118. THE PRORENIN RECEPTOR/PLZF PATHWAY IN HUMAN AMNION

2010 ◽  
Vol 22 (9) ◽  
pp. 36
Author(s):  
K. G. Pringle ◽  
A. L. Conquest ◽  
C. M. Mitchell ◽  
T. Zakar ◽  
E. R. Lumbers
Keyword(s):  
New Zealand ◽  
Amniotic Fluid ◽  
Mrna Levels ◽  
The Novel ◽  
Quantitative Real Time Pcr ◽  
Human Amnion ◽  
Reproductive Tissues ◽  
Prorenin Receptor ◽  
Human Reproductive ◽  
Neonatal Physiology

Prorenin, despite being inactive, is the major form of renin found in amniotic fluid and reproductive tissues. Prorenin becomes active if it binds to the novel prorenin receptor (ATP6AP2). The prorenin-ATP6AP2 complex has been found to stimulate translocation of Promyelocytic Zinc Finger (PLZF) protein to the nucleus where it increases expression of the p85α subunit of PI3 kinase (PI3K-p85α) and represses the expression of ATP6AP21. Progesterone and glucocorticoids have also been shown to stimulate PLZF2, 3. We aimed to find out if PLZF and the prorenin-ATP6AP2 pathway interact in human reproductive tissues. Human amnion was cultured for 24 h in media containing vehicle, dexamethasone, amniotic fluid or recombinant human (rh) prorenin. Total RNA was extracted using TRIZol® and converted to cDNA for quantitative real-time PCR using SuperScript III and random hexamers. mRNA abundances for PLZF, PI3K-p85α and ATP6AP2 were calculated relative to Alien RNA using the ΔΔCT method. Our preliminary data show that exposure of amnion explants to dexamethasone upregulates PLZF and PI3K-p85α mRNA but has no effect on ATP6AP2. Culture of amnion explants with amniotic fluid also increases PLZF but does not change PI3K or ATP6AP2. In contrast, culture of amnion explants with (rh) prorenin increases PI3K mRNA but not PLZF or ATP6AP2. As expected, dexamethasone affects PLZF expression, however in amnion there is no interaction with the ATP6AP2 pathway. In addition, we believe we have identified a novel prorenin/ATP6AP2 signalling pathway which acts on PI3K-p85α independent of PLZF. In contrast to these data, amniotic fluid increases PLZF but not PI3K-p85α mRNA levels suggesting that amniotic fluid contains other factors that oppose prorenin and glucocorticoid effects on PI3K-p85α. (1) Schefe, J.H., et al. Circ Res, 2006. 99(12): 1355–1366.(2) Fahnenstich, J., et al. Mol. Hum. Reprod., 2003. 9(10): 611–623.(3) Conquest, A. et al. Fetal and Neonatal Physiology Workshop, 2010. Wellington, New Zealand.

311. REGULATION OF THE RENIN ANGIOTENSIN SYSTEM (RAS) IN A TROPHOBLAST CELL LINE BY CYCLIC ADENOSINE MONOPHOSPHATE (cAMP) AND 5'-AZA-2'-DEOXYCYTIDINE (AZA)

2010 ◽  
Vol 22 (9) ◽  
pp. 111
Author(s):  
Y. Wang ◽  
K. G. Pringle ◽  
Y. Chen ◽  
T. Zakar ◽  
E. R. Lumbers
Keyword(s):  
Dna Methylation ◽  
New Zealand ◽  
Cell Line ◽  
Messenger Rna ◽  
Trophoblast Cell ◽  
Mrna Levels ◽  
Human Trophoblast ◽  
Downstream Target ◽  
Prorenin Receptor ◽  
Trophoblast Cell Line

Renin and renin-like activities have been detected in early gestation placentae1,2. To explore what regulates RAS expression in trophoblasts we studied the effects of cAMP and AZA (DNA demethylating agent) on the expression of mRNAs for prorenin receptor (ATP6AP2), prorenin (REN), angiotensinogen (AGT), angiotensin converting enzyme (ACE), ACE2, angiotensin II type 1 receptor (AGTR1), and a downstream target of the prorenin receptor, cyclooxygenase-2 (PTGS2) in the early human trophoblast cell line HTR-8/SVneo. Cells were cultured for 24 or 48 h with vehicle, 0.5 mM cAMP or 15 mM AZA. Messenger RNA abundances were measured relative to Alien RNA using real-time qRT-PCR. All mRNAs were detected except for ACE and ACE2. REN mRNA abundance was increased by cAMP and AZA (P < 0.0001). However, cAMP was more effective than AZA at both 24 and 48 h (P < 0.0001; P < 0.05) at the concentrations employed. AZA significantly increased AGT mRNA levels at 24 h (P = 0.001) and AGTR1 mRNA levels at 48 h (P < 0.0001), while cAMP had no effect. P TGS2 mRNA expression was increased by cAMP at 24 h (P < 0.0001) and by AZA at 48 h (P < 0.0001). cAMP and AZA had no effect on ATP6AP2 abundance. Thus, prorenin may function directly through the prorenin receptor, since cAMP upregulated both REN and its downstream target PTGS2 in HTR-8/SVneo cells, especially considering that ACE and ACE2 were not expressed. Our findings also suggest that expression of the RAS in trophoblast is regulated by DNA methylation as there was sustained over expression of REN, AGT, AGTR1 and PTGS2 with AZA. The ATP6AP2 promoter possesses CpG islands; however, its expression was not affected by AZA. This may suggest that either DNA methylation is not involved in the regulation of this gene, or that the over expressed prorenin binding to its receptor is causing suppression of ATP6AP2 abundance via a previously identified negative feedback pathway. (1) Pringle et al., Perinatal Society of Australia & New Zealand (PSANZ) 2010, Wellington, New Zealand (Abstract).(2) Itskovitz et al., Journal of Clinical Endocrinology and Metabolism, 1992, 75: 906–10.

Identification of vitamin D (VDR) and retinoic X (RXR) receptor in normal and neoplastic human reproductive tissues

2013 ◽  
Author(s):  
Federica Cariati ◽  
Vincenzo Gigantino ◽  
Giorgio Coppola ◽  
Claudia Pivonello ◽  
Mariano Galdiero ◽  
...  
Keyword(s):  
Vitamin D ◽  
Reproductive Tissues ◽  
Human Reproductive

scholarly journals Differential expression of TgMIC1 in isolates of Chinese 1 Toxoplasma with different virulence

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Yang Wang ◽  
Chengjian Han ◽  
Rongsheng zhou ◽  
Jinjin Zhu ◽  
Famin Zhang ◽  
...  
Keyword(s):  
Differential Expression ◽  
Rna Sequencing ◽  
Polyclonal Antibody ◽  
Protein Level ◽  
Polyclonal Antibodies ◽  
Mrna Levels ◽  
Quantitative Real Time Pcr ◽  
Sequencing Data ◽  
Predominant Genotype ◽  
High Virulence

Abstract Background The predominant genotype of Toxoplasma in China is the Chinese 1 (ToxoDB#9) lineage. TgCtwh3 and TgCtwh6 are two representative strains of Chinese 1, exhibiting high and low virulence to mice, respectively. Little is known regarding the virulence mechanism of this non-classical genotype. Our previous RNA sequencing data revealed differential mRNA levels of TgMIC1 in TgCtwh3 and TgCtwh6. We aim to further confirm the differential expression of TgMIC1 and its significance in this atypical genotype. Methods Quantitative real-time PCR was used to verify the RNA sequencing data; then, polyclonal antibodies against TgMIC1 were prepared and identified. Moreover, the invasion and proliferation of the parasite in HFF cells were observed after treatment with TgMIC1 polyclonal antibody or not. Results The data showed that the protein level of TgMIC1 was significantly higher in high-virulence strain TgCtwh3 than in low-virulence strain TgCtwh6 and that the invasion and proliferation of TgCtwh3 were inhibited by TgMIC1 polyclonal antibody. Conclusion Differential expression of TgMIC1 in TgCtwh3 and TgCtwh6 may explain, at least partly, the virulence mechanism of this atypical genotype.

Amniotic fluid LPCAT1 mRNA correlates with the lamellar body count

2016 ◽  
Vol 44 (5) ◽  
Author(s):  
Robert A. Welch ◽  
Michael K. Shaw ◽  
Kathryn C. Welch
Keyword(s):  
Amniotic Fluid ◽  
Pulmonary Surfactant ◽  
Short Communication ◽  
Lamellar Body ◽  
Future Research ◽  
Mrna Levels ◽  
Phospholipid Biosynthesis ◽  
Human Amniotic Fluid ◽  
Perinatal Medicine ◽  
Body Count

AbstractLysophosphatidylcholine acyltransferase 1 (LPCAT1) is required in the biosynthesis of pulmonary surfactant. This short communication describes our assessment of LPCAT1 mRNA levels in human amniotic fluid. We found a direct correlation between LPCAT1 mRNA copies and the amniotic fluid lamellar body count (LBC). This finding corroborates an association between LPCAT1 and surfactant phospholipid biosynthesis in humans. It may provide a model for future research in perinatal medicine.

Genome-wide screening of m6A-modified transcript in the Wilms’ tumor

2021 ◽  
Author(s):  
Zhuo Liu ◽  
Feng He ◽  
Jing Liu ◽  
Shengrong OuYang ◽  
Zexi Li ◽  
...  
Keyword(s):  
Gene Ontology ◽  
Real Time ◽  
Real Time Pcr ◽  
Wilms Tumor ◽  
Expression Patterns ◽  
Mrna Levels ◽  
Gene Ontology Analysis ◽  
Quantitative Real Time Pcr ◽  
Related Factors ◽  
Significant Difference

Abstract Background Wilms’ tumor, also called nephroblastoma, is the most common pediatric renal malignancy. The pathogenesis of Wilms’ tumor has been attributed to several genetic and epigenetic factors. However, the most pervasive internal mRNA modification that affects almost every process of RNA metabolism, RNA N6-Methyladenosine (m6A) methylation, has not been characterized in Wilms’ tumor. Methods Wilms’ tumor (WT) and adjacent non-cancerous (NC) tissue samples were obtained from 23 children with nephroblastoma, and the global m6A levels were measured by mass spectrometry. Analyses by m6A-mRNA epitranscriptomic microarray and mRNA microarray were performed, and m6A-related mRNAs were validated by quantitative real-time PCR for input and m6A-immunoprecipitated RNA samples from WT and NC tissues. Gene ontology analysis and KEGG pathway analysis were performed for differentially expressed genes, and expression of RNA methylation-related factors was measured by quantitative real-time PCR. Results The total m6A methylation levels in total RNA of WT samples and NC samples were (0.21 ± 0.01)% and (0.22 ± 0.01)%, respectively, with no statistically significant difference. Fifty-nine transcripts were differentially m6A-methylated between the WT and NC groups, which showed distinct m6A modification patterns. Gene ontology analysis indicated that m6A-modified genes were enriched in cancer-associated pathways, including the mTOR pathway, and conjoint analysis of the unique methylation and gene expression patterns in WT samples suggested an association with metabolic pathways.The mRNA levels of the m6A-related “reader” genes, YTHDF1, YTHDF2 and IGF2BP3, were statistically higher in WT samples than in NC samples. Conclusion This is the first study to determine the m6A modification profiles in Wilms’ tumor. Our data provide novel information regarding patterns of m6A modification that correlate with carcinogenesis in Wilms’ tumor.

scholarly journals Boden Black (A Novel) and With Axe and Pen in the New Zealand Alps: Differences Between Overseas and New Zealand Written Accounts of Climbing Mount Cook 1882-1920 and the Emergence of a New Zealand Voice in Mountaineering Literature

2021 ◽  
Author(s):  
◽  
Jura Fearnley
Keyword(s):  
New Zealand ◽  
Family Members ◽  
Southern Alps ◽  
Critical Study ◽  
First Person ◽  
The Novel ◽  
Writing Style ◽  
Natural Landscape ◽  
Site Specific ◽  
Mackenzie Basin

<p>This thesis has two components: creative and critical. The creative component is the novel Boden Black. It is a first person narrative, imagined as a memoir, and traces the life of its protagonist, Boden Black, from his childhood in the late 1930s to adulthood in the present day. The plot describes various significant encounters in the narrator’s life: from his introduction to the Mackenzie Basin and the Mount Cook region in the South Island of New Zealand, through to meetings with mountaineers and ‘lost’ family members. Throughout his journey from child to butcher to poet, Boden searches for ways to describe his response to the natural landscape. The critical study is titled With Axe and Pen in the New Zealand Alps. It examines the published writing of overseas and New Zealand mountaineers climbing at Aoraki/Mount Cook between 1882 and 1920. I advance the theory that there are stylistic differences between the writing of overseas and New Zealand mountaineers and that the beginning of a distinct New Zealand mountaineering voice can be traced back to the first accounts written by New Zealand mountaineers attempting to reach the summit of Aoraki/Mount Cook. The first mountaineer to attempt to climb Aoraki/Mount Cook was William Spotswood Green, an Irishman who introduced high alpine climbing to New Zealand in 1882. Early New Zealand mountaineers initially emulated the conventions of British mountaineering literature as exemplified by Green and other famous British mountaineers. These pioneering New Zealand mountaineers attempted to impose the language of the ‘civilised’ European alpine-world on to the ‘uncivilised’ world of the Southern Alps. However, as New Zealand mountaineering became more established at Aoraki/Mount Cook from the 1890s through to 1920, a distinct New Zealand voice developed in mountaineering literature: one that is marked by a sense of connection to place expressed through site-specific, factual observation and an unadorned, sometimes laconic, vernacular writing style.</p>

scholarly journals Cloning, Characterization, and Expression Features of Chicken CDS2 Splicing Variants

2020 ◽  
Author(s):  
Yuanyuan Xu ◽  
Shuping Zhang ◽  
Yujun Guo ◽  
Wen Chen ◽  
Yanqun Huang
Keyword(s):  
Adipose Tissue ◽  
Real Time ◽  
Real Time Pcr ◽  
Expression Patterns ◽  
Mrna Levels ◽  
Exogenous Insulin ◽  
Quantitative Real Time Pcr ◽  
Temporal Expression ◽  
Spatio Temporal ◽  
The Brain

Abstract Background: The CDS gene encodes the CDP-diacylglycerol synthase enzyme that catalyzes the formation of CDP-diacylglycerol (CDP-DAG) from phosphatidic acid. At present, there are no reports of CDS2 in birds. Here, we identified chicken CDS2 transcripts by combining conventional RT- PCR amplification, 5' RACE (Fig. 1A), and 3' RACE, explored the spatio-temporal expression profiles of total CDS2 and the longest transcript variant CDS2-4, and investigated the effect of exogenous insulin on total the mRNA level of CDS2 by quantitative real-time PCR. Results: Four transcripts of chicken CDS2 (CDS2-1, -2, -3, and -4) were identified, which were alternatively spliced at the 3′-untranslated region (UTR). CDS2 was widely expressed in all tissues examined and the longest variant CDS2-4 was the major transcript. Both total CDS2 and CDS2-4 were prominently expressed in adipose tissue and the heart, and exhibited low expression in the liver and pectoralis of 49 day-old chickens. Quantitative real-time PCR revealed that total CDS2 and CDS2-4 had different spatio-temporal expression patterns in chicken. Total CDS2 exhibited a similar temporal expression tendency with a high level in the later period of incubation (embryonic day 19 [E19] or 1-day-old) in the brain, liver, and pectoralis. While CDS2-4 presented a distinct temporal expression pattern in these tissues, CDS2-4 levels peaked at 21 days in the brain and pectoralis, while liver CDS2-4 mRNA levels were highest at the early stage of hatching (E10). Total CDS2 (P < 0.001) and CDS2-4 (P = 0.0090) mRNA levels in the liver were differentially regulated throughout development of the chicken. Exogenous insulin significantly downregulated the level of total CDS2 at 240 min in the pectoralis of Silky chickens (P < 0.01). Total CDS2 levels in the liver of Silky chickens were higher than that of the broiler in the basal state and after insulin stimulation. Conclusion: Chicken CDS2 has multiple transcripts with variation at the 3′-UTR, which was prominently expressed in adipose tissue. Total CDS2 and CDS2-4 presented distinct spatio-temporal expression patterns, and they were differentially regulated with age in liver. Insulin could regulate chicken CDS2 levels in a breed- and tissue-specific manner.

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