Isolation of Glomeruli from Mammalian Kidneys by Graded Sieving

Rp, Misra

doi:10.1093/ajcp/58.2.135

Cited by 177 publications

(88 citation statements)

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“…Neonatal rat CMCs were obtained from 2-to 3-day-old rats and cultured as described (38). Glomeruli were isolated by differential sieving as published (39). CMC and glomeruli were stimulated at 37°C for 20 min with different concentrations of ANP or CNP in the presence of 0.5 mM 3-isobutyl-1-methylxanthine (Sigma).…”

Section: Methodsmentioning

confidence: 99%

Cardiac hypertrophy in transgenic rats expressing a dominant-negative mutant of the natriuretic peptide receptor B

Langenickel

Buttgereit

Pagel-Langenickel

et al. 2006

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

115

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Natriuretic peptides (NP) mediate their effects by activating membrane-bound guanylyl cyclase-coupled receptors A (NPR-A) or B (NPR-B). Whereas the pathophysiological role of NPR-A has been widely studied, only limited knowledge on the cardiovascular function of NPR-B is available. In vitro studies suggest antiproliferative and antihypertrophic actions of the NPR-B ligand C-type NP (CNP). Because of the lack of a specific pharmacological inhibitor, these effects could not clearly be attributed to impaired NPR-B signaling. Recently, gene deletion revealed a predominant role of NPR-B in endochondral ossification and development of female reproductive organs. However, morphological abnormalities and premature death of NPR-B-deficient mice preclude detailed cardiovascular phenotyping. In the present study, a dominant-negative mutant (NPR-B⌬KC) was used to characterize CNP-dependent NPR-B signaling in vitro and in transgenic rats. Here we demonstrate that reduced CNP-but not atrial NP-dependent cGMP response attenuates antihypertrophic potency of CNP in vitro. In transgenic rats, NPR-B⌬KC expression selectively reduced NPR-B but not NPR-A signaling. NPR-B⌬KC transgenic rats display progressive, blood pressure-independent cardiac hypertrophy and elevated heart rate. The hypertrophic phenotype is further enhanced in chronic volume overload-induced congestive heart failure. Thus, this study provides evidence linking NPR-B signaling to the control of cardiac growth.C-type natriuretic peptide ͉ knockdown

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Section: Methodsmentioning

confidence: 99%

Cardiac hypertrophy in transgenic rats expressing a dominant-negative mutant of the natriuretic peptide receptor B

Langenickel

Buttgereit

Pagel-Langenickel

et al. 2006

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

115

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“…Therefore, we collected the glomeruli from the rat kidneys using a standard sieving method. 26 To clarify the mechanisms underlying NTS-induced GN and the favorable effects of the pCAGGS-vIL-10 transfer, we performed quantitative real-time PCR analysis for the mRNA expression levels of CD4, CD8, IFN-g, TNF-a, perforin (lytic molecules produced by CD8 + T cells), 27 and monocyte chemotactic protein (MCP)-1 in the glomeruli of normal rat kidneys before NTS injection and of kidneys from pCAGGS and pCAGGS-vIL-10 rats on days 4 and 7 after NTS injection (n¼5 in each group) ( Figure 7). The mean values with their s.d.…”

Section: Effect Of Pcaggs-vil-10 Transfer On Glomerular Lesionsmentioning

confidence: 99%

“…The 20 kidneys were removed and the cortices were dissected into small pieces. The cortices were then sieved according to a standard sieving method, 26 using a series of sieves of 250, 150, and 75 mm (Sansyo, Tokyo, Japan) to isolate the vIL-10 gene transfer in a glomerulonephritis model N Higuchi et al glomeruli, and the GBM was subsequently isolated using the method of Meezan et al 44 Production of NTS containing rabbit polyclonal anti-rat GBM antibodies…”

Section: Preparation Of Rat Gbmmentioning

confidence: 99%

Hydrodynamics-based delivery of the viral interleukin-10 gene suppresses experimental crescentic glomerulonephritis in Wistar–Kyoto rats

et al. 2003

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Gene therapy is expected to revolutionize the treatment of kidney diseases. Viral interleukin (vIL)-10 has a variety of immunomodulatory properties. We examined the applicability of vIL-10 gene transfer to the treatment of rats with crescentic glomerulonephritis, a T helper 1 (Th 1) predominant disease. To produce the disease, Wistar-Kyoto rats were injected with a rabbit polyclonal anti-rat glomerular basement membrane antibody. After 3 h, a large volume of plasmid DNA expressing vIL-10 (pCAGGS-vIL-10) solution was rapidly injected into the tail vein. pCAGGS solution was similarly injected into control rats (pCAGGS rats). We confirmed the presence of vector-derived vIL-10 mainly in the liver and observed high serum vIL-10 levels in pCAGGSvIL-10-injected rats. Compared with the pCAGGS rats, the pCAGGS-vIL-10 rats showed significant therapeutic effects: reduced frequency of crescent formation, decrease in the number of total cells, macrophages, and CD4 + T cells in the glomeruli, decrease in urine protein, and attenuation of kidney dysfunction. Using quantitative real-time polymerase chain reaction, we also observed that this model was Th1-predominant in the glomeruli and that the ratio of the transcripts of CD4, interferon-g, tumor necrosis factor-a, and monocyte chemotactic protein-1 to the transcripts of glucose-6-phosphate dehydrogenase in the glomeruli were all significantly lower in the pCAGGS-vIL-10 rats than in the pCAGGS rats. These results demonstrate that pCAGGS-vIL-10 gene transfer by hydrodynamics-based transfection suppresses crescentic glomerulonephritis.

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“…Glomeruli were isolated from fresh bovine or human kidneys by a sieving method (15) and stored frozen. After thawing, the suspension was centrifuged at 2,000 g for 10 rain, and the pellet was suspended in 100 vol distilled water containing protease inhibitors (0.1 M 6-aminohexanoic acid, 0.01 M EDTA, and 0.005 M benzamidine-HCl), stirred for 3 h, and centrifuged at 17,700 g for 10 min.…”

mentioning

confidence: 99%

Sera from patients with poststreptococcal glomerulonephritis contain antibodies to glomerular heparan sulfate proteoglycan.

Fillit¹,

Damle²,

Gregory³

et al. 1985

The Journal of Experimental Medicine

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Proteoglycans (PG) ~ are important tissue components,both in the extracellular matrix (1) and as cell membrane-associated components (2). They are composed of protein and glycosaminoglycans (GAG), containing glucosamine or galactosamine and (generally) uronic acid. About fifty years ago, Goebel (3) demonstrated the antigenicity of protein-bound uronic acid and, further, showed that rabbit antisera could distinguish protein-bound glucuronic from galacturonic acid (4). More recently, the presence of immunity to heparan sulfate and chondroitin sulfate PG has been demonstrated in animals (5-7). Both protein core and GAG determinants are antigenic. Studies of systemic lupus erythematosus (SLE) sera (8) suggest that antibodies to DNA may cross-react with hyaluronate and chondroitin sulfate on a nonspecific charge basis because of the repeating anionic sites that DNA and GAG have in common.Previous studies (9-13) suggest that autoimmunity to a number of kidney antigens may exist in acute and chronic giomerulonephritis (GN). Our own work (13,14) suggests that the carbohydrate antigens of the giomerular basement membrane (GBM) may be antigenic. Since one of the major carbohydratecontaining antigens of the GBM is heparan sulfate PG (1), we studied the sera of patients with acute poststreptococcal glomerulonephritis (PSGN) for the presence of antibodies to glomerular PG. These studies demonstrate the presence, in PSGN sera, of antibodies to glomerular PG. Heparan sulfate is the primary target of immune reactivity. Mammalian hyaluronate and streptococcal hyaluronate inhibited the observed reactivity, suggesting cross-reactivity in the immune response to heparan sulfate, mammalian, and streptococcal hyaluronate. A second site of immunoreactivity contains N-acetylgalactosamine, possibly representing autoimmunity to chondroitin or dermatan sulfate PG.

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Isolation of Glomeruli from Mammalian Kidneys by Graded Sieving

Cited by 177 publications

References 0 publications

Cardiac hypertrophy in transgenic rats expressing a dominant-negative mutant of the natriuretic peptide receptor B

Cardiac hypertrophy in transgenic rats expressing a dominant-negative mutant of the natriuretic peptide receptor B

Hydrodynamics-based delivery of the viral interleukin-10 gene suppresses experimental crescentic glomerulonephritis in Wistar–Kyoto rats

Sera from patients with poststreptococcal glomerulonephritis contain antibodies to glomerular heparan sulfate proteoglycan.

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