Teratogenic Effect of Puromycin Aminonucleoside on Rat Kidney

Atriopeptin is synthesized in mammalian atria as a 126-amino acid (14 kDa) prohormone, but it is secreted and circulates as a 28-amino acid (2.5 kDa) peptide. We have demonstrated the synthesis and secretion of an atriopeptin-like peptide in neonatal and adult rat kidney cell cultures. In this study, we evaluated the site of renal synthesis of this protein and its expression in normal rats and rats made nephrotic with puromycin aminonucleoside. The major form of atriopeptin in normal kidneys comigrated with an apparent molecular mass of 2.5 kDa assessed by gel filtration chromatography. However, the major form of this atriopeptin-like protein in nephrotic kidneys was determined to have an apparent molecular mass similar to the heart prohormone. No atriopeptin prohormone was detected in the plasma of nephrotic rats. Localization of this renal atriopeptin-like protein was accomplished by immunocytochemistry of rat kidney frozen sections. Using an antibody generated against either the COOH-terminal or NH3-terminal region of the cardiac atriopeptin prohormone, we detected specific immunostaining in the distal cortical nephron of the nephrotic kidney. This is the first report of the anatomic localization of a renal atriopeptin-like protein and its upregulation in nephrosis.

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Puromycin aminonucleoside nephrosis results in a marked increase in fractional clearance of albumin

AJP Renal Physiology ◽

10.1152/ajprenal.1999.277.1.f139 ◽

1999 ◽

Vol 277 (1) ◽

pp. F139-F145 ◽

Cited By ~ 10

Author(s):

Tanya M. Osicka ◽

Alexandra R. Hankin ◽

Wayne D. Comper

Keyword(s):

Rat Kidney ◽

Tubular Epithelium ◽

Size Selectivity ◽

Puromycin Aminonucleoside ◽

Glomerular Capillary Wall ◽

Perfused Rat Kidney ◽

Fractional Clearance ◽

Aminonucleoside Nephrosis ◽

Puromycin Aminonucleoside Nephrosis

Puromycin aminonucleoside nephrosis (PAN) results in a marked increase in the fractional clearance of albumin. The increase in the fractional clearance of [3H]albumin to ∼0.045, as measured both in vivo and in the isolated perfused rat kidney (IPK) with PAN, occurs without an accompanying equivalent increase in glomerular capillary wall size selectivity as previously measured with dextrans. This is very similar to the marked increase in albuminuria seen with kidneys treated with inhibitors of endocytosis by the tubular epithelium, particularly lysine (T. M. Osicka, L. M. Pratt, and W. D. Comper. Nephrology 2: 199–212, 1996). The similarity is further established that, like in the presence of lysine, [3H]albumin excreted in urine from rats with PAN is essentially intact whereas, in both in vivo and IPK control experiments, excreted [3H]albumin is heavily degraded. The same observations have also been made for3H-labeled anionic horseradish peroxidase. These observations suggest that the significant albuminuria that occurs in PAN is primarily post-glomerular basement membrane in origin.

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Taurine Supplementation Alleviates Puromycin Aminonucleoside Damage by Modulating Endoplasmic Reticulum Stress and Mitochondrial-Related Apoptosis in Rat Kidney

Nutrients ◽

10.3390/nu10060689 ◽

2018 ◽

Vol 10 (6) ◽

pp. 689 ◽

Cited By ~ 8

Author(s):

Alessandra Stacchiotti ◽

Gaia Favero ◽

Antonio Lavazza ◽

Maria Monsalve ◽

Luigi Rodella ◽

...

Keyword(s):

Endoplasmic Reticulum ◽

Endoplasmic Reticulum Stress ◽

Rat Kidney ◽

Taurine Supplementation ◽

Puromycin Aminonucleoside

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Renal neuraminidase. Characterization in normal rat kidney and measurement in experimentally induced nephrotic syndrome

Biochemical Journal ◽

10.1042/bj2390705 ◽

1986 ◽

Vol 239 (3) ◽

pp. 705-710 ◽

Cited By ~ 16

Author(s):

W H Baricos ◽

S Cortez-Schwartz ◽

S V Shah

Keyword(s):

Nephrotic Syndrome ◽

Specific Activity ◽

Rat Kidney ◽

Puromycin Aminonucleoside ◽

Glomerular Diseases ◽

Neuraminidase Activity ◽

Acetylneuraminic Acid ◽

Normal Rat ◽

High Concentrations ◽

Triton X

Several lines of evidence suggest that increased neuraminidase activity may be responsible for the loss of glomerular N-acetylneuraminic acid (AcNeu) observed in various glomerular diseases. However, virtually no information is available on the activity of neuraminidase in glomeruli or the potential role of this enzyme in glomerular pathophysiology. Utilizing 2′-(4-methylumbelliferyl)-alpha-D-N-acetylneuraminic acid (4MU-AcNeu) as substrate, we defined optimal assay conditions and characterized neuraminidase activity in glomeruli and, for comparison, in other renal fractions and liver. Neuraminidase activity in glomeruli, cortex and tubules was maximal at pH 4.4. The Km for 4MU-AcNeu was estimated to be 195 microM for glomeruli and 226 microM for cortex. Glomerular neuraminidase was inhibited by AcNeu (90% at 25 mM) and high concentrations of Triton X-100 (26% at 0.5%), but unaffected by CaCl2, EDTA or N-ethylmaleimide (each 1 mM). Neuraminidase activity (nmol/h per mg of protein; mean +/- S.E.M.) in normal rat kidney was: cortex, 14.47 +/- 0.76; medulla, 7.85 +/- 0.64; papilla, 2.64 +/- 0.11; tubules, 13.79 +/- 0.70; glomeruli, 5.57 +/- 0.28. In comparison, neuraminidase activity in rat liver was 2.58 +/- 0.14. Puromycin aminonucleoside (PAN)-induced nephrotic syndrome is a model of glomerular disease in which the loss of glomerular AcNeu is well documented. In two separate studies, we observed no change in the specific activity of neuraminidase in either glomeruli or cortex isolated from rats treated with PAN (15 mg/100 g, intraperitoneally) and killed at either the onset or the peak of proteinuria. Results were similar whether neuraminidase activity was expressed per mg of protein or per microgram of DNA.

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Rat glomerular epithelial cells in culture express characteristics of parietal, not visceral, epithelium.

Journal of the American Society of Nephrology ◽

10.1681/asn.v361279 ◽

1992 ◽

Vol 3 (6) ◽

pp. 1279-1287

Author(s):

T Weinstein ◽

R Cameron ◽

A Katz ◽

M Silverman

Keyword(s):

Epithelial Cells ◽

Intermediate Filaments ◽

Rat Kidney ◽

Parietal Cells ◽

Immunoperoxidase Staining ◽

Puromycin Aminonucleoside ◽

Coated Pits ◽

Glomerular Epithelial Cells

Glomerular epithelial cells (GEC) in culture are derived from intact isolated glomeruli. Although there is general agreement about distinguishing GEC from mesangial and endothelial cells, there is still uncertainty regarding the visceral versus parietal origin of cultured GEC. If these cells are to provide a useful model system, it is necessary to establish well-defined cell populations. The purpose of this study was to evaluate this important issue by comparing the characteristics of cultured GEC with glomerular epithelium from rat kidney sections. By electron microscopy, GEC were polygonal, with cilia and desmosomes between cells, similar to parietal cells in situ. Because intermediate filaments are specifically expressed in differentiated cells in the kidney, the expression of intermediate filaments in cultured GEC were compared with those of intact glomeruli. Cultured GEC are positive for cytokeratin and negative for vimentin and desmin, identical to parietal cells in situ. In contrast, podocytes are positive for vimentin and desmin and negative for cytokeratin. In vivo, podocytes express gp330 and puromycin-aminonucleoside (PAN) susceptibility, which are used as markers for cultured GEC. Immunoperoxidase staining of rat kidney sections with monoclonal anti-gp330 demonstrated gp330 localization to the cell surface and coated pits of the parietal cells, similar to its localization in podocytes. The presence of gp330 in cultured GEC was confirmed by immunoblot. PAN administration to rats induced vacuolization and detachment from the basement membrane in the parietal cells of Bowman's capsule, similar to the cytotoxicity observed in podocytes.(ABSTRACT TRUNCATED AT 250 WORDS)

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The effect of puromycin aminonucleoside on the incorporation of labeled precursors into rat-kidney RNA

Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis ◽

10.1016/0005-2787(66)90370-4 ◽

1966 ◽

Vol 129 (2) ◽

pp. 288-293 ◽

Cited By ~ 3

Author(s):

N. Dickie ◽

L.F. Norton ◽

R.F. Derr ◽

C.S. Alexander ◽

H.T. Nagasawa

Keyword(s):

Rat Kidney ◽

Puromycin Aminonucleoside

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Increased cathepsin D-like activity in cortex, tubules, and glomeruli isolated from rats with experimental nephrotic syndrome

Biochemical Journal ◽

10.1042/bj2230393 ◽

1984 ◽

Vol 223 (2) ◽

pp. 393-399 ◽

Cited By ~ 23

Author(s):

W H Baricos ◽

S V Shah

Keyword(s):

Nephrotic Syndrome ◽

Cathepsin D ◽

Rat Kidney ◽

Urinary Protein Excretion ◽

Urinary Protein ◽

Rat Tissues ◽

Puromycin Aminonucleoside ◽

Protein Excretion ◽

Normal Rat ◽

Functional Areas

We have examined the activity and distribution of cathepsin D (EC 3.4.23.5), a major renal lysosomal endoproteinase, in the various anatomical and functional areas of normal rat kidney. Cathepsin D-like activities (delta A280/h per mg of protein) in normal rat tissues were: cortex, 0.78 +/- 0.05, n = 37; medulla, 0.62 +/- 0.03, n = 12; papilla, 0.63 +/- 0.04, n = 12; tubules, 0.74 +/- 0.04, n = 28; glomeruli, 0.59 +/- 0.03, n = 28; and liver, 0.41 +/- 0.02, n = 28. Enzyme activity was maximal at pH 3.0-3.5 and inhibited more than 90% by pepstatin (6.7 micrograms/ml), suggesting that the enzyme is cathepsin D. In subsequent experiments we measured cathepsin D-like activity in cortex, tubules and glomeruli isolated from rats with puromycin aminonucleoside (PAN)-induced nephrotic syndrome. Treated animals (15 mg of PAN/100g body wt., intraperitoneally) developed proteinuria beginning 4 days after injection and exceeding 900 mg/24h on day 9. In two separate experiments involving 52 animals we observed a significant increase in cathepsin D-like activity in cortex (+82.7%), tubules (+109.6%) and glomeruli (+54.7%) isolated from PAN-treated rats killed during marked proteinuria (day 9, mean total urinary protein excretion: 937 +/- 94 mg/24h). This increase was observed whether the activity was expressed per mg of DNA or per mg of protein. Increased cathepsin D-like activity was first observed in cortex and tubules coincident with the onset of proteinurea (day 4, mean total urinary protein excretion: 112 +/- 23 mg/24h). In contrast with the significant elevation of renal cathepsin D-like activity, the activity (nmol/h per mg of protein) of alpha-L-fucosidase (EC 3.2.1.51), a non-proteolytic enzyme, was markedly decreased in the identical samples used for the measurement of cathepsin D-like activity: cortex (-46.4%); tubules (-46.1%); and glomeruli (-38.5%). In addition to changes in renal enzyme activities, PAN-treated rats excreted large amounts of cathepsin D-like activity in their urine (beginning on day 3) compared with nearly undetectable cathepsin D-like activity in the urine from control rats. The significant increases in glomerular and tubular cathepsin D activity may reflect an important role for this enzyme in the pathophysiology associated with PAN-induced nephrotic syndrome.

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Claudin-6 localized in tight junctions of rat podocytes

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.00862.2007 ◽

2008 ◽

Vol 294 (6) ◽

pp. R1856-R1862 ◽

Cited By ~ 21

Author(s):

Linning Zhao ◽

Eishin Yaoita ◽

Masaaki Nameta ◽

Ying Zhang ◽

Lino Munoz Cuellar ◽

...

Keyword(s):

Tight Junctions ◽

Early Stage ◽

Transmembrane Protein ◽

Rat Kidney ◽

Intercellular Junctions ◽

Cell Contact ◽

Puromycin Aminonucleoside ◽

Contact Sites ◽

Stage Of Development ◽

Cell Cell

Tight junctions rarely exist in podocytes of the normal renal glomerulus, whereas they are the main intercellular junctions of podocytes in nephrosis and in the early stage of development. Claudins have been identified as tight junction-specific integral membrane proteins. Those of podocytes, however, remain to be elucidated. In the present study, we investigated the expression and localization of claudin-6 in the rat kidney, especially in podocytes. Western blot analysis and RT-PCR revealed that the neonatal kidney expressed much higher levels of claudin-6 than the adult kidney. Immunofluorescence microscopy showed intense claudin-6 staining in most of the tubules and glomeruli in neonates. The staining in tubules declined distinctly in adults, whereas staining in glomeruli was well preserved during development. Claudin-6 in glomeruli was distributed along the glomerular capillary wall and colocalized with zonula occludens-1. The staining became conspicuous after kidney perfusion with protamine sulfate (PS) to increase tight junctions in podocytes. Immunoelectron microscopy showed that immunogold particles for claudin-6 were accumulated at close cell-cell contact sites of podocytes in PS-perfused kidneys, whereas a very limited number of immunogold particles were detected, mainly on the basal cell membrane and occasionally at the slit diaphragm and close cell-cell contact sites in normal control kidneys. In puromycin aminonucleoside nephrosis, immunogold particles were also found mainly at cell-contact sites of podocytes. These findings indicate that claudin-6 is a transmembrane protein of tight junctions in podocytes during development and under pathological conditions.

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The Morphology of the Rat Kidney Following Folic Acid Administration

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100067996 ◽

1970 ◽

Vol 28 ◽

pp. 200-201

Author(s):

Aline Byrnes ◽

Elsa E. Ramos ◽

Minoru Suzuki ◽

E.D. Mayfield

Keyword(s):

Folic Acid ◽

De Novo ◽

Specific Activity ◽

Rat Kidney ◽

Present Report ◽

Intracellular Localization ◽

Male Rats ◽

Renal Hypertrophy ◽

Electron Microscopic ◽

Biochemical Alterations

Renal hypertrophy was induced in 100 g male rats by the injection of 250 mg folic acid (FA) dissolved in 0.3 M NaHCO3/kg body weight (i.v.). Preliminary studies of the biochemical alterations in ribonucleic acid (RNA) metabolism of the renal tissue have been reported recently (1). They are: RNA content and concentration, orotic acid-c14 incorporation into RNA and acid soluble nucleotide pool, intracellular localization of the newly synthesized RNA, and the specific activity of enzymes of the de novo pyrimidine biosynthesis pathway. The present report describes the light and electron microscopic observations in these animals. For light microscopy, kidney slices were fixed in formalin, embedded, sectioned, and stained with H & E and PAS.

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Biological and Surface Properties of C-Type Virus Particles Produced by Novikoff Ascites Hepatoma Cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100079395 ◽

1977 ◽

Vol 35 ◽

pp. 388-389

Author(s):

D.C. Hixson ◽

J.C. Chan ◽

J.M. Bowen ◽

E.F. Walborg

Keyword(s):

Surface Properties ◽

Ricinus Communis ◽

Rat Kidney ◽

Leukemia Virus ◽

Viral Envelope ◽

Virus Particles ◽

Chemically Induced ◽

Sarcoma Virus ◽

Hepatocarcinoma Cells ◽

Type C

Several years ago Karasaki (1) reported the production of type C virus particles by Novikoff ascites hepatocarcinoma cells. More recently, Weinstein (2) has reported the presence of type C virus particles in cell cultures derived from transplantable and primary hepatocellular carcinomas. To date, the biological function of these virus and their significance in chemically induced hepatocarcinogenesis are unknown. The present studies were initiated to determine a possible role for type C virus particles in chemically induced hepatocarcinogenesis. This communication describes results of studies on the biological and surface properties of type C virus associated with Novikoff hepatocarcinoma cells.Ecotropic and xenotropic murine leukemia virus (MuLV) activity in ascitic fluid of Novikoff tumor-bearing rats was assayed in murine sarcoma virus transformed S+L- mouse cells and S+L- mink cells, respectively. The presence of sarcoma virus activity was assayed in non-virus-producing normal rat kidney (NRK) cells. Ferritin conjugates of concanavalin A (Fer-Con wheat germ agglutinin (Fer-WGA), and Ricinus communis agglutinins I and II (Fer-RCAI and Fer-RCAII) were used to probe the structure and topography of saccharide determinants present on the viral envelope.

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