Normal Dog Kidney Research Articles

Expression of Phosphatonin-Related Genes in Sheep, Dog and Horse Kidneys Using Quantitative Reverse Transcriptase PCR.

Simple SummaryTraditionally, it has been thought that control of body phosphorus was secondary to the tighter control of calcium. However, over the last 20 years, an extensive system for control of body phosphorus by proteins called phosphatonins has been shown to exist. Most research on phosphatonins has been done in rat or mouse models. This paper looks at whether important proteins and phosphorus channels in the phosphatonin pathways are present in the kidneys of dogs, horses and sheep. The results showed that all of the components of the phosphatonin system are present in these species, but that there are species differences in which protein or channel is most common, and in the relationships between the proteins and channels. This research is important because the phosphatonin system is involved in the progression of chronic kidney disease in humans and animals, and differences in the systems between animal species may affect treatment of chronic kidney disease.The aim of this preliminary study was to determine the relative expression of phosphatonin pathway-related genes in normal dog, sheep and horse kidneys and to explore the relationships between the different genes. Kidneys were collected post-mortem from 10 sheep, 10 horses and 8 dogs. RNA was extracted, followed by reverse transcriptase quantitative polymerase chain reaction for fibroblast growth factor receptor 1 IIIc (FGFR1IIIC), sodium-phosphate co-transporter (NPT) 1 (SLC17A1), NPT2a (SLC34A1), NPT2c (SLC34A3), parathyroid hormone 1 receptor (PTH1R), klotho (KL), vitamin D receptor (VDR), 1a-hydroxylase (CYP27B1) and 24-hydroxylase (CYP24A1). NPT2a was highly expressed in the dog kidneys, compared with those of the horses and sheep. NPT1 had greatest expression in horses and sheep, although the three different NPTs all had relatively similar expression in sheep. There was little variability in FGFR1IIIc expression, particularly in the dogs and horses. FGFR1IIIc expression was negatively correlated with NPT genes (except NPT2a in sheep), while NPT genes were all positively correlated with each other. Unexpectedly, klotho was positively correlated with NPT genes in all three species. These results provide the basis for further research into this important regulatory system. In particular, species differences in phosphatonin gene expression should be considered when considering the pathogenesis of chronic kidney disease.

Displasia renal em cães: estudo retrospectivo (2008-2013)

Background: Renal dysplasia, which has been reported in some dogs and human patients, refers to a developmental disorder of renal parenchyma due to imperfect inductive interaction between the mesonephric duct and the metanephric blastemal. In dogs, the characteristic histological findings on which diagnosis is based include (1) persistent metanephric ducts surrounded by primitive mesenchyme, (2) fetal or immature glomeruli, (3) fetal or immature tubules, and (4) anomalous presence of interstitial fibrous tissue. The aim of this study was to report the major pathological and immunohistochemical features of nine young dogs necropsied with renal dysplasia.Cases: The necropsy files from the Laboratório de Patologia Veterinária (LPV) of the Universidade Federal do Mato Grosso (UFMT) were reviewed between the years 2008 and 2013. Dogs diagnosed with chronic kidney failure and macroscopic and histopathological renal lesions consistent with renal dysplasia were selected. Kidney fragments in paraffin blocks were cut and stained with hematoxylin and eosin and by immunohistochemistry (IHC) using anti-vimentin and anti-cytokeratin monoclonal antibodies. The staining was considered positive for the presence of at least one renal cell marked with brown cytoplasmic staining clear and unambiguous. A total of 787 necropsies of dogs were performed. Of these, 64 had a clinical diagnosis of chronic renal failure of which 9 were classified as renal dysplasia. The age of the dogs ranged from 3 months to 2 years. Clinical signs were characterized by anorexia and non-regenerative anemia in 88.9%, vomiting 66.7%, dehydration 55.6%, uremia 55.6%, convulsion 33.4%, abdominal pain 22.3% and diarrhea in 11.2% of cases. At necropsy the main macroscopic changes in the kidneys were external surface with pale staining, in 6 of 9 dogs necropsied. Additionally, 5 dogs, cystic cavities of various sizes from 0.1 to 5 cm in diameter, diffusely distributed in the renal subcapsular surface and cutting were observed. Of the 64 dogs with diagnosis of chronic renal failure, 14.06% had dysplastic kidney changes, characterized by dilatation of Bowman’s space, glomerular and tubular atrophy, immature glomeruli and tubules, lymphocytic interstitial inflammation and fibrosis. In the dogs with renal dysplasia, it was observed that the tubular structures showed marked glomerular labeling for vimentin. Moreover, the kidneys of normal dogs showed weak or absent for marking tubes and glomerular structures for vimentin, and strong staining for cytokeratin in tubular cells, glomerular cells and collecting ducts.Discussion: Renal morphological damage observed in these nine dogs less than two years old, that contained degenerative and inflammatory changes, fibrosis and glomerular atrophy, but mostly immature glomeruli and tubules were associated with atypical renal dysplasia. This condition develops when the urethral diverticulum and metanephric blastoma not properly form in the embryonic stage, resulting in abnormal metanephric differentiation and formation of structures that do not recapitulate the normal nephrogenesis. Abnormal kidney function in young animals caused chronic renal failure resulting in death. The results of immunohistochemical observed in this study can complement the diagnosis of renal dysplasia. The morphological normality can be observed in the proportion of mesenchymal and epithelial tissue in the different structures of the kidney by staining with anti-cytokeratin and anti-vimentin. In the present study, there was moderate to strong labeling of vimentin in glomerular and tubular structures being dysplastic kidney would be expected to occur in normal dogs predominant staining with cytokeratin. Based on the clinic, pathological and immunohistochemical findings it is concluded that the animals developed renal dysplasia.

Participação da excreção renal de cálcio, fósforo, sódio e potássio na homeostase em cães sadios e cães com doença renal crônica

Na doença renal crônica (DRC) a manutenção da homeostase de água e sódio é o primeiro problema a ser contornado pelo organismo e com o agravamento das lesões renais surgem outros problemas graves relacionados à homeostase de cálcio e fósforo. O presente estudo tem por escopo avaliar a excreção renal de cálcio, fósforo, sódio e potássio, e o perfil sérico destes eletrólitos em cães normais e em cães com DRC naturalmente adquirida. Foram avaliados três grupos de cães adultos, machos ou fêmeas, de raças variadas. Animais normais compuseram o grupo controle (G1) e os cães com DRC foram distribuídos em dois grupos de acordo com os estágios de comprometimento da função renal (G2 e G3, respectivamente, estágios 1-2 e estágios 3-4, descritos pela IRIS 2006 staging CKD). Os cães do G3 apresentaram aumento das concentrações séricas de cálcio ionizado e fósforo, além de diminuição da concentração sérica de sódio. Quanto à excreção renal dos eletrólitos analisados, os animais dos grupos G1 e G2 apresentaram diminuição de carga filtrada e aumento de excreção fracionada, mas as excreções urinárias não variaram significativamente. Os resultados são indicativos de que os rins de cães com DRC podem manter a excreção urinária dos eletrólitos em valores se melhantes aos dos normais. O mecanismo envolve aumento da excreção fracionada na medida em que haja diminuição da filtração glomerular. Esse processo de compensação, entretanto, pode perder a eficiência nos estágios mais avançados da enfermidade no que se refere à manutenção das concentrações séricas de fósforo e sódio.

Canine prostate stimulates osteoblast function using the endothelin receptors.

Bone metastases are common in humans and dogs with late-stage prostate cancer. A unique feature of prostate cancer metastases is new bone formation at metastatic sites ("osteoblastic metastases"). Many carcinomas that metastasize to bone cause bone destruction, not new bone formation. The mechanisms by which prostate cancer induces bone formation at sites of bone metastasis are not well understood. We hypothesized that stimulation of osteoblasts by prostate tissue at metastatic sites was due to the paracrine actions of growth factors produced by prostate epithelial cells. We have previously shown that normal canine prostate tissue induced new bone formation when implanted adjacent to the calvarium of nude mice. To complement this in vivo model, we developed an in vitro system of prostate-stimulated osteoblast function to investigate mechanisms of prostate-induced new bone formation. We found that treatment of cultured rat calvaria for 24 hr with proteins from normal dog prostate stimulated alkaline phosphatase activity in a dose-dependent manner 4-6 fold compared to controls. Stimulation began approximately 8 hr after treatment, and was diminished after 72 hr. Calvaria treated with homogenates of normal dog salivary gland, kidney, bladder, and muscle did not increase ALP activity. Pretreatment of the calvaria for 1 hr with endothelin antagonists, but not anti-parathyroid hormone-related protein (PTHrP) antibody or indomethacin, abrogated the stimulation of ALP. Our results indicated that osteoblast activation by canine prostate occurs via an endothelin-dependent mechanism, and that PTHrP or prostaglandin synthase-mediated pathways are likely not involved. This is a reliable, reproducible assay for determining the roles of molecules important in the activation of osteoblasts by the prostate.

Hepatocyte growth factor ameliorates renal hemodynamic disorder after ischemia/reperfusion

Ischemic injury of the transplanted kidney is one of the causes of reduced graft survival. The purpose of the present experiment was to examine whether hepatocyte growth factor (HGF) would improve acute renal hemodynamic recovery immediately after cold ischemia. Addition of HGF to the preservation solution during 3 h cold ischemia of dog kidney accelerated both recovery of renal blood flow and glomerular filtration rate (GFR). It is suggested that HGF may be useful for preservation of excised kidney for transplantation. As intrarenal arterial infusion of HGF in normal dog kidney had no effects on renal hemodynamics, mechanisms other than direct vasodilator action of HGF appear to be operating in the protection.

Ultra-sonografia de características lineares e estimativas do volume de rins de cães

Obtiveram-se as medidas lineares de rins de cães adultos, clinicamente normais, por meio da ultra-sonografia, e estimou-se o volume dos rins de 35 cães (18 fêmeas e 17 machos) sem raça definida e com idade entre dois e seis anos. Os animais foram divididos em três grupos de acordo com o peso corporal (3,1 a 10,0 kg; 10,1 a 20,0 kg e 20,1 a 45,0 kg). Os planos de secção sagital, dorsal e transversal foram utilizados para obtenção das medidas lineares de comprimento (C), largura (L) e espessura (E). O volume renal foi calculado a partir das medidas lineares utilizando-se a fórmula para o volume de um elipsóide. Os valores médios encontrados para os três grupos foram, respectivamente: comprimento (4,73; 6,09 e 7,13cm), largura (2,81; 3,44 e 3,95cm), espessura (2,43; 3,00 e 3,58cm) e volume (17,98; 33,36 e 53,81cm³). As dimensões dos rins esquerdo e direito e de machos e fêmeas foram equivalentes. Observaram-se correlações positivas entre todas as medidas renais e peso corporal.

Molecular cloning, expression and tissue distribution of canine lipopolysaccharide (LPS)-binding protein

The plasma lipopolysaccharide (LPS)-binding protein (LBP) plays an important role in infection caused by Gram-negative bacteria. It is markedly increased during acute-phase responses. In this study, we cloned the full length of canine LBP cDNA and determined its amino-acid sequence and its expression in several canine tissues. The isolated LBP cDNA contained a 1443-bp coding region, which encodes a 25-amino-acid signal peptide, a 456-amino-acid mature LBP and a stop codon, and a 3′-noncoding region containing a TATTTAT motif, which is probably involved in the degradation and/or suppression of mRNA translation. The amino-acid sequence of the mature canine LBP showed 78%, 66% and 67% identity with that of human, rat and rabbit LBPs, respectively. In transient expression assays, canine and human LBPs accelerated the production of tumor necrosis factor- α induced by LPS in human monocytes. Northern blot analysis showed that LBP mRNA is mainly expressed in the liver and kidneys of normal dogs. In situ hybridization analysis revealed that the canine LBP mRNA is mainly located in hepatocytes and in epithelial cells of the proximal urinary tubules of the kidneys. These findings suggest that LBP is produced in organs readily exposed to LPS, where it probably plays an important role in bacterial infections, particularly in those occurring after major surgery.

Non-steroidal anti-inflammatory drug-induced renal failure: a brief review of the role of cyclo-oxygenase isoforms.

Non-steroidal anti-inflammatory drugs are efficacious treatments for rheumatoid arthritis and osteoarthritis. However, an adverse effect of treatment with non-steroidal anti-inflammatory drugs is acute renal failure, particularly in a subset of patients that are in a state of effective volume depletion. The frequency of this side-effect in the general treated population is not known, but is probably less than 1% per year. Non-steroidal anti-inflammatory drugs act by inhibiting the synthesis of prostaglandins, which are important mediators of renal function. In the volume-depleted state prostaglandins may counter the vasoconstriction associated with the activation of the renin-angiotensin system. Cyclooxygenase is the rate-limiting enzyme involved in the synthesis of prostaglandins. Cyclooxygenase exists in two forms: a constitutive form (cyclooxygenase-1) and an inducible form (cyclooxygenase-2), which is associated with inflammation. Non-steroidal anti-inflammatory drugs are non-specific inhibitors of both forms of cyclooxygenase. New data are emerging regarding the role of cyclooxygenase-2 in the control of renal function. In normal rat and dog kidney, cyclooxygenase-2 is sparsely expressed in the macula densa, but expression is upregulated when animals are volume depleted. This review explores the possible role of cyclooxygenase-2 in the maintenance of normal renal function in volume depleted states.

Alterations in F-actin distribution in cells treated with melatonin.

One of the possible pathways of action of melatonin is its effect on the cytoskeleton. In this work we looked for alterations in the cytoskeleton of cells treated with melatonin at physiological concentrations. T-47D, Hs-578T (human breast carcinoma cell lines), and MDCK (normal dog kidney) cells were maintained in MCDB 153 supplemented with 1% fetal bovine serum (FBS), or in Dulbecco's modified Eagle's medium (DMEM) supplemented with 5% FBS and treated with melatonin (10(-9) M or 10(-10) M) for 2 and 5 days, with or without 10(-8) M estradiol. F-actin was stained with phalloidin-fluorescein isothiocyanate (FITC). Cytokeratin 19 and beta-tubulin filaments were detected with specific monoclonal antibodies and secondary antibodies bound to FITC. Melatonin-treated T-47D cells observed in a transmission electronic microscope (TEM) showed an irregular nuclear shape and intermediate filaments disposed around the nucleus, which was not observed in control cells. Immunofluorescence analysis of cytokeratin filaments did not show significant differences between their distribution in control and treated cells. Melatonin did not induce significant alterations in cytokeratin filaments of T-47D, Hs578T or MDCK cells in DMEM and MCDB 153, or T-47D cells in DMEM. Melatonin induced the derangement of F-actin both in T-47D and MDCK cells kept in MCDB 153. The same was not observed when estradiol was also present. We did not observe significant alterations in the distribution of F-actin in T-47D or Hs-578T cells grown in DMEM. In DMEM, melatonin-treated MDCK cells were more elongated, with a slight concentration of F-actin on the cell boundary. Melatonin induced very slight alterations in microtubule organization of all cell lines studied.

Coordinate Gene Expression of the α3, α4, and α5 Chains of Collagen Type IV: EVIDENCE FROM A CANINE MODEL OF X-LINKED NEPHRITIS WITH A COL4A5 GENE MUTATION

Canine X-linked hereditary nephritis is an animal model for human X-linked hereditary nephritis with a premature stop codon in the alpha5(IV) gene of collagen type IV. We used this model to examine the other alpha(IV) chains at the mRNA and protein level in the kidney, since in human X-linked hereditary nephritis, the alpha3(IV) and alpha4(IV) chains are often absent from the glomerular basement membrane, although both are encoded by autosomal genes. cDNA probes for the alpha1(IV)-alpha6(IV) chains were generated from normal dog kidney using the polymerase chain reaction. Sequences were >/=88% identical at the DNA level and >/=92% identical at the protein level to the respective human alpha(IV) chains. By Northern analysis, transcripts for the alpha1(IV), alpha2(IV), and alpha6(IV) chains were detected at comparable levels in both normal and affected male dog kidney RNA. As previously shown, the transcript for the alpha5(IV) chain was reduced to approximately 10% of normal. Unexpectedly, the alpha3(IV) and alpha4(IV) transcripts were both decreased >/=77% in affected male dog kidney, suggesting a mechanism coordinating the expression of these three basement membrane components. The NC1 domain of collagen type IV isolated from normal dog glomeruli was positive for the alpha3(IV), alpha4(IV), and alpha5(IV) chains by Western blotting. In contrast, in the NC1 domain isolated from affected dog glomeruli, these three chains were not detectable, except for a trace of alpha3(IV) dimer. In X-linked hereditary nephritis, the absence of the alpha3(IV) and alpha4(IV) chains from glomerular basement membrane may reflect factors acting at the transcriptional and/or translational level in addition to the protein assembly level.

Renal brush border membrane lipid composition in Basenji dogs with spontaneous idiopathic Fanconi syndrome

To comprehend the renal defect underlying idiopathic Fanconi syndrome in the Basenji dog, we have focused on delineating the lipid profiles of renal brush border membranes isolated from affected and normal Basenji dogs to establish any physical or compositional changes underlying previously observed transport and membrane-fluidity changes. The lipid composition was studied with respect to total lipid, cholesterol, and phospholipid content, cholesterol to phospholipid ratio, distribution of the major phospholipid classes, and fatty acid composition. Total phospholipid of the isolated renal brush border membranes from Fanconi syndrome dogs analyzed by 31P nuclear magnetic resonance showed no difference compared with that of normal dogs. Examination of total fatty acids in both membranes using gas-liquid chromatography analysis of fatty acid methyl esters showed no difference in the mole percents of the major fatty acids. Our data suggest that changes in bulk membrane fluidity of the Fanconi syndrome dog renal brush border as measured by 1,6-diphenyl-1,3,5-hexatriene cannot be attributed to phospholipid and fatty acid compositional change. In the membranes isolated from affected dog kidney, the cholesterol content determined by gas-liquid chromatography analysis was 66 mol% higher than in membranes isolated from normal dog kidney. This correlates with the higher cholesterol to phospholipid molar ratio of 0.82 ± 0.08 in the affected animal as compared with 0.58 ± 0.04 in the normal. Cholesterol content and its microdomain in the membrane bilayer may be important in modulating transport functions. Increased membrane cholesterol content may affect the conformational motility of membrane transport proteins and thus affect their function.

Identifying acoustic scattering sources in normal renal parenchyma in vitro by varying arterial and ureteral pressures

Ultrasonic backscatter properties of normal dog kidney parenchyma are examined in vivo to determine sources of acoustic scattering. We systematically varied the renal perfusion and ureteral pressures to obtain detailed information about scattering sources that could not be seen under in vitro conditions. These data suggest that in normal parenchyma the principal sources of backscatter are Bowman's capsule at low frequencies (2.5–5.0 MHz) and glomerular arterioles at high frequencies (5.0–15.0 MHz). We found that the integrated backscatter coefficient (IBC) in normally perfused kidney cortex is approximately half that measured in the ischemic organ at all frequencies. Ischemia was found to reduce scatterer size estimates ( D) by 10% at low frequencies and increase D 54% at high frequencies. Acute obstruction of the kidney, under diuresis, produced an 11% increase in D at low frequencies, and no significant change in D at high frequencies. These variations in backscatter measurements are explained in terms of changes in the microscopic anatomy of the kidney.

Plasma glutamine and renal ammoniagenesis in dogs with chronic metabolic acidosis.

The purpose of this investigation was to determine whether the rate of glutamine metabolism in the kidneys of normal dogs and dogs with chronic metabolic acidosis was influenced by the plasma glutamine concentration. Because glutamine is a major renal energy fuel, results were examined at a constant rate of energy or ATP turnover [i.e., per 100 ml glomerular filtration rate (GFR)]. Glutamine extraction per 100 ml GFR was directly proportional to the filtered load of glutamine in normal and acidotic dogs. The slope depicting this relationship was parallel to the filtered load of glutamine; however, in normal dogs it was lower and in acidotic dogs it exceeded the filtered load by approximately 22 mumol/100 ml GFR. With respect to the fate of the nitrogens of the glutamine extracted, alanine and ammonium were produced in normal dogs at a rate nearly equivalent to that of glutamine extracted, whereas ammonium production was almost twofold greater than the rate of glutamine extraction during acidosis. There was a relatively small but constant alanine release over the entire range of plasma glutamine concentrations in these dogs. Furthermore, infusion of glutamine to raise the plasma glutamine concentration twofold during acidosis resulted in an increased rate of glutamine extraction and ammonium production equal to that predicted from the increase in filtered load of glutamine. Therefore, variations of circulating glutamine concentration within the physiological range seem to have an important influence on the steady-state rate of renal glutamine metabolism in normal dogs and in dogs with chronic metabolic acidosis.

Atriopeptin III increases cGMP in glomeruli but not in proximal tubules of dog kidney.

Atriopeptin III is natriuretic, diuretic, and phosphaturic in the intact dog and relaxes vascular and nonvascular smooth muscle in vitro. The mechanism of the renal effects of atriopeptin remains ill defined but may include hemodynamic alterations and/or direct tubular effects. Since many peptide hormones act through changes in cyclic nucleotide formation, we examined the effect of synthetic atriopeptins on cAMP and cGMP accumulation in isolated glomeruli and proximal tubules from normal dog kidney. Synthetic atriopeptin produced a dose-related increase in cGMP in glomeruli with a Kact of 200 +/- 19 nM and a maximal activity of 64.1 +/- 6.3 pmol cGMP X mg prot-1 X 5 min-1. In the presence of 4 mM Mn2+ Kact was reduced to 19 +/- 2 nM with no change in maximal cGMP production (66.2 +/- 11.8 pmol cGMP X mg prot-1 X 5 min-1). Atriopeptin (1 microM) did not increase cGMP accumulation above basal levels in proximal tubule segments. The cAMP production of neither glomeruli nor proximal tubules was stimulated by atriopeptin. These studies demonstrate a dose-dependent stimulation of guanylate cyclase by atriopeptin in glomeruli but not in proximal tubule segments. In addition, atriopeptin neither stimulates nor inhibits cAMP accumulation in glomeruli or proximal tubules. These data suggest a major role for the renal effects of atriopeptin at the glomerular level, possibly through hemodynamic effects or changes in the glomerular ultrafiltration coefficient.(ABSTRACT TRUNCATED AT 250 WORDS)

Uptake of Pi in basolateral vesicles after release of unilateral ureteral obstruction.

We previously demonstrated Na+-dependent 32Pi uptake in basolateral membrane vesicles from kidneys of normal dogs. We postulated that this reflects Na+-dependent Pi transport across the basolateral membrane into the proximal tubular cell in vivo that subserves cellular needs for Pi. To ascertain whether Pi transport across the basolateral membrane was enhanced in a condition in which delivery of Pi across the luminal membrane would be markedly decreased, we measured Na+-dependent 32Pi uptake in basolateral vesicles isolated from postobstructed and control kidneys of dogs following release of unilateral ureteral obstruction. Clearance of creatinine and filtered load of Pi were significantly decreased in postobstructed compared with control kidneys. Na+-dependent 32Pi uptake was increased in basolateral vesicles from postobstructed compared with control kidneys, as reflected by increased initial rate of uptake and an increased overshoot of 32Pi transport. Our findings are consistent with an adaptation resulting in increased transport of Pi across the basolateral membrane of the proximal tubular cell in unilateral ureteral obstruction. This process may provide required Pi to the cell under conditions in which decreased Pi is available for transport across the brush border membrane.

Tubular handling of bicarbonate in dogs with experimental renal failure

In this study, micropuncture experiments were performed to examine the segmental reabsorption of bicarbonate and chloride in the normal dog kidney (stage 1) and in the remnant kidney (stage 3) of dogs with experimental renal failure. The protocol consisted of three phases: (1) 3% extracellular fluid volume expansion, (2) infusion of Ringer's solution containing 90 mM of sodium bicarbonate, and (3) infusion of 150 mM of sodium bicarbonate in Ringer's solution. In the animals with remnant stage 3 kidneys, the ratio of absolute bicarbonate reabsorption/absolute sodium reabsorption was increased compared to dogs with stage 1 kidneys at both the proximal and distal sampling sites. These data suggest that bicarbonate reabsorption was elevated in both the distal and the proximal tubules in experimental chronic renal failure. In contrast to the reabsorption of bicarbonate, chloride reabsorption was depressed in stage 3 kidneys at the late proximal puncture site. This resulted in greater delivery to the distal nephron. The distal segments reabsorbed a constant fraction of delivered chloride, resulting in an increase in fractional chloride excretion in chronic renal failure compared to that in normal animals.

Glutamine transport in renal basolateral vesicles from dogs with metabolic acidosis.

To determine whether the increased ammonia production per nephron in chronic metabolic acidosis is accompanied by augmented L-glutamine transport across the basolateral membrane of the renal cortical cell and consequent increased availability of this ammoniagenic amino acid, we measured L-[3H]glutamine transport in basolateral membrane vesicles (BLMV) isolated from kidneys of normal and acidotic dogs. Na+ -dependent electrogenic transport of L-[3H]glutamine was demonstrated in BLMV from kidneys of normal dogs that exhibited saturability over the concentration range of 25 microM to 2 mM L-glutamine. The apparent Km was 416 +/- 114 microM and Vmax was 536 +/- 129 pmol X mg protein-1 X 15 s-1. The initial rate of Na+ -dependent L-[3H]glutamine transport was increased in BLMV from kidneys of acidotic dogs, as reflected by an increased apparent Vmax. We conclude that an adaptation resulting in greater uptake of L-glutamine across the basolateral membrane of the renal cortical cell may underlie, in part, the increased rate of ammonia production per nephron seen in chronic metabolic acidosis.

Zonal heterogeneity of prostaglandin and thromboxane release in the dog kidney

The release of prostaglandin(PG) and thromboxane(TX) was examined in the six different areas of the normal dog kidney, i.e., renal arterial and venous strips(RA and RV), superficial and deep cortical slices (SC and DC) and outer and inner medullary slices(Om and IM). These tissues were incubated in Krebs-bicarbonate buffer(pH 7.4, 37°C), and the released PGE2, PGF2α, 6-keto-PGF1α and TXB2(as stable metabolites of PG12 and TXA2, respectively) were determined by radioimmunoassay. In RA, RV, SC and DC, 6-keto-PGF1α was predominant, however, there were no quantitative differences between RA and RV, or SC and DC. The release of 6-keto-PGF1α reached a maximum in IM, similar to findings on the release of PGE2 and PGF2α. The release of TXB was uniform in OM and IM. The amount of PGE2, PGF2α, 6-keto-PGF1α and TXB2 released from IM was 2800, 400, 60 and 50 times higher, respectively, than the extent of the release from the cortical slices. These results suggest that PG12 as well as PGE2 and PGF2α, may be involved in renal PG, and that TXA2 is biosynthesized in the normal dog kidney.

Divalent ion transport in dogs with experimental chronic renal failure.

Micropuncture studies were performed to examine the segmental reabsorption of sodium, calcium, and magnesium in the normal dog kidney (stage I) and in the remnant kidney both in the presence (stage II) and in the absence (stage III) of the contralateral normal kidney. The protocol consisted of an initial phase of hydropenia, followed by 5% extracellular fluid volume expansion in the second phase, followed by parathyroid hormone administration in the final phase. In stage II dogs during hydropenia, proximal and distal transport of sodium, calcium, and magnesium were similar to those of normal dogs (stage I). Following 5% body weight volume expansion, fractional deliveries to both the proximal and distal puncture sites were increased similarly in stage 1 and stage II, with a slightly greater increase in stage II animals. In stage III dogs, proximal fractional reabsorption was depressed, as reflected by a marked reduction in proximal tubule fluid to plasma inulin ratios during hydropenia, and the response to volume expansion was accentuated. In the loop segment a constant fraction of the augmented load of sodium, calcium, and magnesium was reabsorbed in stage III. The percentage of the delivered load that was reabsorbed by this segment was similar in all three stages. The diminution in proximal reabsorption in stage III resulted in greater delivery to the distal nephron. The distal reabsorption of a constant fraction of delivered solute resulted in an increase in fractional urinary excretion of sodium, calcium, and magnesium in stage III. Parathyroid hormone significantly reduced the renal excretion of calcium and magnesium in the stage III dogs, indicating the preservation of the renal response to parathyroid hormone in azotemia.

Increased Na+-H+ exchange in brush border vesicles from dogs with renal failure.

In the remnant kidney model of chronic renal failure, absolute reabsorption of Na+ in the proximal tubule of the remaining nephrons is increased over normal. Absolute proximal tubular reabsorption of bicarbonate and proximal tubular H+ excretion per nephron have also been shown to be increased over normal in his model of renal disease. Na+ uptake in membrane vesicles isolated from the brush border membrane of remnant kidneys of dogs with chronic renal failure is increased over uptake in membrane vesicles isolated from kidneys of normal dogs. In the present studies an amiloride-sensitive, electroneutral Na+-H+ exchanger was identified in canine renal brush border membrane vesicles. Na+ uptake in membrane vesicles in the presence of an initial H+ gradient (intravesicular pH less than extravesicular pH) was increased in membrane vesicles isolated from the remnant kidneys of dogs with chronic renal failure over that in membrane vesicles from kidneys of normal dogs. This increase was abolished by amiloride. It is possible that the alterations in Na+ and bicarbonate reabsorption and H+ excretion in the remnant kidney model of chronic renal failure can be explained on the basis of increased activity of the Na+-H+ exchanger in the renal brush border membrane.