Podocyte‐specific expression of cre recombinase in transgenic mice

Moeller, Marcus J.; Sanden, Silja K.; Soofi, Abdulsalam; Wiggins, Roger C.; Holzman, Lawrence B.

doi:10.1002/gene.10164

Cited by 282 publications

(325 citation statements)

References 13 publications

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“…To study the effects of proximal tubule-specific overexpression on the phenotype in the L/L Akita mice, we used the Ggt1 promoterdriven Cre transgene (Ggt1-cre/ERT2; European Mouse Mutant Archive) (25), which is induced by tamoxifen injection (50 mg/kg/day IP in sesame oil for 5 d) at age 4 wk. To study the effects of podocyte-specific overexpression on the phenotype in the L/L Akita mice, we used the podocin (Nphs2) promoterdriven Cre transgene (Nphs2-cre; The Jackson Laboratory) (26). All mice were kept under the husbandry conditions in conformance with guidelines of University of North Carolina Institutional Animal Care and Use Committee.…”

Section: Methodsmentioning

confidence: 99%

Low TGFβ1 expression prevents and high expression exacerbates diabetic nephropathy in mice

Hathaway

Gasim

Grant

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Nephropathy develops in many but not all patients with longstanding type 1 diabetes. Substantial efforts to identify genotypic differences explaining this differential susceptibility have been made, with limited success. Here, we show that the expression of the transforming growth factor β1 gene (Tgfb1) affects the development of diabetic nephropathy in mice. To do this we genetically varied Tgfb1 expression in five steps, 10%, 60%, 100%, 150%, and 300% of normal, in mice with type 1 diabetes caused by the Akita mutation in the insulin gene (Ins2 Akita ). Although plasma glucose levels were not affected by Tgfb1 genotype, many features of diabetic nephropathy (mesangial expansion, elevated plasma creatinine and urea, decreased creatinine clearance and albuminuria) were progressively ameliorated as Tgfb1 expression decreased and were progressively exacerbated when expression was increased. The diabetic 10% hypomorphs had comparable creatinine clearance and albumin excretion to wild-type mice and no harmful changes in renal morphology. The diabetic 300% hypermorphs had ∼1/3 the creatinine clearance of wild-type mice, >20× their albumin excretion, ∼3× thicker glomerular basement membranes and severe podocyte effacement, matching human diabetic nephropathy. Switching Tgfb1 expression from low to high in the tubules of the hypomorphs increased their albumin excretion more than 10-fold but creatinine clearance remained high. Switching Tgfb1 expression from low to high in the podocytes markedly decreased creatinine clearance, but minimally increased albumin excretion. Decreasing expression of Tgfb1 could be a promising option for preventing loss of renal function in diabetes.aldosterone | glomerular filtration rate | glomerulosclerosis | megalin | nephrin D iabetes is the number one cause of end-stage renal disease in the United States and many other developed countries. However, despite having similar levels of blood glucose only 20-40% of all diabetic patients develop diabetic nephropathy. In diabetic nephropathy, increased expression of transforming growth factor β1 (TGFβ1) has been demonstrated to promote accumulation of extracellular matrix components (1), apoptosis (2), dedifferentiation of podocytes (3), and epithelial-mesenchymal transition of proximal tubules (4), all of which are thought to facilitate a decline in nephron number and renal function.Tgfb1-null mice on a mixed genetic background show severe multiorgan inflammation with massive infiltration of lymphocytes and macrophages that culminates in death by 3-4 wk of age (5, 6). Their death effectively prevents determining whether absence of TGFβ1 influences the development of nephropathy. To overcome this problem and also to allow the study of the effects of above-normal TGFβ1, we have generated mice with five genetically graded levels of TGFβ1, and have made them diabetic with the Ins2 Akita mutation, which causes pancreatic beta-cell dysfunction and type 1 diabetes.Here we show that the features characteristic of diabetic nephropathy are progressively ...

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Section: Methodsmentioning

confidence: 99%

Low TGFβ1 expression prevents and high expression exacerbates diabetic nephropathy in mice

Hathaway

Gasim

Grant

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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“…The transcription start site of the nephrin mRNA is reported to be either at nucleotide Ϫ415 or Ϫ381, as counted from the nephrin translation initiation ATG codon in the cDNA (30,32). To construct pEGFP-N5Ј (391bp) -enhanced green fluorescent protein (GFP), a 391-nucleotide fragment corresponding to the mouse nephrin 5Ј-flanking region (nucleotides Ϫ391 to 1) was produced using PCR.…”

Section: Methodsmentioning

confidence: 99%

Role of the Endoplasmic Reticulum Unfolded Protein Response in Glomerular Epithelial Cell Injury

Cybulsky¹,

Takano²,

Papillon

et al. 2005

Journal of Biological Chemistry

104

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C5b-9-induced glomerular epithelial cell (GEC) injury in vivo (in passive Heymann nephritis) and in culture is associated with damage to the endoplasmic reticulum (ER) and increased expression of ER stress proteins. Induction of ER stress proteins is enhanced via cytosolic phospholipase A 2 (cPLA 2 ) and limits complement-dependent cytotoxicity. The present study addresses another aspect of the ER unfolded protein response, i.e. activation of protein kinase R-like ER kinase (PERK or pancreatic ER kinase), which phosphorylates eukaryotic translation initiation factor 2-␣ (eIF2␣), thereby generally suppressing translation and decreasing the protein load on a damaged ER. Phosphorylation of eIF2␣ was enhanced significantly in glomeruli of proteinuric rats with passive Heymann nephritis, compared with control. In cultured GECs, complement induced phosphorylation of eIF2␣ and reduced protein synthesis, and complement-stimulated phosphorylation of eIF2␣ was enhanced by overexpression of cPLA 2 . Ischemia-reperfusion in vitro (deoxyglucose plus antimycin A followed by glucose re-exposure) also stimulated eIF2␣ phosphorylation and reduced protein synthesis. Complement and ischemia-reperfusion induced phosphorylation of PERK (which correlates with activation), and fibroblasts from PERK knock-out mice were more susceptible to complement-and ischemia-reperfusion-mediated cytotoxicity, as compared with wild type fibroblasts. The GEC protein, nephrin, plays a key role in maintaining glomerular permselectivity. In contrast to a general reduction in protein synthesis, translation regulated by the 5-end of mouse nephrin mRNA during ER stress was paradoxically maintained, probably due to the presence of short open reading frames in this mRNA segment. Thus, phosphorylation of eIF2␣ and consequent general reduction in protein synthesis may be a novel mechanism for limiting complement-or ischemiareperfusion-dependent GEC injury.

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“…Heterozygous ROSA26 mice (C57BL/6 background) were obtained from The Jackson Laboratory (Bar Harbor, ME) and crossed with 2.5P-Cre mice (F2N3, 87.5% C57BL/6J/12.5% SJL/J background) (12). Animals were maintained under specific pathogen-free conditions.…”

Section: Animalsmentioning

confidence: 99%

“…Transgenic mice were identified by using a PCR strategy with DNA recovered from tail biopsies, as described previously (12,13). The RO-SA26 transgene was identified with the primers LacZ.fwd (TTCACTG-GCCGTCGTTTTACAACGTCGTGA) and LacZ.rev (ATGTGAGC-GAGTAACAACCCGTCGGATTCT).…”

Section: Identification Of Transgenic Micementioning

confidence: 99%

“…ROSA26 reporter mice were bred to mice of the 2.5P-Cretransgenic mouse line, which mediates Cre recombination driven by a 2.5-kb human NPHS2 (podocin) promoter fragment exclusively in podocytes (12). Cre excision of the floxed neo cassette allowed expression of ␤-galactosidase exclusively in podocytes of doubly transgenic mice (Figure 1).…”

Section: Somatic Cre Recombination As a Tool To Genetically Tag Podocmentioning

confidence: 99%

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Podocytes Populate Cellular Crescents in a Murine Model of Inflammatory Glomerulonephritis

Moeller

Soofi²,

Hartmann

et al. 2004

Journal of the American Society of Nephrology

168

142

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Abstract. Cellular crescents are a defining histologic finding in many forms of inflammatory glomerulonephritis. Despite numerous studies, the origin of glomerular crescents remains unresolved. A genetic cell lineage-mapping study with a novel transgenic mouse model was performed to investigate whether visceral glomerular epithelial cells, termed podocytes, are precursors of cells that populate cellular crescents. The podocytespecific 2.5P-Cre mouse line was crossed with the ROSA26 reporter line, resulting in irreversible constitutive expression of ␤-galactosidase in doubly transgenic 2.5P-Cre/ROSA26 mice. In these mice, crescentic glomerulonephritis was induced with a previously described rabbit anti-glomerular basement membrane antiserum nephritis approach. Interestingly, ␤-galactosidase-positive cells derived from podocytes adhered to the parietal basement membrane and populated glomerular crescents during the early phases of cellular crescent formation, accounting for at least one-fourth of the total cell mass. In cellular crescents, the proliferation marker Ki-67 was expressed in ␤-galactosidase-positive and ␤-galactosidase-negative cells, indicating that both cell types contributed to the formation of cellular crescents through proliferation in situ. Podocyte-specific antigens, including WT-1, synaptopodin, nephrin, and podocin, were not expressed by any cells in glomerular crescents, suggesting that podocytes underwent profound phenotypic changes in this nephritis model.

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Podocyte‐specific expression of cre recombinase in transgenic mice

Cited by 282 publications

References 13 publications

Low TGFβ1 expression prevents and high expression exacerbates diabetic nephropathy in mice

Low TGFβ1 expression prevents and high expression exacerbates diabetic nephropathy in mice

Role of the Endoplasmic Reticulum Unfolded Protein Response in Glomerular Epithelial Cell Injury

Podocytes Populate Cellular Crescents in a Murine Model of Inflammatory Glomerulonephritis

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