scholarly journals Postnatal regulation of 15-hydroxyprostaglandin dehydrogenase in the rat kidney

2014 ◽  
Vol 307 (4) ◽  
pp. F388-F395 ◽  
Author(s):  
Ying Liu ◽  
Zhanjun Jia ◽  
Ying Sun ◽  
Li Zhou ◽  
Maicy Downton ◽  
...  
Keyword(s):  
Kidney Development ◽  
Rat Kidney ◽  
Periodic Acid ◽  
Proximal Tubules ◽  
Periodic Acid Schiff ◽  
Qrt Pcr ◽  
Pathological Conditions ◽  
Brush Borders ◽  
Cox 2 ◽  
Established Role

Cyclooxygenase 2 (COX-2) has an established role in postnatal kidney development. 15-Hydroxyprostaglandin dehydrogenase (15-PGDH) is recently identified as an endogenous inhibitor of COX-2, limiting the production of COX-2-derived prostanoids in several pathological conditions. The present study was undertaken to examine the regulation of renal 15-PGDH expression during postnatal kidney development in rats compared with COX-2. qRT-PCR and immunoblotting demonstrated that 15-PGDH mRNA and protein in the kidney were present in neonates, peaked in the second postnatal week, and then declined sharply to very low level in adulthood. Immunostaining demonstrated that at the second postnatal week, renal 15-PGDH protein was predominantly found in the proximal tubules stained positive for Na/H exchanger 3 and brush borders (periodic acid-Schiff), whereas COX-2 protein was restricted to macular densa and adjacent thick ascending limbs. Interestingly, in the fourth postnatal week, 15-PGDH protein was redistributed to thick ascending limbs stained positive for the Na-K-2Cl cotransporter. After 6 wk of age, 15-PGDH protein was found in the granules in subsets of the proximal tubules. Overall, these results support a possibility that 15-PGDH may regulate postnatal kidney development through interaction with COX-2.

scholarly journals Antioxidant Protection against Pathological Mycotoxins Alterations on Proximal Tubules in Rat Kidney

2011 ◽  
Vol 1 (4) ◽  
pp. 118 ◽  
Author(s):  
Awatef Ali ◽  
Susan Abdu
Keyword(s):  
Serum Creatinine ◽  
Ochratoxin A ◽  
Rat Kidney ◽  
Animal Health ◽  
Periodic Acid ◽  
Relative Weight ◽  
Proximal Tubules ◽  
Protective Effects ◽  
Periodic Acid Schiff ◽  
Ultra Structure

Background: Ochratoxin A (OTA) was one of the mycotoxins and received attention worldwide because of the hazard it posed to human and animal health, where the kidney was the primary target organ for OTA toxicity. In the other hand, dates served as a good source of natural antioxidants and could potentially be considered as a functional food.Methods: The study was performed in the department of biology in King Abdulaziz University. Animals were gavage administrated and divided into four groups: first group received (sodium bicarbonate), second group received (289 µg OTA /kg B.W. /day), third group received (1mg Ajwa/kg B.W. / day) and fourth group received (289 µg OTA /kg B.W./day+ 1mg Ajwa /kg B.W. / day). Serum (creatinine - urea) levels were measured in each group at the time of tissue collection, some biopsies were fixed in 10% buffered formalin solution for light microscopy processing stained with Haematoxylin and Eosin (H& E.), Periodic Acid-Schiff (PAS) and Masson´s Trichrome (M.T.).Other biopsies were immediately collected into electron microscopy processing. Results: After 28 days, a significant decrease in body weight, kidney weight and relative weight was detected in OTA treated group. Also, Serum (creatinine-urea) level were elevated. The normal cyto-architecture of proximal tubules were lost exhibiting damaged bruch border, degenerated, binucleated and karyomegalic cells. The most destructed ultra-structure was the mitochondria which severely swollen with disintegrated membranes. In Ajwa Date extract-group the proximal tubules were normal, whereas in Ajwa date extract + OTA -group the severity of the lesions was significantly reduced. Conclusion: The present results indicated that, Ajwa date have protective effects and ameliorated the lesions of Ochratoxin nepherotoxicity which might lead to kidney failure. Key words: Ochratoxin A., Ajwa date, proximal tubules, light –structure, ultra –structure, biochemical analysis, morphometry.

Isolation of a pure suspension of rat proximal tubules

1981 ◽  
Vol 241 (4) ◽  
pp. F403-F411 ◽  
Author(s):  
P. Vinay ◽  
A. Gougoux ◽  
G. Lemieux
Keyword(s):  
Phosphoenolpyruvate Carboxykinase ◽  
Periodic Acid ◽  
Vital Staining ◽  
Staining Technique ◽  
Proximal Tubules ◽  
Periodic Acid Schiff ◽  
Homogeneous Population ◽  
Proximal Tubular ◽  
Percoll Density Gradient ◽  
Enzyme Location

A suspension of cortical tissue fragments prepared by collagenase digestion of renal cortex obtained from fed and chronically acidotic (NH4Cl) rats was separated into four bands on a Percoll density gradient. By microscopic examination, vital staining with trypan blue, and histologic staining technique (periodic acid-Schiff) the F4 band was shown to contain only (greater than 98%) proximal tubules, whereas the F1 band was significantly enriched (70%) with distal tubules contaminated by glomeruli and short segments of proximal tubules. Intra/extracellular ratios for PAH of 15 were measured in the F4 band and of 2 in F1 band. ATP was 1.4 and 2.8 mumol/g in the F4 and F1 bands, respectively, and was stable for at least 60 min. The proximal F4 band was shown to be gluconeogenic (L-glutamine or L-lactate 2.5 mM as substrate) and to adapt to metabolic acidosis. The distal F1 band was shown to be glycolytic (glucose 2.5 mM) with no changes with acid-base status. All fractions were shown to metabolize glutamine, but the metabolic fate of this amino acid was different in proximal and distal structures. A F4/F1 activity ratio for the proximal cytoplasmic phosphoenolpyruvate carboxykinase enzyme of 2.6 and 4.3 was observed in normal and acidotic rats, respectively. In contrast, a F4/F1 ratio of 0.13 and 0.22 was observed for the distal cytoplasmic hexokinase enzyme. This preparation, therefore, allows the metabolism of a homogeneous population of proximal tubular fragments to be studied and can be used to obtain information on enzyme location within the nephron.

The Properties of Soluble Proteins Synthesized by Nephrotic Rat Kidney Microsomes

1972 ◽  
Vol 50 (3) ◽  
pp. 292-298
Author(s):  
V. N. Katiyar ◽  
B. L. Dinh
Keyword(s):  
Gel Electrophoresis ◽  
Rat Kidney ◽  
Periodic Acid ◽  
Rabbit Antiserum ◽  
Periodic Acid Schiff ◽  
Rat Serum ◽  
Tissue Protein ◽  
Schiff Reagent ◽  
Normal Rat ◽  
Kidney Microsomes

The incorporation of 14C-leucine into the microsomal proteins of nephrotic rat kidney was much higher than that in the microsomal proteins of the normal rat kidney. When the newly synthesized microsomal proteins were analyzed by acrylamide gel electrophoresis, it was found that the higher incorporation of 14C-leucine in nephrotic rat kidney was mostly due to an increase in the biosynthesis of a tissue protein which migrated in the electrophoretic zone between serum albumin and transferrin. This protein did not react with rabbit antiserum to normal rat serum and was stained with periodic acid – Schiff reagent. It was believed to be excreted in the urine of nephrotic rats in great quantity and was easily identified as a distinct band in electrophoregrams of urine from these rats.

Effect of ischemic acute renal damage on the expression of COX-2 and oxidative stress-related elements in rat kidney

2007 ◽  
Vol 292 (5) ◽  
pp. F1364-F1371 ◽  
Author(s):  
Sandra Villanueva ◽  
Carlos Céspedes ◽  
Alexis A. González ◽  
Carlos P. Vio ◽  
Victoria Velarde
Keyword(s):  
Oxidative Stress ◽  
Inflammatory Responses ◽  
Rat Kidney ◽  
Smooth Muscle Actin ◽  
Periodic Acid ◽  
Heme Oxygenase 1 ◽  
Transcriptional Responses ◽  
Bradykinin B2 Receptor ◽  
Cox 2 ◽  
And Oxidative Stress

Acute renal failure (ARF) is a clinical syndrome characterized by deterioration of renal function over a period of hours or days. The principal causes of ARF are ischemic and toxic insults that can induce tissue hypoxia. Transcriptional responses to hypoxia can be inflammatory or adaptive with the participation of the hypoxia-inducible factor 1α and the expression of specific genes related to oxidative stress. The production of peroxynitrites and protein nitrotyrosylation are sequelae of oxidative stress. In several clinical and experimental conditions, inflammatory responses have been related to cyclooxygenase (COX)-2, suggesting that its activation might play an important role in the pathogenesis and progression of nephropathies such as ARF. In the kidney, renin and bradykinin participate on the regulation of COX-2 synthesis. With the hypothesis that in ARF there is an increase in the expression of agents involved in adaptive and inflammatory responses, the distribution pattern and abundance of COX-2, its regulators renin, kallikrein, bradykinin B2 receptor, and oxidative stress elements, heme oxygenase-1 (HO-1), erythropoietin (EPO), inducible nitric oxide synthase (iNOS), and nitrotyrosylated residues were studied by immunohistochemistry and immunoblot analysis in rat kidneys after bilateral ischemia. In kidneys with ARF, important initial damage was demonstrated by periodic acid-Schiff staining and by the induction of the damage markers α-smooth muscle actin and ED-1. Coincident with the major damage, an increase in the abundance of EPO, HO-1, and iNOS and an increase in renin and bradykinin B2 receptor were observed. Despite the B2 receptor induction, we observed an important decrease in COX-2 in the ischemic-reperfused kidney. These results suggest that COX-2 does not participate in inflammatory responses induced by hypoxia.

scholarly journals Localization of d-myoinositol 1:2-cyclic phosphate 2-phosphohydrolase in rat kidney

1972 ◽  
Vol 130 (1) ◽  
pp. 229-238 ◽  
Author(s):  
N. Clarke ◽  
R. M. C. Dawson
Keyword(s):  
Alkaline Phosphatase ◽  
Rat Kidney ◽  
Gradient Centrifugation ◽  
Subcellular Fractionation ◽  
Proximal Tubules ◽  
Outer Cortex ◽  
Inositol 1 ◽  
Brush Borders ◽  
Cyclic Phosphate ◽  
Pars Convoluta

1. On subcellular fractionation of rat kidney homogenates by differential and density-gradient centrifugation, the bulk of the inositol 1:2-cyclic phosphate 2-phosphohydrolase activity remains with the alkaline phosphatase activity, suggesting localization in the brush borders of the proximal tubules. 2. Histochemical studies with a medium containing inositol 1:2-cyclic phosphate and Escherichia coli phosphomonoesterase show Gomori staining around the brush borders of the proximal tubules in the outer cortex only. 3. Serial sections across the kidney from cortex perimeter to papilla suggest that the inositol 1:2-cyclic phosphate 2-phosphohydrolase has a limited distribution along the proximal tubule of the nephron, probably being limited to the pars convoluta, whereas the alkaline phosphatase extends along the pars recta.

Membrane repolarization is delayed in proximal tubules after ischemia-reperfusion: possible role of microtubule-organizing centers

2003 ◽  
Vol 285 (2) ◽  
pp. F230-F240 ◽  
Author(s):  
Flavia A. Wald ◽  
Yolanda Figueroa ◽  
Andrea S. Oriolo ◽  
Pedro J. I. Salas
Keyword(s):  
Apical Membrane ◽  
Rat Kidney ◽  
Ischemia Reperfusion ◽  
Proximal Tubules ◽  
Microtubule Organizing Centers ◽  
Apical Domain ◽  
Brush Borders ◽  
Chemical Anoxia

We have previously shown that microtubule-organizing centers (MTOCs) attach to the apical network of intermediate filaments (IFs) in epithelial cells in culture and in epithelia in vivo. Because that attachment is important for the architecture of microtubules (MTs) in epithelia, we analyzed whether chemical anoxia in LLC-PK1 and CACO-2 cells or unilateral ischemia-reperfusion in rat kidney (performed under fluorane anesthesia) had an effect on the binding and distribution of MTOCs. In culture, we found that chemical anoxia induces MTOC detachment from IFs by morphological and biochemical criteria. In reperfused rat proximal tubules, noncentrosomal MTOCs were fully detached from the cytoskeleton and scattered throughout the cytoplasm at 3 days after reperfusion, when brush borders were mostly reassembled. At that time, MTs were also fully reassembled but, as expected, lacked their normal apicobasal orientation. Two apical membrane markers expressed in S2 and S3 segments were depolarized at the same stage. At 8 days after reperfusion, membrane polarity, MTOCs, and MTs were back to normal. Na+-K+-ATPase was also found redistributed, not to the apical domain but rather to an intracellular compartment, as described by others (Alejandro VS, Nelson W, Huie P, Sibley RK, Dafoe D, Kuo P, Scandling JD Jr., and Myers BD. Kidney Int 48: 1308–1315, 1995). The prolonged depolarization of the apical membrane may have implications in the pathophysiology of acute renal failure.

Low endogenous glucocorticoid allows induction of kidney cortical cyclooxygenase-2 during postnatal rat development

2004 ◽  
Vol 286 (1) ◽  
pp. F26-F37 ◽  
Author(s):  
Kirsten Madsen ◽  
Jane Stubbe ◽  
Tianxin Yang ◽  
Ole Skøtt ◽  
Sebastian Bachmann ◽  
...  
Keyword(s):  
Kidney Development ◽  
Collecting Duct ◽  
Rat Kidney ◽  
Plasma Concentrations ◽  
Cyclooxygenase 2 ◽  
Kidney Cortex ◽  
Thick Ascending Limb ◽  
Corticosterone Concentration ◽  
Rat Kidney Cortex ◽  
Cox 2

In postnatal weeks 2–4, cyclooxygenase-2 (COX-2) is induced in the rat kidney cortex where it is critically involved in final stages of kidney development. We examined whether changes in circulating gluco- or mineralocorticosteroids or in their renal receptors regulate postnatal COX-2 induction. Plasma corticosterone concentration peaked at birth, decreased to low levels at days 3- 13, and increased to adult levels from day 22. Aldosterone peaked at birth and then stabilized at adult levels. Gluco- and mineralocorticoid receptor (GR and MR) mRNAs were expressed stably in kidney before, during, and after COX-2 induction. 11β-Hydroxysteroid dehydrogenase 2 was induced shortly after birth and was widely distributed in the whole collecting duct system in the suckling period and then returned to an adult pattern. Supplementation with corticosterone (20 mg·kg-1·day-1) or GR-specific dexamethasone (1 mg·kg-1·day-1) during low endogenous corticosterone suppressed renal COX-2 mRNA and protein and led to a restricted distribution of COX-2 immunolabeling. The ability of glucocorticoids to affect COX-2 was reflected in colocalization of GR-α and COX-2 immunoreactivity and mRNAs in thick ascending limb of Henle's loop. The MR antagonist potassium canrenoate (20 mg·kg-1·day-1) enhanced COX-2 expression from days 5 to 10, but low MR-specific concentrations of DOCA (1 mg·kg-1·day-1) had no effect on COX-2. Renomedullary interstitial cells expressed GR-α and COX-2. Dexamethasone suppressed COX-2 in these cells. Thus low plasma concentrations of corticosterone allowed for cortical and medullary COX-2 induction during postnatal kidney development. Increased circulating glucocorticoid in the postnatal period may damage late renal development through inhibition of COX-2.

scholarly journals Differential expression of gap junction mRNAs and proteins in the developing murine kidney and in experimentally induced nephric mesenchymes

Development ◽  
1992 ◽  
Vol 115 (3) ◽  
pp. 827-837
Author(s):  
K. Sainio ◽  
S.F. Gilbert ◽  
E. Lehtonen ◽  
M. Nishi ◽  
N.M. Kumar ◽  
...  
Keyword(s):  
Gap Junction ◽  
Kidney Development ◽  
Rat Kidney ◽  
Physiological Activity ◽  
Proximal Tubules ◽  
Small Subset ◽  
Gene Products ◽  
Intrauterine Development ◽  
Beta 2 ◽  
Experimentally Induced

The expression of three gap junction (GJ) proteins, alpha 1 (Cx43), beta 1 (Cx32), and beta 2 (Cx26), and their transcripts were examined during the ontogeny of the mouse and rat kidney. These proteins were expressed in two non-overlapping patterns. The alpha 1 GJ protein was first observed in mesenchymal cells in the 12-day mouse kidney. By day 14 and thereafter, the alpha 1 protein was detected in the transient S-shaped bodies, but not in the podocytes of the maturing glomeruli. After birth the antigen was retained in a small subset of secretory tubules. The beta 1 and beta 2 GJ proteins were similar in their developmental patterns. They were first detected in a small subset of secretory tubules in the subcortical zone of day 17 embryos. These tubules were identified by immunohistochemical markers to be proximal. At birth, practically all proximal tubules expressed the two antigens. This analysis of GJ proteins was consistent with the results of S1 nuclease protection assays showing that, while the alpha 1 mRNA appeared early during kidney development and declined around birth, the two beta mRNAs appeared later and became intensified during the last days of intrauterine development. In experimentally induced metanephric mesenchymes, a transient expression of the alpha 1 GJ protein was seen during the segregation of the tubular anlagen. beta 1 and beta 2 GJ proteins were not detected in such induced mesenchymes cultivated up to 7 days. These observations provide evidence for the cell-specific utilization of different GJ genes during different stages of kidney organogenesis. The alpha 1 gene is activated during the early segregation of the secretory tubule and might contribute to its compartmentalization, while the beta 1 and beta 2 gene products are not detected until advanced stages of development. The latter gene products might be correlated with the physiological activity of the proximal tubules in vivo, as they are not expressed in experimentally induced tubules detectable with markers for proximal tubules.

scholarly journals Release of intestinal surface-membrane glycoproteins associated with enzyme activity by brief digestion with papain

1971 ◽  
Vol 121 (5) ◽  
pp. 781-789 ◽  
Author(s):  
Gordon G. Forstner
Keyword(s):  
Brush Border ◽  
Gradient Centrifugation ◽  
Periodic Acid ◽  
Polyacrylamide Gel Electrophoresis ◽  
Surface Membrane ◽  
Membrane Glycoproteins ◽  
Periodic Acid Schiff ◽  
Brush Borders ◽  
And Function ◽  
Sepharose 4B

Rat intestinal surface-membrane glycoproteins were labelled by intraperitoneal injection of [1-14C]glucosamine 4h before the animals were killed. At this time, density-gradient centrifugation of disrupted brush borders indicated that glycoprotein radioactivity was distributed identically with sucrase, a plasma-membrane marker. Labelled brush borders were digested by papain for brief time-intervals known to release surface-enzyme particles without disruption of the unit membrane. Digestion for 5min released 90% of the surface sucrase, and almost one-half of the brush-border glycoprotein and label. On Sepharose 4B column chromatography most of the glycoprotein and label emerged as a single peak. This peak contained the most actively labelled glycoprotein in the brush border and was closely associated with maltase, sucrase, β-naphthylamidase and alkaline phosphatase. The peak was partially resolved on polyacrylamide-gel electrophoresis into three bands. Each band contained a distinctive enzyme or enzyme pair, and was labelled by [1-14C]glucosamine. No periodic acid–Schiff-negative protein was observed in the peak material. Glycoproteins susceptible to brief digestion with papain are therefore closely linked to released surface-enzyme particles. Intestinal surface glycoproteins are heterogeneous with respect to molecular weight, electrophoretic mobility and function.

scholarly journals Immunohistochemical Localization of Peroxisomal Enzymes in Developing Rat Kidney Tissues

1998 ◽  
Vol 46 (10) ◽  
pp. 1161-1173 ◽  
Author(s):  
Kohei Johkura ◽  
Nobuteru Usuda ◽  
Yan Liang ◽  
Ayami Nakazawa
Keyword(s):  
Kidney Development ◽  
Rat Kidney ◽  
Immunoblot Analysis ◽  
Immunohistochemical Localization ◽  
Proximal Tubules ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
Hydroxyacyl Coa Dehydrogenase ◽  
Developing Rat ◽  
Neonatal Kidney

We studied the developmental changes in the localization of peroxisome-specific enzymes in rat kidney tissues from embryonic Day 16 to postnatal Week 10 by immunoblot analysis and immunohistochemistry, using antibodies for the peroxisomal enzymes catalase, d-amino acid oxidase, L-α-hydroxyacid oxidase (isozyme B), and enoyl-CoA hydratase/3-hydroxyacyl-CoA dehydrogenase bifunctional protein. Peroxisomal enzymes were detected in the neonatal kidney by immunoblot analysis and their amount increased with kidney development. By light microscopic immunohistochemistry, they were first localized in a few proximal tubules in the juxtamedullary cortex of 18-day embryos. The distribution of proximal tubules positive for them expanded towards the superficial cortex with development. The full thickness of the cortex became positive for the staining by 14 days after birth. Peroxisomes could be detected by electron microscopy in structurally immature proximal tubules in 18-day embryos. Their size increased and the ultrastructure of subcompartments became clear with continuing development of proximal tubules. These results show that peroxisomal enzymes appear in the immature proximal tubules in the kidney of embryos and that the ultrastructure of the peroxisomes and localization of the peroxisomal enzymes develop along with the maturation of proximal tubules and kidney tissues.

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