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Tianbing Ding
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Haengseok Song Division of Neonatology, Laboratory of Reproductive Biology and Infertility, Meharry Medical College, Department of Pediatrics, Vanderbilt University Medical Center, 1125 MRB IV/Light Hall, 2215 B. Garland Avenue, Nashville, Tennessee 37232-0656, USA

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Xiaohong Wang
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Atanu Khatua Division of Neonatology, Laboratory of Reproductive Biology and Infertility, Meharry Medical College, Department of Pediatrics, Vanderbilt University Medical Center, 1125 MRB IV/Light Hall, 2215 B. Garland Avenue, Nashville, Tennessee 37232-0656, USA

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Bibhash C Paria
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Blastocyst implantation occurs in the progesterone-primed uterus of hamsters, but not in mice where the progesterone-primed uterus requires estrogen influence. Leukemia inhibitory factor (Lif), an estrogen-regulated gene in mice, is an absolutely needed cytokine for uterine receptivity and implantation in this species. This study aimed to evaluate the importance of Lif ligand-receptor signaling during uterine receptivity and implantation in hamsters. We investigated whether or not the uterine expression patterns of Lif and its receptors, Lif-r and gp130, during the periimplantation period of pregnancy and its hormonal regulation in the ovariectomized hamster correlate with some of the vital phases of uterine changes during early pregnancy. Uterine Lif, Lif-r, and gp130 mRNA expressions were examined by Northern and in situ hybridization. During the uterine preparatory phase for implantation, Lif, Lif-r, and gp130 were expressed either in the gland, luminal epithelium or both. As the implantation process began, Lif expression was minimal, but Lif-r and gp130 extended to the decidual areas. This decidual expression of Lif-r and gp130 was not dependent on the presence of the embryo since these genes were expressed in the suture-induced deciduomata. We also observed that, while the uterine Lif was induced by estrogen, Lif-r and gp130 were induced by progesterone in ovariectomized hamsters. Additionally, we show that a Lif antibody when instilled intraluminally on day 3 of pregnancy reduced the number of implantation sites. Taken together, these data suggest that Lif signaling is important for uterine receptivity and implantation in hamsters.

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Wei Lei Division of Neonatology, Division of Cardiovascular Medicine, Department of Obstetrics and Gynecology, Department of Pediatrics

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Jennifer Herington Division of Neonatology, Division of Cardiovascular Medicine, Department of Obstetrics and Gynecology, Department of Pediatrics

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Cristi L Galindo Division of Neonatology, Division of Cardiovascular Medicine, Department of Obstetrics and Gynecology, Department of Pediatrics

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Tianbing Ding Division of Neonatology, Division of Cardiovascular Medicine, Department of Obstetrics and Gynecology, Department of Pediatrics

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Naoko Brown Division of Neonatology, Division of Cardiovascular Medicine, Department of Obstetrics and Gynecology, Department of Pediatrics

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Jeff Reese Division of Neonatology, Division of Cardiovascular Medicine, Department of Obstetrics and Gynecology, Department of Pediatrics

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Bibhash C Paria Division of Neonatology, Division of Cardiovascular Medicine, Department of Obstetrics and Gynecology, Department of Pediatrics

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The mouse model has greatly contributed to understanding molecular mechanisms involved in the regulation of progesterone (P4) plus estrogen (E)-dependent blastocyst implantation process. However, little is known about contributory molecular mechanisms of the P4-only-dependent blastocyst implantation process that occurs in species such as hamsters, guineapigs, rabbits, pigs, rhesus monkeys, and perhaps humans. We used the hamster as a model of P4-only-dependent blastocyst implantation and carried out cross-species microarray (CSM) analyses to reveal differentially expressed genes at the blastocyst implantation site (BIS), in order to advance the understanding of molecular mechanisms of implantation. Upregulation of 112 genes and downregulation of 77 genes at the BIS were identified using a mouse microarray platform, while use of the human microarray revealed 62 up- and 38 down-regulated genes at the BIS. Excitingly, a sizable number of genes (30 up- and 11 down-regulated genes) were identified as a shared pool by both CSMs. Real-time RT-PCR and in situ hybridization validated the expression patterns of several up- and down-regulated genes identified by both CSMs at the hamster and mouse BIS to demonstrate the merit of CSM findings across species, in addition to revealing genes specific to hamsters. Functional annotation analysis found that genes involved in the spliceosome, proteasome, and ubiquination pathways are enriched at the hamster BIS, while genes associated with tight junction, SAPK/JNK signaling, and PPARα/RXRα signalings are repressed at the BIS. Overall, this study provides a pool of genes and evidence of their participation in up- and down-regulated cellular functions/pathways at the hamster BIS.

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Wei Lei
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Heidi Nguyen
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Naoko Brown
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Hua Ni
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Tina Kiffer-Moreira Division of Neonatology, Sanford-Burnham Medical Research Institute, Department of Pediatrics, Vanderbilt University Medical Center, 1125 Light Hall, 2215 B. Garland Avenue, Nashville, Tennessee 37232-0656, USA and

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Jeff Reese
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José Luis Millán Division of Neonatology, Sanford-Burnham Medical Research Institute, Department of Pediatrics, Vanderbilt University Medical Center, 1125 Light Hall, 2215 B. Garland Avenue, Nashville, Tennessee 37232-0656, USA and

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Bibhash C Paria
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Alkaline phosphatase (AP) activity has been demonstrated in the uterus of several species, but its importance in the uterus, in general and during pregnancy, is yet to be revealed. In this study, we focused on identifying AP isozyme types and their hormonal regulation, cell type, and event-specific expression and possible functions in the hamster uterus during the cycle and early pregnancy. Our RT-PCR and in situ hybridization studies demonstrated that among the known Akp2, Akp3, Akp5, and Akp6 murine AP isozyme genes, hamster uteri express only Akp2 and Akp6; both genes are co-expressed in luminal epithelial cells. Studies in cyclic and ovariectomized hamsters established that while progesterone (P4) is the major uterine Akp2 inducer, both P4 and estrogen are strong Akp6 regulators. Studies in preimplantation uteri showed induction of both genes and the activity of their encoded isozymes in luminal epithelial cells during uterine receptivity. However, at the beginning of implantation, Akp2 showed reduced expression in luminal epithelial cells surrounding the implanted embryo. By contrast, expression of Akp6 and its isozyme was maintained in luminal epithelial cells adjacent to, but not away from, the implanted embryo. Following implantation, stromal transformation to decidua was associated with induced expressions of only Akp2 and its isozyme. We next demonstrated that uterine APs dephosphorylate and detoxify endotoxin lipopolysaccharide at their sites of production and activity. Taken together, our findings suggest that uterine APs contribute to uterine receptivity, implantation, and decidualization in addition to their role in protection of the uterus and pregnancy against bacterial infection.

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Michael F Robuck Division of Neonatology, Department of Pediatrics, Vanderbilt University Medical Center, Nashville, Tennessee, USA

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Christine M O’Brien Department of Biomedical Engineering, Vanderbilt University, Nashville, Tennessee, USA

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Kelsi M Knapp Division of Neonatology, Department of Pediatrics, Vanderbilt University Medical Center, Nashville, Tennessee, USA

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Sheila D Shay Division of Allergy, Pulmonary, and Critical Care Medicine, Department of Medicine, Vanderbilt University Medical Center, Nashville, Tennessee, USA

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James D West Division of Allergy, Pulmonary, and Critical Care Medicine, Department of Medicine, Vanderbilt University Medical Center, Nashville, Tennessee, USA

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J M Newton Division of Maternal Fetal Medicine, Department of Obstetrics and Gynecology, Vanderbilt University Medical Center, Nashville, Tennessee, USA

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James C Slaughter Department of Biostatistics, Vanderbilt University Medical Center, Nashville, Tennessee, USA

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Bibhash C Paria Division of Neonatology, Department of Pediatrics, Vanderbilt University Medical Center, Nashville, Tennessee, USA

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Jeff Reese Division of Neonatology, Department of Pediatrics, Vanderbilt University Medical Center, Nashville, Tennessee, USA
Department of Biomedical Engineering, Vanderbilt University, Nashville, Tennessee, USA

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Jennifer L Herington Division of Neonatology, Department of Pediatrics, Vanderbilt University Medical Center, Nashville, Tennessee, USA
Department of Pharmacology, Vanderbilt University, Nashville, Tennessee, USA

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In mouse models used to study parturition or pre-clinical therapeutic testing, measurement of uterine contractions is limited to either ex vivo isometric tension or operative intrauterine pressure (IUP). The goal of this study was to: (1) develop a method for transcervical insertion of a pressure catheter to measure in vivo intrauterine contractile pressure during mouse pregnancy, (2) determine whether this method can be utilized numerous times in a single mouse pregnancy without affecting the timing of delivery or fetal outcome and (3) compare the in vivo contractile activity between mouse models of term and preterm labor (PTL). Visualization of the cervix allowed intrauterine pressure catheter (IUPC) placement into anesthetized pregnant mice (plug = day 1, delivery = day 19.5). The amplitude, frequency, duration and area under the curve (AUC) of IUP was lowest on days 16–18, increased significantly (P < 0.05) on the morning of day 19 and reached maximal levels during by the afternoon of day 19 and into the intrapartum period. An AUC threshold of 2.77 mmHg discriminated between inactive labor (day 19 am) and active labor (day 19 pm and intrapartum period). Mice examined on a single vs every experimental timepoint did not have significantly different IUP, timing of delivery, offspring number or fetal/neonatal weight. The IUP was significantly greater in LPS-treated and RU486-treated mouse models of PTL compared to time-matched vehicle control mice. Intrapartum IUP was not significantly different between term and preterm mice. We conclude that utilization of a transcervical IUPC allows sensitive assessment of in vivo uterine contractile activity and labor progression in mouse models without the need for operative approaches.

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