scholarly journals Evaluation of endogenous miRNA reference genes across different zebrafish strains, developmental stages and kidney disease models

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Florian Siegerist ◽  
Tim Lange ◽  
Anna Iervolino ◽  
Thor Magnus Koppe ◽  
Weibin Zhou ◽  
...  
Keyword(s):  
Kidney Disease ◽  
Reference Genes ◽  
Developmental Stages ◽  
Kidney Diseases ◽  
Morphological Changes ◽  
Group Differences ◽  
Disease Models ◽  
Endogenous Reference ◽  
Process Architecture

AbstractThe majority of kidney diseases arise from the loss of podocytes and from morphological changes of their highly complex foot process architecture, which inevitably leads to a reduced kidney filtration and total loss of kidney function. It could have been shown that microRNAs (miRs) play a pivotal role in the pathogenesis of podocyte-associated kidney diseases. Due to their fully functioning pronephric kidney, larval zebrafish have become a popular vertebrate model, to study kidney diseases in vivo. Unfortunately, there is no consensus about a proper normalization strategy of RT-qPCR-based miRNA expression data in zebrafish. In this study we analyzed 9 preselected candidates dre-miR-92a-3p, dre-miR-206-3p, dre-miR-99-1, dre-miR-92b-3p, dre-miR-363-3p, dre-let-7e, dre-miR-454a, dre-miR-30c-5p, dre-miR-126a-5p for their capability as endogenous reference genes in zebrafish experiments. Expression levels of potential candidates were measured in 3 different zebrafish strains, different developmental stages, and in different kidney disease models by RT-qPCR. Expression values were analyzed with NormFinder, BestKeeper, GeNorm, and DeltaCt and were tested for inter-group differences. All candidates show an abundant expression throughout all samples and relatively high stability. The most stable candidate without significant inter-group differences was dre-miR-92b-3p making it a suitable endogenous reference gene for RT-qPCR-based miR expression zebrafish studies.

scholarly journals Evaluation of endogenous miRNA reference genes across different zebrafish strains, developmental stages and kidney disease models

2021 ◽  
Author(s):  
Florian Siegerist ◽  
Tim Lange ◽  
Anna Iervolino ◽  
Thor Magnus Koppe ◽  
Weibin Zhou ◽  
...  
Keyword(s):  
Kidney Disease ◽  
Reference Genes ◽  
Developmental Stages ◽  
Kidney Diseases ◽  
Morphological Changes ◽  
Group Differences ◽  
Disease Models ◽  
Endogenous Reference ◽  
Process Architecture

The majority of kidney diseases arise from the loss of podocytes and from morphological changes of their highly complex foot process architecture, which inevitably leads to a reduced kidney filtration and total loss of kidney function. It could have been shown that microRNAs (miRs) play a pivotal role in the pathogenesis of podocyte-associated kidney diseases. Due to their fully functioning pronephric kidney, larval zebrafish have become a popular vertebrate model, to study kidney diseases in vivo. Unfortunately, there is no consensus about a proper normalization strategy of RT-qPCR based miRNA expression data in zebrafish. In this study we analyzed 9 preselected candidates dre-miR-92a-3p, dre-miR-206-3p, dre-miR-99-1, dre-miR-92b-3p, dre-miR-363-3p, dre-let-7e, dre-miR-454a, dre-miR-30c-5p, dre-miR-126a-5p for their capability as endogenous reference genes in zebrafish experiments. Expression levels of potential candidates were measured in 3 different zebrafish strains, different developmental stages and in different kidney disease models by RT-qPCR. Expression values were analyzed with NormFinder, BestKeeper, GeNorm, and DeltaCt and were tested for inter-group differences. All candidates show an abundant expression throughout all samples and relatively high stability. The most stable candidate without significant inter-group differences was dre-miR-92b-3p making it a suitable endogenous reference gene for RT-qPCR-based miR expression zebrafish studies. The majority of kidney diseases arise from the loss of podocytes and from morphological changes of their highly complex foot process architecture, which inevitably leads to a reduced kidney filtration and total loss of kidney function. It could have been shown that microRNAs (miRs) play a pivotal role in the pathogenesis of podocyte-associated kidney diseases. Due to their fully functioning pronephric kidney, larval zebrafish have become a popular vertebrate model, to study kidney diseases in vivo. Unfortunately, there is no consensus about a proper normalization strategy of RT-qPCR-based miRNA expression data in zebrafish. In this study we analyzed 9 preselected candidates dre-miR-92a-3p, dre-miR-206-3p, dre-miR-99-1, dre-miR-92b-3p, dre-miR-363-3p, dre-let-7e, dre-miR-454a, dre-miR-30c-5p, dre-miR-126a-5p for their capability as endogenous reference genes in zebrafish experiments. Expression levels of potential candidates were measured in 3 different zebrafish strains, different developmental stages, and in different kidney disease models by RT-qPCR. Expression values were analyzed with NormFinder, BestKeeper, GeNorm, and DeltaCt and were tested for inter-group differences. All candidates show an abundant expression throughout all samples and relatively high stability. The most stable candidate without significant inter-group differences was dre-miR-92b-3p making it a suitable endogenous reference gene for RT-qPCR-based miR expression zebrafish studies.

scholarly journals Role of SIRT1 in HIV-associated kidney disease

2020 ◽  
Vol 319 (2) ◽  
pp. F335-F344 ◽  
Author(s):  
Xuan Wang ◽  
Ruijie Liu ◽  
Weijia Zhang ◽  
Deborah P. Hyink ◽  
Gokul C. Das ◽  
...  
Keyword(s):  
Kidney Disease ◽  
Drug Targets ◽  
Cell Injury ◽  
Kidney Diseases ◽  
Sirtuin 1 ◽  
Kidney Cells ◽  
Sirt1 Expression ◽  
Hiv 1

Human immunodeficiency virus (HIV) infection of kidney cells can lead to HIV-associated nephropathy (HIVAN) and aggravate the progression of other chronic kidney diseases. Thus, a better understanding of the mechanisms of HIV-induced kidney cell injury is needed for effective therapy against HIV-induced kidney disease progression. We have previously shown that the acetylation and activation of key inflammatory regulators, NF-κB p65 and STAT3, were increased in HIVAN kidneys. Here, we demonstrate the key role of sirtuin 1 (SIRT1) deacetylase in the regulation of NF-κB and STAT3 activity in HIVAN. We found that SIRT1 expression was reduced in the glomeruli of human and mouse HIVAN kidneys and that HIV-1 gene expression was associated with reduced SIRT1 expression and increased acetylation of NF-κB p65 and STAT3 in cultured podocytes. Interestingly, SIRT1 overexpression, in turn, reduced the expression of negative regulatory factor in podocytes stably expressing HIV-1 proviral genes, which was associated with inactivation of NF-κB p65 and a reduction in HIV-1 long terminal repeat promoter activity. In vivo, the administration of the small-molecule SIRT1 agonist BF175 or inducible overexpression of SIRT1 specifically in podocytes markedly attenuated albuminuria, kidney lesions, and expression of inflammatory markers in Tg26 mice. Finally, we showed that the reduction in SIRT1 expression by HIV-1 is in part mediated through miR-34a expression. Together, our data provide a new mechanism of SIRT1 regulation and its downstream effects in HIV-1-infected kidney cells and indicate that SIRT1/miR-34a are potential drug targets to treat HIV-related kidney disease.

scholarly journals Sonographic assessment of thyroid gland in patients with chronic kidney disease undergoing hemodialysis

2017 ◽  
Vol 5 (1) ◽  
pp. 4 ◽  
Author(s):  
Maisa Elzakil ◽  
Awadia Gareeballah ◽  
Moawia Gameraddin ◽  
Mowada Burai ◽  
Fathelrehman Alagab
Keyword(s):  
Chronic Kidney Disease ◽  
Thyroid Gland ◽  
Kidney Disease ◽  
Thyroid Nodules ◽  
Kidney Diseases ◽  
Morphological Changes ◽  
Thyroid Tissue ◽  
Thyroid Volume ◽  
Cross Sectional Study ◽  
Cross Sectional

Background: Thyroid gland morphology and disorders were affected with chronic kidney diseases (CKD). The study aims to assess thyroid gland disorders and morphology in patients with CKD on regular hemodialysis.Materials and methods: A cross-sectional study included 71 participants divided into two groups. The study group included 51 patients with known chronic kidney disease on hemodialysis and healthy group included 20 participants. The exclusion criteria were thyroid disorders. The thyroid gland was scanned with ultrasound using a 7 MHz probe.Results: The thyroid is enlarged in 21.57% of patients and heterogeneous echotexture in 31.4%. The prevalence of thyroid nodules and cysts were 9.8% and 7.8% respectively. A positive linear correlation existed between duration of hemodialysis and thyroid volume. Thyroid volume and echotexture were significantly increased with duration of hemodialysis (p = .001 and .00 respectively). Thyroid nodules and cysts were not significantly correlated with duration of hemodialysis (p = .06 and .28 respectively).Conclusions: In conclusion, enlargement of thyroid gland and heterogeneity of thyroid tissue were the most common morphological changes in patients with chronic renal failure undergoing hemodialysis. The prevalence increased with longer duration of hemodialysis. Periodic ultrasound assessment of thyroid volume and texture are recommended in hemodialyzed patients to avoid complications.

scholarly journals Targeting STAT3 signaling in kidney disease

2019 ◽  
Vol 316 (6) ◽  
pp. F1151-F1161 ◽  
Author(s):  
Jesse Pace ◽  
Praharshasai Paladugu ◽  
Bhaskar Das ◽  
John C. He ◽  
Sandeep K. Mallipattu
Keyword(s):  
Kidney Disease ◽  
Kidney Diseases ◽  
Janus Kinase ◽  
Disease Models ◽  
Pharmacological Inhibition ◽  
Stat3 Signaling ◽  
Stat Signaling ◽  
Progression Of Kidney Disease ◽  
Central Member

The Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway is a multifaceted transduction system that regulates cellular responses to incoming signaling ligands. STAT3 is a central member of the JAK/STAT signaling cascade and has long been recognized for its increased transcriptional activity in cancers and autoimmune disorders but has only recently been in the spotlight for its role in the progression of kidney disease. Although genetic knockout and manipulation studies have demonstrated the salutary benefits of inhibiting STAT3 activity in several kidney disease models, pharmacological inhibition has yet to make it to the clinical forefront. In recent years, significant effort has been aimed at suppressing STAT3 activation for treatment of cancers, which has led to the development of a wide variety of STAT3 inhibitors, but only a handful have been tested in kidney disease models. Here, we review the detrimental role of dysregulated STAT3 activation in a variety of kidney diseases and the current progress in the treatment of kidney diseases with pharmacological inhibition of STAT3 activity.

scholarly journals Computational Modeling of Blood Flow Hemodynamics for Biomechanical Investigation of Cardiac Development and Disease

2021 ◽  
Vol 8 (2) ◽  
pp. 14
Author(s):  
Huseyin Enes Salman ◽  
Huseyin Cagatay Yalcin
Keyword(s):  
Heart Valves ◽  
Developmental Stages ◽  
Cardiac Development ◽  
Morphological Changes ◽  
Heart Defects ◽  
Animal Experiments ◽  
Cfd Modeling ◽  
Cfd Models ◽  
Developing Heart

The heart is the first functional organ in a developing embryo. Cardiac development continues throughout developmental stages while the heart goes through a serious of drastic morphological changes. Previous animal experiments as well as clinical observations showed that disturbed hemodynamics interfere with the development of the heart and leads to the formation of a variety of defects in heart valves, heart chambers, and blood vessels, suggesting that hemodynamics is a governing factor for cardiogenesis, and disturbed hemodynamics is an important source of congenital heart defects. Therefore, there is an interest to image and quantify the flowing blood through a developing heart. Flow measurement in embryonic fetal heart can be performed using advanced techniques such as magnetic resonance imaging (MRI) or echocardiography. Computational fluid dynamics (CFD) modeling is another approach especially useful when the other imaging modalities are not available and in-depth flow assessment is needed. The approach is based on numerically solving relevant physical equations to approximate the flow hemodynamics and tissue behavior. This approach is becoming widely adapted to simulate cardiac flows during the embryonic development. While there are few studies for human fetal cardiac flows, many groups used zebrafish and chicken embryos as useful models for elucidating normal and diseased cardiogenesis. In this paper, we explain the major steps to generate CFD models for simulating cardiac hemodynamics in vivo and summarize the latest findings on chicken and zebrafish embryos as well as human fetal hearts.

scholarly journals SHROOM3, the gene associated with chronic kidney disease, affects the podocyte structure

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Ryo Matsuura ◽  
Atsuko Hiraishi ◽  
Lawrence B. Holzman ◽  
Hiroki Hanayama ◽  
Koji Harano ◽  
...  
Keyword(s):  
Chronic Kidney Disease ◽  
Kidney Disease ◽  
Kidney Function ◽  
Genome Wide Association Study ◽  
Kidney Diseases ◽  
Vitro Experiment ◽  
Public Health Burden ◽  
In Vitro Experiment

AbstractChronic kidney disease is a public health burden and it remains unknown which genetic loci are associated with kidney function in the Japanese population, our genome-wide association study using the Biobank Japan dataset (excluding secondary kidney diseases, such as diabetes mellitus) clearly revealed that almost half of the top 50 single nucleotide polymorphisms associated with estimated glomerular filtration rate are located in the SHROOM3 gene, suggesting that SHROOM3 will be responsible for kidney function. Thus, to confirm this finding, supportive functional analyses were performed on Shroom3 in mice using fullerene-based siRNA delivery, which demonstrated that Shroom3 knockdown led to albuminuria and podocyte foot process effacement. The in vitro experiment shows that knockdown of Shroom3 caused defective formation of lamellipodia in podocyte, which would lead to the disruption of slit diaphragm. These results from the GWAS, in vivo and in vitro experiment were consistent with recent studies reporting that albuminuria leads to impairment of kidney function.

scholarly journals Lamin-B1 contributes to the proper timing of epicardial cell migration and function during embryonic heart development

2016 ◽  
Vol 27 (25) ◽  
pp. 3956-3963 ◽  
Author(s):  
Joseph R. Tran ◽  
Xiaobin Zheng ◽  
Yixian Zheng
Keyword(s):  
Cell Migration ◽  
Heart Development ◽  
Developmental Stages ◽  
Morphological Changes ◽  
Nuclear Shape ◽  
Epicardial Cells ◽  
Biological Phenomena ◽  
And Migration

Lamin proteins form a meshwork beneath the nuclear envelope and contribute to many different cellular processes. Mutations in lamins cause defective organogenesis in mouse models and human diseases that affect adipose tissue, brain, skeletal muscle, and the heart. In vitro cell culture studies have shown that lamins help maintain nuclear shape and facilitate cell migration. However, whether these defects contribute to improper tissue building in vivo requires further clarification. By studying the heart epicardium during embryogenesis, we show that Lb1-null epicardial cells exhibit in vivo and in vitro migratory delay. Transcriptome analyses of these cells suggest that Lb1 influences the expression of cell adhesion genes, which could affect cell migration during epicardium development. These epicardial defects are consistent with incomplete development of both vascular smooth muscle and compact myocardium at later developmental stages in Lb1-null embryos. Further, we found that Lb1-null epicardial cells have a delayed nuclear morphology change in vivo, suggesting that Lb1 facilitates morphological changes associated with migration. These findings suggest that Lb1 contributes to nuclear shape maintenance and migration of epicardial cells and highlights the use of these cells for in vitro and in vivo study of these classic cell biological phenomena.

scholarly journals Disease Modeling To Understand the Pathomechanisms of Human Genetic Kidney Disorders

2020 ◽  
Vol 15 (6) ◽  
pp. 855-872 ◽  
Author(s):  
Elisa Molinari ◽  
John A. Sayer
Keyword(s):  
Kidney Disease ◽  
Disease Modeling ◽  
Kidney Diseases ◽  
Model Systems ◽  
Definitive Treatment ◽  
Patient Specific ◽  
Disease Models ◽  
Culture Methods ◽  
Human Patient ◽  
Inherited Kidney Disease

The class of human genetic kidney diseases is extremely broad and heterogeneous. Accordingly, the range of associated disease phenotypes is highly variable. Many children and adults affected by inherited kidney disease will progress to ESKD at some point in life. Extensive research has been performed on various different disease models to investigate the underlying causes of genetic kidney disease and to identify disease mechanisms that are amenable to therapy. We review some of the research highlights that, by modeling inherited kidney disease, contributed to a better understanding of the underlying pathomechanisms, leading to the identification of novel genetic causes, new therapeutic targets, and to the development of new treatments. We also discuss how the implementation of more efficient genome-editing techniques and tissue-culture methods for kidney research is providing us with personalized models for a precision-medicine approach that takes into account the specificities of the patient and the underlying disease. We focus on the most common model systems used in kidney research and discuss how, according to their specific features, they can differentially contribute to biomedical research. Unfortunately, no definitive treatment exists for most inherited kidney disorders, warranting further exploitation of the existing disease models, as well as the implementation of novel, complex, human patient–specific models to deliver research breakthroughs.

scholarly journals Evaluation of Noxious Effect of Environmental Contaminants Using Zebrafish Model

2020 ◽  
Vol 2 (1) ◽  
pp. 37
Keyword(s):  
Cell Death ◽  
Developmental Stages ◽  
Morphological Changes ◽  
Cell Damage ◽  
Developmental Toxicity ◽  
Environmental Pollutant ◽  
Toxicity Study ◽  
Zebrafish Embryos ◽  
Zebrafish Model

Effects of the inorganic chemicals Calcium Fluoride (CaF2) and Hexaflurosilicilic acid (H2SiF2) have been studied due to its excessive usage in drinking water plants, glass manufacturing etc. Toxicity studies on Zebrafish embryos have been carried out for CaF2 and H2SiF2 during the embryonic developmental stages to observe the changes taken place during the growth, development. These changes can be observed in cell differentiation, larval movements, delay in hatching, and by the changes in behavior. Due to the ease with the transparency of zebrafish embryos, it can be observed and manipulated. In the field of early developmental studies, these zebrafish embryos have been vital because they have faster development by which the whole organs get developed in 3 days. Thus it plays a significant role in the discovery and analysis of changes in the developmental aspects of their teratology study. Toxicity study in Adults Zebrafish can be studied through the histology analysis where the cell damage and cell death due to fluorides and acid ions which may also lead to morphological changes due to this environmental pollutant. This toxicity study can be studied based on behavioral effects, LC50 determination, and immunohistochemistry of the brain to observe the developmental neurotoxicity. This study describes the effect of the inorganic chemicals is leading to developmental toxicity, cell deformities, and cell death with the high mortality rate in the In vivo Zebrafish model.

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