scholarly journals Gata3Hypomorphic Mutant Mice Rescued with a Yeast Artificial Chromosome Transgene Suffer a Glomerular Mesangial Cell Defect

2016 ◽  
Vol 36 (17) ◽  
pp. 2272-2281 ◽  
Author(s):  
Takashi Moriguchi ◽  
Lei Yu ◽  
Akihito Otsuki ◽  
Keiko Ainoya ◽  
Kim-Chew Lim ◽  
...  
Keyword(s):  
Kidney Development ◽  
Yeast Artificial Chromosome ◽  
Mesangial Cells ◽  
Critical Role ◽  
Artificial Chromosome ◽  
Mutant Mice ◽  
Renal Tubules ◽  
Glomerular Mesangial Cells ◽  
Renal Hypoplasia ◽  
Transgenic Mouse Line

GATA3 is a zinc finger transcription factor that plays a crucial role in embryonic kidney development, while its precise functions in the adult kidney remain largely unexplored. Here, we demonstrate that GATA3 is specifically expressed in glomerular mesangial cells and plays a critical role in the maintenance of renal glomerular function. Newly generatedGata3hypomorphic mutant mice exhibited neonatal lethality associated with severe renal hypoplasia. Normal kidney size was restored by breeding the hypomorphic mutant with a rescuing transgenic mouse line bearing a 662-kbGata3yeast artificial chromosome (YAC), and these animals (termed G3YR mice) survived to adulthood. However, most of the G3YR mice showed degenerative changes in glomerular mesangial cells, which deteriorated progressively during postnatal development. Consequently, the G3YR adult mice suffered severe renal failure. We found that the 662-kbGata3YAC transgene recapitulatedGata3expression in the renal tubules but failed to direct sufficient GATA3 activity to mesangial cells. Renal glomeruli of the G3YR mice had significantly reduced amounts of platelet-derived growth factor receptor (PDGFR), which is known to participate in the development and maintenance of glomerular mesangial cells. These results demonstrate a critical role for GATA3 in the maintenance of mesangial cells and its absolute requirement for prevention of glomerular disease.

scholarly journals Transgenic overexpression of GLUT1 in mouse glomeruli produces renal disease resembling diabetic glomerulosclerosis

2010 ◽  
Vol 299 (1) ◽  
pp. F99-F111 ◽  
Author(s):  
Youli Wang ◽  
Kathleen Heilig ◽  
Thomas Saunders ◽  
Andrew Minto ◽  
Dilip K. Deb ◽  
...  
Keyword(s):  
Diabetic Nephropathy ◽  
Transforming Growth Factor ◽  
Mesangial Cells ◽  
Glomerular Disease ◽  
Membrane Thickness ◽  
Renal Tubules ◽  
Glomerular Mesangial Cells ◽  
Diabetic Glomerulosclerosis ◽  
Basement Membrane Thickness ◽  
Intracellular Glucose

Previous work identified an important role for hyperglycemia in diabetic nephropathy (The Diabetes Control and Complications Trial Research Group. N Engl J Med 329: 977–986, 1993; UK Prospective Diabetes Study Group. Lancet 352: 837–853, 1998), and increased glomerular GLUT1 has been implicated. However, the roles of GLUT1 and intracellular glucose have not been determined. Here, we developed transgenic GLUT1-overexpressing mice (GT1S) to characterize the roles of GLUT1 and intracellular glucose in the development of glomerular disease without diabetes. GLUT1 was overexpressed in glomerular mesangial cells (MC) of C57BL6 mice, a line relatively resistant to diabetic nephropathy. Blood pressure, blood glucose, glomerular morphometry, matrix proteins, cell signaling, transcription factors, and selected growth factors were examined. Kidneys of GT1S mice overexpressed GLUT1 in glomerular MCs and small vessels, rather than renal tubules. GT1S mice were neither diabetic nor hypertensive. Glomerular GLUT1, glucose uptake, mean capillary diameter, and mean glomerular volume were all increased in the GT1S mice. Moderately severe glomerulosclerosis (GS) was established by 26 wk of age in GT1S mice, with increased glomerular type IV collagen and fibronectin. Modest increases in glomerular basement membrane thickness and albuminuria were detected with podocyte foot processes largely preserved, in the absence of podocyte GLUT1 overexpression. Activation of glomerular PKC, along with increased transforming growth factor-β1, VEGFR1, VEGFR2, and VEGF were all detected in glomeruli of GT1S mice, likely contributing to GS. The transcription factor NF-κB was also activated. Overexpression of glomerular GLUT1, mimicking the diabetic GLUT1 response, produced numerous features typical of diabetic glomerular disease, without diabetes or hypertension. This suggested GLUT1 may play an important role in the development of diabetic GS.

scholarly journals Endothelin signaling via guanine exchange factor C3G in renal glomerular mesangial cellsThis article is one of a selection of papers published in the two-part special issue entitled 20 Years of Endothelin Research.

2010 ◽  
Vol 88 (8) ◽  
pp. 808-816 ◽  
Author(s):  
Victoriya A. Rufanova ◽  
Anna Alexanian ◽  
Tammo Ostendorf ◽  
Dirk Bokemeyer ◽  
Simon Prosser ◽  
...  
Keyword(s):  
Intracellular Signaling ◽  
Kidney Development ◽  
Mesangial Cells ◽  
Biological Significance ◽  
Fiber Formation ◽  
Actin Dynamics ◽  
Glomerular Mesangial Cells ◽  
Nucleotide Exchange ◽  
Exchange Factor ◽  
Mesangioproliferative Glomerulonephritis

The guanine nucleotide exchange factor C3G is one of the mediators of endothelin-1 (ET-1) intracellular signaling cascades and is vital for kidney development and homeostasis. The aim of the current study was to analyze the specificity of ET-1-induced signaling via C3G in rat glomerular mesangial cells (GMC) and to investigate the biological significance of C3G during mesangioproliferative glomerulonephritis. In GMC, C3G expression was increased (1) in vivo after induction of the anti-Thy1 model of glomerulonephritis and (2) in cell culture experiments after fetal bovine serum incubation. To examine the consequences of C3G up-regulation, adenovirus-mediated gene transfer of C3G into cultured glomerular cells was done, and the GTP loading of the small G proteins Rap1 and R-Ras was analyzed. Overexpression of C3G in mesangial cells resulted in enhanced activation of Rap1, but failed to affect the GTP-bound status of R-Ras in ET-1-stimulated cells. C3G overexpression led to significant changes in GMC spreading and migration patterns in response to ET-1 stimulation and increased stress fiber formation, which was mimicked by Rap1A overexpression. Together, these findings suggest (1) the existence of regulatory mechanisms resulting in disease-related up-regulation of C3G in GMC and (2) that an increase in the C3G protein level may contribute to the resolution stage of mesangioproliferative glomerulonephritis by reducing GMC sensitivity to ET-1, modulating cellular motility, and actin dynamics.

scholarly journals Deficiency in Six2 during prenatal development is associated with reduced nephron number, chronic renal failure, and hypertension in Br/+ adult mice

2009 ◽  
Vol 296 (5) ◽  
pp. F1166-F1178 ◽  
Author(s):  
Ben Fogelgren ◽  
Shiming Yang ◽  
Ian C. Sharp ◽  
Odaro J. Huckstep ◽  
Wenbin Ma ◽  
...  
Keyword(s):  
Renal Failure ◽  
Chronic Renal Failure ◽  
Prenatal Development ◽  
Mutant Mice ◽  
Renal Tubules ◽  
Nephron Number ◽  
Renal Hypoplasia ◽  
Adult Mice ◽  
Homeobox Transcription Factor ◽  
And Function

The Br/+ mutant mouse displays decreased embryological expression of the homeobox transcription factor Six2, resulting in hertitable renal hypoplasia. The purpose of this study was to characterize the renal physiological consequences of embryonic haploinsuffiency of Six2 by analyzing renal morphology and function in the adult Br heterozygous mutant. Adult Br/+ kidneys weighed 50% less than those from wild-type mice and displayed glomerulopathy. Stereological analysis of renal glomeruli showed that Br/+ kidneys had an average of 88% fewer glomeruli than +/+ kidneys, whereas individual glomeruli in Br/+ mice maintained an average volume increase of 180% compared with normal nephrons. Immunostaining revealed increased levels of endothelin-1 (ET-1), endothelin receptors A (ETA) and B (ETB), and Na-K-ATPase were present in the dilated renal tubules of mutant mice. Physiological features of chronic renal failure (CRF) including elevated mean arterial pressure, increased plasma creatinine, and dilute urine excretion were measured in Br/+ mutant mice. Electron microscopy of the Br/+ glomeruli revealed pathological alterations such as hypercellularity, extracellular matrix accumulation, and a thick irregular glomerular basement membrane. These results indicate that adult Br/+ mice suffer from CRF associated with reduced nephron number and renal hypoplasia, as well as glomerulopathy. Defects are associated with embryological deficiencies of Six2, suggesting that proper levels of this protein during nephrogenesis are critical for normal glomerular development and adult renal function.

Effect of high glucose on gene expression in mesangial cells: upregulation of the thiol pathway is an adaptational response

2004 ◽  
Vol 17 (3) ◽  
pp. 271-282 ◽  
Author(s):  
Jolean Morrison ◽  
Kristen Knoll ◽  
Martin J. Hessner ◽  
Mingyu Liang
Keyword(s):  
Lipid Peroxidation ◽  
Mrna Expression ◽  
High Glucose ◽  
Molecular Mechanisms ◽  
Mesangial Cells ◽  
Critical Role ◽  
Glomerular Mesangial Cells ◽  
Human Mesangial Cells ◽  
Stage Renal Disease ◽  
End Stage

Pathological alterations in glomerular mesangial cells play a critical role in the development of diabetic nephropathy, the leading cause of end-stage renal disease. Molecular mechanisms mediating such alterations, however, remain to be fully understood. The present study first examined the effect of high glucose on the mRNA expression profile in rat mesangial cells using cDNA microarray. Based on variation-weighted criteria and with a false discovery rate of 4.3%, 459 of 17,664 cDNA elements examined were found to be upregulated and 151 downregulated by exposure to 25 mM d-glucose for 5 days. A large number of differentially expressed genes belonged to several functional categories, indicating high glucose had a profound effect on mesangial cell proliferation, protein synthesis, energy metabolism, and, somewhat unexpectedly, protein sorting and the cytoskeleton. Interestingly, several thiol antioxidative genes (glutathione peroxidase 1, peroxiredoxin 6, and thioredoxin 2) were found by microarray and confirmed by real-time PCR to be upregulated by high glucose. These changes suggested that the oxidative stress known to be induced in mesangial cells by high glucose might be buffered by upregulation of the thiol antioxidative pathway. Upregulation of thiol antioxidative genes also occurred in high-glucose-treated human mesangial cells and in glomeruli isolated from rats after 1 wk of streptozotocin-induced diabetes, but not in human proximal tubule cells. High glucose slightly increased lipid peroxidation and decreased the amount of reduced thiols in rat and human mesangial cells. Disruption of the thiol antioxidative pathway by two different thiol-oxidizing agents resulted in a three- to fivefold increase in high-glucose-induced lipid peroxidation. In summary, the present study provided a global view of the short-term effect of high glucose on mesangial cells at the level of mRNA expression and identified the upregulation of the thiol antioxidative pathway as an adaptational response of mesangial cells to high glucose.

Constitutive Activation of Smoothened in the Renal Collecting Ducts Leads to Renal Hypoplasia, Hydronephrosis, and Hydroureter

2017 ◽  
Vol 204 (1) ◽  
pp. 38-48 ◽  
Author(s):  
Deepak Prasad Gupta ◽  
Jae-Won Hwang ◽  
Eui-Sic Cho ◽  
Won Kim ◽  
Chang Ho Song ◽  
...  
Keyword(s):  
Kidney Development ◽  
Mutant Mice ◽  
Renal Hypoplasia ◽  
Shh Signaling ◽  
Constitutive Activation ◽  
Downstream Target ◽  
Collecting Ducts ◽  
Downstream Target Gene ◽  
Patched 1 ◽  
First Time

Sonic Hedgehog (Shh) signaling plays a major role in and is essential for regulation, patterning, and proliferation during renal development. Smoothened (Smo) plays a pivot role in transducing the Shh-glioma-associated oncogene Kruppel family member. However, the cellular and molecular mechanism underlying the role of sustained Smo activation in postnatal kidney development is still not clearly understood. Using a conditional knockin mouse model that expresses a constitutively activated form of Smo (SmoM2) upon Homeobox-B7-mediated recombination (Hoxb7-Cre), the effects of Shh signaling were determined in postnatal kidney development. SmoM2;Hoxb7-Cre mutant mice showed growth retardation with a reduction of body weight. Constitutive activation of Smo in the renal collecting ducts caused renal hypoplasia, hydronephrosis, and hydroureter. The parenchymal area and glomerular numbers were reduced, but the glomerular density was increased in SmoM2;Hoxb7-Cre mutant mice. The expression of Patched 1, the receptor of Shh and a downstream target gene of the Shh signaling pathway, was highly restricted and it was upregulated in the inner medullary collecting ducts of the kidney. The proliferative cells in the mesenchyme and collecting ducts were decreased in SmoM2;Hoxb7-Cre mutant mice. This study showed for the first time that sustained Smo inhibits postnatal kidney development by suppressing the proliferation of the mesenchyme and medullary collecting ducts in mice.

Rescue of the lethal scl−/− phenotype by the human SCL locus

Blood ◽  
2002 ◽  
Vol 99 (11) ◽  
pp. 3931-3938 ◽  
Author(s):  
Angus M. Sinclair ◽  
Anthony J. Bench ◽  
Adrian J. C. Bloor ◽  
Juan Li ◽  
Berthold Göttgens ◽  
...  
Keyword(s):  
Yeast Artificial Chromosome ◽  
Fetal Liver ◽  
Critical Role ◽  
Artificial Chromosome ◽  
Regulatory Elements ◽  
Yolk Sac ◽  
Loss Of Function ◽  
Hematopoietic Stem ◽  
Helix Loop Helix

The stem cell leukemia (SCL) gene encodes a basic helix-loop-helix transcription factor with a critical role in the development of both blood and endothelium. Loss-of-function studies have shown that SCL is essential for the formation of hematopoietic stem cells, for subsequent erythroid development and for yolk sac angiogenesis. SCL exhibits a highly conserved pattern of expression from mammals to teleost fish. Several murine SCLenhancers have been identified, each of which directs reporter gene expression in vivo to a subdomain of the normal SCL expression pattern. However, regulatory elements necessary for SCL expression in erythroid cells remain to be identified and the size of the chromosomal domain needed to support appropriate SCL transcription is unknown. Here we demonstrate that a 130-kilobase (kb) yeast artificial chromosome (YAC) containing the human SCL locus completely rescued the embryonic lethal phenotype ofscl−/− mice. Rescued YAC+scl−/− mice were born in appropriate Mendelian ratios, were healthy and fertile, and exhibited no detectable abnormality of yolk sac, fetal liver, or adult hematopoiesis. The human SCL protein can therefore substitute for its murine homologue. In addition, our results demonstrate that the human SCL YAC contains the chromosomal domain necessary to direct expression to the erythroid lineage and to all other tissues in which SCL performs a nonredundant essential function.

scholarly journals Berberine Interferes With the Abnormal Glomerular Mesangial Cell C Ycle Re-Distribution to Alleviate Diabetic Nephropathy via PI3K/AKT/AS160/GLUT4 Signaling Pathway

2021 ◽  
Author(s):  
Ximei Guan ◽  
Weijian Ni ◽  
Jing Zeng ◽  
Hong Zhou ◽  
Linqin Tang
Keyword(s):  
Cell Cycle ◽  
Diabetic Nephropathy ◽  
Glucose Uptake ◽  
Signaling Pathway ◽  
High Glucose ◽  
Mesangial Cells ◽  
Critical Role ◽  
G1 Phase ◽  
Glomerular Mesangial Cells ◽  
Time Flow

Abstract Background: Berberine plays a critical role of the glomerular mesangial cells (GMCs) abnormal proliferation during diabetic nephropathy (DN). This study aims to explore the intervention effect of BBR on DN mice and investigate the potential mechanism targeting abnormal GMCs proliferation. Methods: Streptozotocin-induced mice were used to determine the effect of BBR on the renal injury. In vitro, GMCs are cultured in high glucose (30 mmol/L). EdU and MTT assay are used for screening the optimum BBR concentration and intervention time. Flow cytometry is applied to analyze the cell cycle re-distribution. Western blot and RT-qPCR are devoted to studying the relative expression of molecules in PI3K/AKT/AS160/GLUT4 signaling pathway. Additionally, 2-NBDG assay is selected for measuring the glucose uptake of GMCs. Results: HE and PAS staining revealed that the notable mesangial matrix expansion, glomerular hypertrophy and glycogen deposition in diabetic kidney can be alleviated after BBR treatment. Moreover, BBR significantly reduced the positive expression of GLUT4 in tubulointerstitium and glomerular region. EdU shows that high glucose induces the abnormal proliferation of GMCs, which becomes more apparently as time goes on (20h→28h). Meantime, BBR (60 and 90 μmol/L) can not only increase the proportion of G1 phase, but also reduce the proportion of S phase. After 24 h, the same sort of phenomenon has cropped up in BBR (30 μmol/L) group. BBR (60 μmol/L)) significantly degraded the levels of PI3K-p85, p-AKT, p-AS160, and the membrane-GLUT4, while indistinguishable changes of their total protein expressions. Additionally, BBR prominently reduced the glucose uptake and retard the cell cycle of GMCs to stay at G1 phase. Conclusions: The rearrangement effect of BBR on cell cycle is related with PI3K/AKT signaling pathway and GLUT4 trafficking. The key findings of our study collectively indicate that the treatment of GMCs proliferation can be a novel therapeutic strategy in DN.

Platelet-activating factor and the kidney

1986 ◽  
Vol 251 (1) ◽  
pp. F1-F11 ◽  
Author(s):  
D. Schlondorff ◽  
R. Neuwirth
Keyword(s):  
De Novo ◽  
Mesangial Cells ◽  
Biological Activities ◽  
Platelet Activating Factor ◽  
Calcium Ionophore ◽  
Initial Step ◽  
Renal Physiology ◽  
Dependent Manner ◽  
Glomerular Mesangial Cells ◽  
Isolated Kidney

Platelet-activating factor (PAF) represents a group of phospholipids with the basic structure of 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine. A number of different cells are capable of producing PAF in response to various stimuli. The initial step of PAF formation is activation of phospholipase A2 in a calcium-dependent manner, yielding lyso-PAF. During this step arachidonic acid is also released and can be converted to its respective cyclooxygenase and lipoxygenase products. The lyso-PAF generated is then acetylated in position 2 of the glycerol backbone by a coenzyme A (CoA)-dependent acetyltransferase. An additional pathway may exist whereby PAF is generated de novo from 1-alkyl-2-acetyl-sn-glycerol by phosphocholine transferase. PAF inactivation in cells and blood is by specific acetylhydrolases. PAF exhibits a variety of biological activities including platelet and leukocyte aggregation and activation, increased vascular permeability, respiratory distress, decreased cardiac output, and hypotension. In the kidney PAF can produce decreases in blood flow, glomerular filtration, and fluid and electrolyte excretion. Intrarenal artery injection of PAF may also result in glomerular accumulation of platelets and leukocytes and mild proteinuria. PAF increases prostaglandin formation in the isolated kidney and in cultured glomerular mesangial cells. PAF also causes contraction of mesangial cells. Upon stimulation with calcium ionophore the isolated kidney, isolated glomeruli and medullary cells, and cultured mesangial cells are capable of producing PAF. The potential role for PAF in renal physiology and pathophysiology requires further investigation that may be complicated by 1) the multiple interactions of PAF, prostaglandins, and leukotrienes and 2) the autocoid nature of PAF, which may restrict its action to its site of generation.

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