figure 5: Bar graphs for Northern analysis of arginase, immunoglobulin J chain, Tnfrsf9 and troponin. Ten μg of RNA isolated from gravid d13.5, d16.5 and d19.0 wild-type (n = 3) and d13.5, d16.5, d19.0, d20.0 and d21.0 Cox-1 KO (n = 3) uteri were run on the gel. Error bars show the standard error.

Mentions: From the 9 candidate genes we chose the 3 genes that were up-regulated the most and one of the genes that was down-regulated the most in the Cox-1 KO mice compared to the wild-type mice to study further by Northern analysis: arginase I (liver type), immunoglobulin J chain (IgJ), tumor necrosis factor receptor superfamily 9 (Tnfrsf9) and troponin I (Tnni2) (skeletal, fast) (Fig. 1). In situ hybridization showed that all 4 genes were expressed in the same location and distribution in the Cox-1 KO mouse and the wild-type mice (Figs. 2, 3, 4). There was no hybridization signal detected for any of the sense probes for these genes (data not shown). Northern analysis was also done on gravid uterus from wild-type d13.5, d16.5 and d19.0 (n = 3) and Cox-1 KO d13.5, d16.5, d19.0, d20.0 and d21.0 (n = 3) mice (Fig. 5). The results were consistent with the relative expression levels demonstrated on the microarray analysis for the wild-type mice. Northern analysis was not done on the remaining genes. However, in situ hybridization was performed on Klk6, Gem, Cftr and Atp8a1 (data not shown) and the signal intensity qualitatively agreed with the microarray data. In situ hybridization of arginase, Tnfrsf9 and Tnni2 d18.0 looked similar to d19.0 in situs (data not shown).


Identification of 9 uterine genes that are regulated during mouse pregnancy and exhibit abnormal levels in the cyclooxygenase-1 knockout mouse.

Zhao B, Koon D, Curtis AL, Soper J, Bethin KE - Reprod. Biol. Endocrinol. (2007)

Bar graphs for Northern analysis of arginase, immunoglobulin J chain, Tnfrsf9 and troponin. Ten μg of RNA isolated from gravid d13.5, d16.5 and d19.0 wild-type (n = 3) and d13.5, d16.5, d19.0, d20.0 and d21.0 Cox-1 KO (n = 3) uteri were run on the gel. Error bars show the standard error.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC1941732&req=5

Figure 5: Bar graphs for Northern analysis of arginase, immunoglobulin J chain, Tnfrsf9 and troponin. Ten μg of RNA isolated from gravid d13.5, d16.5 and d19.0 wild-type (n = 3) and d13.5, d16.5, d19.0, d20.0 and d21.0 Cox-1 KO (n = 3) uteri were run on the gel. Error bars show the standard error.
Mentions: From the 9 candidate genes we chose the 3 genes that were up-regulated the most and one of the genes that was down-regulated the most in the Cox-1 KO mice compared to the wild-type mice to study further by Northern analysis: arginase I (liver type), immunoglobulin J chain (IgJ), tumor necrosis factor receptor superfamily 9 (Tnfrsf9) and troponin I (Tnni2) (skeletal, fast) (Fig. 1). In situ hybridization showed that all 4 genes were expressed in the same location and distribution in the Cox-1 KO mouse and the wild-type mice (Figs. 2, 3, 4). There was no hybridization signal detected for any of the sense probes for these genes (data not shown). Northern analysis was also done on gravid uterus from wild-type d13.5, d16.5 and d19.0 (n = 3) and Cox-1 KO d13.5, d16.5, d19.0, d20.0 and d21.0 (n = 3) mice (Fig. 5). The results were consistent with the relative expression levels demonstrated on the microarray analysis for the wild-type mice. Northern analysis was not done on the remaining genes. However, in situ hybridization was performed on Klk6, Gem, Cftr and Atp8a1 (data not shown) and the signal intensity qualitatively agreed with the microarray data. In situ hybridization of arginase, Tnfrsf9 and Tnni2 d18.0 looked similar to d19.0 in situs (data not shown).

Bottom Line: There are several putative transcription factor binding sites that are shared by many of the 9 genes identified here including; estrogen and progesterone response elements and Ets binding sites.In summary, this work identifies 9 uterine murine genes that may play a role in parturition.The function of these genes is consistent with an important role of the immune system in parturition.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pediatrics and Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Riley Hospital for Children, Indianapolis, IN 46202, USA. baohuiz@yahoo.com

ABSTRACT

Background: Preterm birth is the leading cause of all infant mortality. In 2004, 12.5% of all births were preterm. In order to understand preterm labor, we must first understand normal labor. Since many of the myometrial changes that occur during pregnancy are similar in mice and humans and mouse gestation is short, we have studied the uterine genes that change in the mouse during pregnancy. Here, we used microarray analysis to identify uterine genes in the gravid mouse that are differentially regulated in the cyclooxygenase-1 knockout mouse model of delayed parturition.

Methods: Gestational d18.0 uteri (n = 4) were collected from pregnant wild-type and cyclooxygenase-1 knockout mice. Part of the uterus was used for frozen sections and RNA was isolated from the remainder. Microarray analysis was performed at the Indiana University School of Medicine Genomic Core and analyzed using the Microarray Data Portal. Northern analysis was performed to confirm microarray data and the genes localized in the gravid uterus by in situ hybridization.

Results: We identified 277 genes that are abnormally expressed in the gravid d18.0 cyclooxygenase-1 knockout mouse. Nine of these genes are also regulated in the normal murine uterus during the last half of gestation. Many of these genes are involved in the immune response, consistent with an important role of the immune system in parturition. Expression of 4 of these genes; arginase I, IgJ, Tnfrsf9 and troponin; was confirmed by Northern analysis to be mis-regulated during pregnancy in the knockout mouse. In situ hybridization of these genes demonstrated a similar location in the gravid wild-type and Cox-1 knockout mouse uteri.

Conclusion: To our knowledge, this is the first work to demonstrate the uterine location of these 4 genes in the mouse during late pregnancy. There are several putative transcription factor binding sites that are shared by many of the 9 genes identified here including; estrogen and progesterone response elements and Ets binding sites. In summary, this work identifies 9 uterine murine genes that may play a role in parturition. The function of these genes is consistent with an important role of the immune system in parturition.

Show MeSH
Related in: MedlinePlus