Ontogeny and the effects of corticosteroid pretreatment on aquaporin water channels in the ovine cerebral cortex

2005 ◽  
Vol 17 (5) ◽  
pp. 535 ◽  
Author(s):  
Nitin P. Ron ◽  
John A. Kazianis ◽  
James F. Padbury ◽  
Courtney M. Brown ◽  
Bethany G. McGonnigal ◽  
...  
Keyword(s):  
Cerebral Cortex ◽  
Protein Expression ◽  
Mrna Expression ◽  
Ribosomal Gene ◽  
Adult Brain ◽  
Aquaporin 4 ◽  
Adult Sheep ◽  
Mrna And Protein Expression ◽  
Northern Hybridisation ◽  
Aqp4 Protein

The aim of the present study was to determine the ontogeny and effects of corticosteroid pretreatment on aquaporin 4 (AQP4) channel mRNA and protein expression in the cerebral cortex of sheep during development. A portion of the cerebral cortex was snap-frozen from fetuses of dexamethasone- and placebo-treated ewes at 60%, 80% and 90% of gestation, dexamethasone- and placebo-treated newborn lambs and adult sheep. Cerebral cortical samples were obtained 18 h after the last of four 6 mg dexamethasone or placebo injections were given over 48 h to the ewes and adult sheep. Lambs were treated with 0.01 mg kg−1 dexamethasone or placebo in the same schedule as the ewes and adult sheep. Amplification of an ovine AQP4 cDNA fragment was accomplished by reverse transcription–polymerase chain reaction using primers based on a homologous bovine sequence. The resulting cDNA was used to determine AQP4 channel mRNA expression by Northern hybridisation using phosphorimaging. The relative abundance of AQP4 mRNA was normalised to the ovine ribosomal gene L32. A portion of the frontal cortex was also analysed for AQP4 protein expression by Western immunoblot. Densitometry was performed and the results expressed as a ratio to an adult brain pool. Aquaporin 4 channel mRNA and protein were detectable as early as at 60% gestation. There were no changes in AQP4 mRNA expression among the fetal, newborn and adult groups or after dexamethasone pretreatment in any age group. The expression of the AQP4 protein was higher (P < 0.05) in fetuses at 80% and 90% of gestation (2.9- and 3.3-fold, respectively), in lambs (3.2-fold) and in adult sheep (3.8-fold) compared with fetuses at 60% of gestation, as well as in adult sheep (1.3-fold) compared with fetuses at 80% of gestation. Dexamethasone pretreatment resulted in decreases (P < 0.05) in AQP4 protein expression in the lambs and adult sheep, but not in the fetal groups. We conclude that: (1) AQP4 mRNA and protein were expressed early in fetal and throughout ovine development; (2) protein, but not mRNA, expression increased between 60% and 80% of gestation and did not differ from adult levels by 90% of gestation; and (3) dexamethasone pretreatment resulted in decreases in AQP4 protein expression in lambs and adult sheep, but not in fetuses. The maturational increases in AQP4 protein expression and dexamethasone-related decreases in expression were post-transcriptional, because changes in AQP4 mRNA expression were not observed.

scholarly journals Differential between Protein and mRNA Expression of CCR7 and SSTR5 Receptors in Crohn's Disease Patients

2009 ◽  
Vol 2009 ◽  
pp. 1-9 ◽  
Author(s):  
Nathalie Taquet ◽  
Serge Dumont ◽  
Jean-Luc Vonesch ◽  
Didier Hentsch ◽  
Jean-Marie Reimund ◽  
...  
Keyword(s):  
Crohn’S Disease ◽  
Crohn's Disease ◽  
Protein Expression ◽  
Mrna Expression ◽  
Large Scale ◽  
Mononuclear Cells ◽  
Biologically Active ◽  
Chronic Inflammatory Bowel Disease ◽  
Peripheral Blood Mononuclear ◽  
Mrna And Protein Expression

Crohn's disease (CD) is a multifactorial chronic inflammatory bowel disease of unknown cause. The aim of the present study was to explore if mRNA over-expression of SSTR5 and CCR7 found in CD patients could be correlated to respective protein expression. When compared to healthy donors, SSTR5 was over-expressed 417±71 times in CD peripheral blood mononuclear cells (PBMCs). Flow cytometry experiments showed no correlation between mRNA and protein expression for SSTR5 in PBMCs. In an attempt to find a reason of such a high mRNA expression, SSTR5 present on CD PBMCs were tested and found as biologically active as on healthy cells. In biopsies of CD intestinal tissue, SSTR5 was not over-expressed but CCR7, unchanged in PBMCs, was over-expressed by 10±3 times in the lamina propria. Confocal microscopy showed a good correlation of CCR7 mRNA and protein expression in CD intestinal biopsies. Our data emphasize flow and image cytometry as impossible to circumvent in complement to molecular biology so to avoid false interpretation on receptor expressions. Once confirmed by further large-scale studies, our preliminary results suggest a role for SSTR5 and CCR7 in CD pathogenesis.

Parathyroid hormone stimulates endothelial expression of atherosclerotic parameters through protein kinase pathways

2007 ◽  
Vol 292 (4) ◽  
pp. F1215-F1218 ◽  
Author(s):  
Gloria Rashid ◽  
Jacques Bernheim ◽  
Janice Green ◽  
Sydney Benchetrit
Keyword(s):  
Parathyroid Hormone ◽  
Protein Kinase ◽  
Protein Expression ◽  
Mrna Expression ◽  
Umbilical Vein ◽  
Protein Levels ◽  
Glycation End Products ◽  
Mrna And Protein Expression ◽  
Vascular Structures ◽  
The Impact

Parathyroid hormone (PTH), the major systemic calcium-regulating hormone, has been linked to uremic vascular changes. Considering the possible deleterious action of PTH on vascular structures, it seemed logical to evaluate the impact of PTH on the receptor of advanced glycation end products (RAGE) and interleukin 6 (IL-6) mRNA and protein expression, taking into account that such parameters might be involved in the pathogenesis of vascular calcification, atherosclerosis, and/or arteriolosclerosis. Human umbilical vein cord endothelial cells (HUVEC) were stimulated for 24 h with 10−12–10−10 mol/l PTH. The mRNA expression of RAGE and IL-6 was established by reverse transcriptase/PCR techniques. RAGE protein levels were determined by Western blot and IL-6 secretion was measured by ELISA. The pathways by which PTH may have an effect on HUVEC functions were evaluated. PTH (10−11–10−10mol/l) significantly increased RAGE mRNA and protein expression. PTH also significantly increased IL-6 mRNA expression without changes at protein levels. The addition of protein kinase (PKC or PKA) inhibitors or nitric oxide (NO) synthase inhibitors significantly reduced the RAGE and IL-6 mRNA expression and the RAGE protein expression. PTH stimulates the mRNA expressions of RAGE and IL-6 and the protein expression of RAGE. These stimulatory effects are probably through PKC and PKA pathways and are also NO dependent. Such data may explain the possible impact of PTH on the atherosclerotic and arteriosclerotic progression.

scholarly journals Expression of ACE2 in the Intact and Acutely Injured Kidney

Kidney360 ◽  
2021 ◽  
pp. 10.34067/KID.0001562021
Author(s):  
Karl A. Nath ◽  
Raman Deep Singh ◽  
Joseph P. Grande ◽  
Vesna D. Garovic ◽  
Anthony J. Croatt ◽  
...  
Keyword(s):  
Protein Expression ◽  
Mrna Expression ◽  
Heme Oxygenase ◽  
Urinary Tract Obstruction ◽  
Kidney Injury ◽  
Heme Oxygenase 1 ◽  
Renal Tubular Epithelium ◽  
Mrna And Protein Expression ◽  
Intact Kidney ◽  
Renal Tubular

Background. The actions of angiotensin-converting enzyme 2 (ACE2) oppose those of the renin-angiotensin-aldosterone system. Evidence supports ACE2 as a cytoprotectant in some tissues. This study examined ACE2 expression in models of acute kidney injury (AKI). Methods. ACE2 mRNA and protein expression, ACE2 activity, and ACE2 expression by immunofluorescence were assessed following ischemic AKI in mice. Renal ACE2 mRNA expression was evaluated in lipopolysaccharide-induced AKI in wildtype (C57BL/6J) mice, in heme oxygenase-1+/+ and heme oxygenase-1-/- mice, and following unilateral urinary tract obstruction (UUO) in wildtype mice. The effect of sex and age on renal ACE2 protein expression was also assessed. Results. In ischemic AKI, ACE2 mRNA and protein expression and ACE2 activity were reduced as compared with such indices in the intact kidney. In ischemic AKI, ACE2, which, in health, is prominently expressed in the renal tubular epithelium, especially in proximal tubules, exhibited decreased expression in these segments. Decreased ACE2 expression in AKI did not reflect reduced GFR per se as ACE2 mRNA expression was unaltered after UUO. Lipopolysaccharide induced renal ACE2 mRNA expression in wildtype mice, but this effect of lipopolysaccharide did not occur in heme oxygenase-1 deficient mice. In the intact kidney, renal ACE2 protein expression decreased in female mice as compared with male mice, but was unaltered with age. Conclusion. We conclude that renal ACE2 expression is decreased in ischemic AKI, one characterized by markedly reduced GFR and abundant cell death, but is upregulated in lipopolysaccharide-induced AKI; this latter effect requires heme oxygenase-1. Determining the significance of ACE2 expression in models of AKI merits further study. We also suggest that understanding the mechanism underlying ACE2 downregulation in AKI may offer insights relevant to COVID-19: ACE2 is downregulated after ACE2 mediates SARS-CoV-2 cellular entry; such downregulation promotes inflammation in COVID-19; and AKI commonly occurs and determines outcomes in COVID-19.

Wnt/β-Catenin and Hippo Pathway Deregulation in Mammary Tumors of Humans, Dogs, and Cats

2020 ◽  
Vol 57 (6) ◽  
pp. 774-790
Author(s):  
Alessandro Sammarco ◽  
Chiara Gomiero ◽  
Roberta Sacchetto ◽  
Giorgia Beffagna ◽  
Silvia Michieletto ◽  
...  
Keyword(s):  
Protein Expression ◽  
Mrna Expression ◽  
Quantitative Polymerase Chain Reaction ◽  
Mrna Level ◽  
Hippo Pathway ◽  
Mammary Tumors ◽  
Breast Cancers ◽  
Tumor Tissues ◽  
Mrna And Protein Expression ◽  
Polymerase Chain

Mammary cancer is a common neoplasm in women, dogs, and cats that still represents a therapeutic challenge. Wnt/β-catenin and Hippo pathways are involved in tumor progression, cell differentiation, and metastasis. The aim of this study was to evaluate mRNA and protein expression of molecules involved in these pathways in human (HBC), canine (CMT), and feline mammary tumors (FMT). Real-time quantitative polymerase chain reaction (qPCR) for β-catenin, CCND1, YAP, TAZ, CTGF, and ANKRD1, western blotting for YAP, TAZ, and β-catenin, and immunohistochemistry for estrogen receptor (ER), progesterone receptor (PR), ERBB2, β-catenin, and YAP/TAZ were performed on mammary tumor tissues. The protein expression of active β-catenin was higher in tumors than in healthy tissues in all 3 species. The mRNA expression of the downstream gene CCND1 was increased in HBC ER+ and CMTs compared to healthy tissues. Membranous and cytoplasmic protein expression of β-catenin were strongly negatively correlated in all 3 species. Tumors showed an increased protein expression of YAP/TAZ when compared to healthy tissues. Notably, YAP/TAZ expression was higher in triple negative breast cancers when compared to HBC ER+ and in FMTs when compared to CMTs. The mRNA expression of β-catenin, YAP, TAZ, CTGF, and ANKRD1 was not different between tumors and healthy mammary gland in the 3 species. This study demonstrates deregulation of Wnt/β-catenin and Hippo pathways in mammary tumors, which was more evident at the protein rather than the mRNA level. Wnt/β-catenin and Hippo pathways seem to be involved in mammary carcinogenesis and therefore represent interesting therapeutic targets that should be further investigated.

Creatine kinase isoenzymes are highly regulated during pregnancy in rat uterus and placenta

1993 ◽  
Vol 265 (4) ◽  
pp. E624-E635 ◽  
Author(s):  
R. M. Payne ◽  
D. L. Friedman ◽  
J. W. Grant ◽  
M. B. Perryman ◽  
A. W. Strauss
Keyword(s):  
Creatine Kinase ◽  
Protein Expression ◽  
Mrna Expression ◽  
Posttranscriptional Regulation ◽  
Nuclear Gene ◽  
Rat Uterus ◽  
Pregnant Rats ◽  
Pregnancy And Delivery ◽  
Mrna And Protein Expression

Creatine kinase (CK) isoenzymes play a central role in energy transfer. Expression of CK isoenzymes in rat uterus and placenta was analyzed to evaluate their contribution to energy metabolism during pregnancy and delivery. Tissue from the uterine horns and placentas of pregnant rats from day 14 of gestation to 17 days postpartum was analyzed for expression of “brain” CK (BCK) and ubiquitous mitochondrial CK (uMtCK) mRNA, protein, and enzyme activity. uMtCK mRNA expression is high in prepartum uterus, but rapidly falls (> 10-fold) after delivery to a nadir at 7 days postpartum. Prepartum BCK mRNA expression is coordinate with uMtCK but has a 15-fold greater expression than uMtCK. Both CK mRNAs rise by 17 days postpartum. Both BCK and uMtCK mRNA expressions are strongly induced in placenta at 20 days gestation with a rapid fall (> 6-fold) immediately before delivery. Protein expression of BCK and uMtCK is also coordinate. However, analysis of mRNA and protein expression indicates that significant posttranscriptional regulation of both kinds of CK also occurs. CK activity in uterus and placenta reflects BCK expression. By immunohistochemistry, BCK and uMtCK protein expression is highly localized in the placenta and endometrium of prepartum uterus. This expression shifts entirely to the uterine smooth muscle by 17 days postpartum. uMtCK mRNA expression is rapidly induced by beta-estradiol in vitro (> 6-fold), demonstrating estrogen-responsive elements in the uMtCK nuclear gene. Thus a second isoenzyme of CK, uMtCK, is expressed in rat uterus and placenta and is highly regulated with BCK. These results suggest an important role for CK in the maintenance and termination of pregnancy.

Regulation of α7-integrin expression in vascular smooth muscle by injury-induced atherosclerosis

2004 ◽  
Vol 287 (1) ◽  
pp. H381-H389 ◽  
Author(s):  
Jun-Tzu Chao ◽  
Gerald A. Meininger ◽  
Jan L. Patterson ◽  
Sarah A. L. Jones ◽  
Charles R. Partridge ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Protein Expression ◽  
Mrna Expression ◽  
Vascular Injury ◽  
Pyrrolidine Dithiocarbamate ◽  
Integrin Subunit ◽  
Integrin Expression ◽  
Mrna Levels ◽  
Mrna And Protein Expression

Injury of vascular smooth muscle cells (VSMCs) by allylamine (AAM) leads to phenotypic changes associated with atherogenic progression including increased proliferation, migration, and alterations in cell adhesion. In the present study, the relationship between AAM-induced vascular injury and expression of the α7-integrin subunit was investigated. The α7-mRNA and protein expression were examined using real-time RT-PCR, fluorescence-activated cell sorting analysis (FACS), immunohistochemistry, and immunoblotting. In cultured VSMCs from aortas of AAM-treated rats (70 mg/kg for 20 days), α7-mRNA levels were increased more than twofold compared with control cells. No change was seen in β1-integrin expression. FACS analysis revealed increased cell surface expression of α7-protein (25 ± 9%; * P < 0.05). AAM treatment of naive VSMCs enhanced α7-mRNA expression (2.4 ± 0.7-fold, mean ± SE; * P < 0.05). The increased α7-mRNA expression was attenuated by the amine oxidase inhibitor semicarbazide and the antioxidant pyrrolidine dithiocarbamate, which confirms a role for oxidative stress in modulating α7-expression. In vivo α7-mRNA and protein expression were enhanced in the aortas of AAM-treated rats. In addition, increased α7-integrin expression facilitated AAM VSMC adhesion to laminin more efficiently compared with control (51 ± 2%; * P < 0.05). Chemical injury induced by AAM significantly enhances α7-integrin expression in VSMCs. These findings implicate for the first time the expression of α7-integrin during the response of VSMCs to vascular injury.

The Expression of FAT1 is Associated with Overall Survival in Children with Medulloblastoma

2016 ◽  
Vol 103 (1) ◽  
pp. 44-52 ◽  
Author(s):  
Jianzhong Yu ◽  
Hao Li
Keyword(s):  
Overall Survival ◽  
Protein Expression ◽  
Wnt Signaling ◽  
Mrna Expression ◽  
Wnt Signaling Pathway ◽  
Rank Test ◽  
Missense Mutations ◽  
Kaplan Meier ◽  
Whole Exome ◽  
Mrna And Protein Expression

Purpose The FAT1 gene is involved in some cancers; however, its role in medulloblastoma is less clear. This study investigated the effects of FAT1 expression on the prognosis of medulloblastoma patients. Methods Whole exome sequencing was undertaken in 40 medulloblastoma patient samples. FAT1 mRNA and protein expression levels in normal and brain tumor tissues were determined by fluorescence quantitative PCR and immunohistochemistry, respectively. The association of FAT1 expression with overall survival (OS) was examined by Kaplan-Meier curve analysis with a log-rank test. Following lentiviral-mediated FAT1 knockdown using shRNA in Daoy cells, proliferation, Wnt signaling, and β-catenin protein expression were determined. Results Eight FAT1 missense mutations were detected in 7 patients. FAT1 mRNA expression in tumors was significantly lower than in adjacent normal tissue ( p = 0.043). The OS of patients with high FAT1 protein expression was significantly longer than that of patients with low FAT1 protein expression (median survival time: 24.3 vs 4.8 months, respectively; p = 0.002). shFAT1 cells had significantly higher proliferation rates than shControl cells ( p≤0.028). Furthermore, the mRNA expression of LEF1, β-catenin, and cyclin D1 was significantly upregulated in shFAT1-Daoy cells ( p≤0.018). Conclusions Low FAT1 expression was associated with poor prognosis in children with medulloblastoma. Furthermore, FAT1 may act on Wnt signaling pathway to exert its antitumor effect.

scholarly journals High Expression of LTBP2 Contributes to Poor Prognosis in Colorectal Cancer Patients and Correlates with the Mesenchymal Colorectal Cancer Subtype

2019 ◽  
Vol 2019 ◽  
pp. 1-9 ◽  
Author(s):  
Ying Huang ◽  
Guihua Wang ◽  
Chunmei Zhao ◽  
Rong Geng ◽  
Shu Zhang ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Overall Survival ◽  
Protein Expression ◽  
Mrna Expression ◽  
Cell Lines ◽  
Critical Role ◽  
Prognostic Significance ◽  
Poor Overall Survival ◽  
Expression Levels ◽  
Mrna And Protein Expression

Colorectal cancer (CRC) is a complex and heterogeneous disease with four consensus molecular subtypes (CMS1-4). LTBP2 is a member of the fibrillin/LTBP super family and plays a critical role in tumorigenesis by activating TGF-β in the CMS4 CRC subtype. So far, the expression and prognostic significance of LTBP2 in CRC remains obscure. In this study, we aimed to analyze the mRNA and protein expression levels of LTBP2 in CRC tissues and then estimate their values as a potential prognostic biomarker. We detected the mRNA expression of LTBP2 in 28 cases of fresh CRC tissues and 4 CRC cell lines and the protein expression of LTBP2 in 483 samples of CRC tissues, matched tumor-adjacent tissues, and benign colorectal diseases. LTBP2 protein expression was then correlated to patients’ clinical features and overall survival. Both LTBP2 mRNA and protein expression levels in CRC tissues were remarkably superior to those in adjacent normal colorectal tissues (P=0.0071 and P<0.001, respectively), according to TCGA dataset of CRC. High LTBP2 protein expression was correlated with TNM stage (P<0.001), T stage (P<0.001), N stage (P<0.001), and M stage (P<0.001). High LTBP2 protein expression was related to poor overall survival in CRC patients and was an independent prognostic factor for CRC. LTBP2 mRNA expression was especially higher in the CMS4 subtype (P<0.001), which was confirmed in CRC cell lines. Our data suggested that LTBP2 may act as an oncogene in the development of colorectal cancer and have important significance in predicting CRC prognosis. LTBP2 could be a novel biomarker and potential therapeutic target for mesenchymal colorectal cancer and can improve the outcome of high-risk CRC.

scholarly journals Increased Tau Expression Correlates With IDH Mutation in Infiltrating Gliomas and Impairs Cell Migration

2020 ◽  
Vol 79 (5) ◽  
pp. 493-499
Author(s):  
Satoshi Nakata ◽  
Antionette Price ◽  
Charles Eberhart ◽  
Meaghan Morris
Keyword(s):  
Cell Migration ◽  
Protein Expression ◽  
Mrna Expression ◽  
Tumor Invasion ◽  
Tissue Microarrays ◽  
Brain Diseases ◽  
Idh Mutation ◽  
Mrna And Protein Expression ◽  
Patient Prognosis ◽  
Tau Expression

Abstract Although the microtubule-associated protein tau is well studied in human neurodegeneration, the role of tau in neoplastic brain diseases is not well understood. Recently, studies have shown tau mRNA expression is associated with improved survival in human infiltrating gliomas. However, the biologic basis of this association is largely unexplored. Using 2 independent publicly available mRNA databases, we show that high tau mRNA expression is associated with improved patient survival in infiltrating gliomas. Higher tau protein expression is also associated with improved patient prognosis in infiltrating gliomas by immunohistochemical staining of tissue microarrays. This prognostic association is in part due to higher tau mRNA and protein expression in IDH-mutant infiltrating astrocytomas. Expression of tau in an IDH-wildtype glioblastoma cell line selectively impairs cell migration in assays designed to mimic tumor invasion. These findings suggest that tau expression is not only associated with IDH mutation status but also may contribute to improved patient outcomes by impairing tumor invasion.

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