Urine-concentrating Ability Research Articles

Transgenic Restoration of Urea Transporter A1 Confers Maximal Urinary Concentration in the Absence of Urea Transporter A3.

Urea has a critical role in urinary concentration. Mice lacking the inner medullary collecting duct (IMCD) urea transporter A1 (UT-A1) and urea transporter A3 (UT-A3) have very low levels of urea permeability and are unable to concentrate urine. To investigate the role of UT-A1 in the concentration of urine, we transgenically expressed UT-A1 in knockout mice lacking UT-A1 and UT-A3 using a construct with a UT-A1 gene that cannot be spliced to produce UT-A3. This construct was inserted behind the original UT-A promoter to yield a mouse expressing only UT-A1 (UT-A1(+/+)/UT-A3(-/-)). Western blot analysis demonstrated UT-A1 in the inner medulla of UT-A1(+/+)/UT-A3(-/-) and wild-type mice, but not in UT-A1/UT-A3 knockout mice, and an absence of UT-A3 in UT-A1(+/+)/UT-A3(-/-) and UT-A1/UT-A3 knockout mice. Immunohistochemistry in UT-A1(+/+)/UT-A3(-/-) mice also showed negative UT-A3 staining in kidney and other tissues and positive UT-A1 staining only in the IMCD. Urea permeability in isolated perfused IMCDs showed basal permeability in the UT-A1(+/+)/UT-A3(-/-) mice was similar to levels in wild-type mice, but vasopressin stimulation of urea permeability in wild-type mice was significantly greater (100% increase) than in UT-A1(+/+)/UT-A3(-/-) mice (8% increase). Notably, basal urine osmolalities in both wild-type and UT-A1(+/+)/UT-A3(-/-) mice increased upon overnight water restriction. We conclude that transgenic expression of UT-A1 restores basal urea permeability to the level in wild-type mice but does not restore vasopressin-stimulated levels of urea permeability. This information suggests that transgenic expression of UT-A1 alone in mice lacking UT-A1 and UT-A3 is sufficient to restore urine-concentrating ability.

TRPC3 Facilitates Adaptation of Collecting Duct Cells to Changes in Osmolality

Mechanical stress induced by changes in tubular flow rate and osmotic gradients is viewed to be an important regulator of the water and electrolytes transport in the collecting duct (CD). Our recent experimental effort identified that pharmacological inhibition or genetic ablation of TRPV4 precluded [Ca2+]i responses to elevated flow but not to hypotonicity suggesting involvement of discrete signaling determinants. Here we combined direct measurements of [Ca2+]i dynamics in freshly isolated split‐opened CDs with genetic tools to unravel molecular components of cellular responses to decreased osmolality. [Ca2+]i elevations elicited by hypotonicity in CD cells were abolished upon removal of extracellular Ca2+, but not by depletion of the endoplasmic reticulum stores, indicating a critical role of direct Ca2+ entry via the plasma membrane. Inhibition of Ca2+‐permeable TRPC3 channel markedly diminishes the amplitude of [Ca2+]i signal induced by hypotonic stimuli in the CD. Interestingly, genetic deletion of TRPC3 significantly increases the activation threshold for [Ca2+]i responses to changes in extracellular osmolality. Water deprivation (18 hours) greatly augmented sensitivity of CD cells to extracellular osmolality in wild‐type, but only mildly in TRPC3‐/‐ mice. At the systemic level, TRPC3 ablation results in a transient defect in urinary concentration ability in response to water deprivation. Overall, our findings demonstrate that elevated flow and osmotic gradients recruit distinct signaling cascades to increase [Ca2+]i in CD cells. Genetic ablation of TRPC3 disturbs sensitivity of CD cells to hypotonic stress contributing to delayed adaptation of the kidney to restricted water intake.

Adaptations to life on arid islands of South Africa: renal performance and phenotypic plasticity in the European rabbit Oryctolagus cuniculus

Morphological parameters of the mammalian kidney can provide information on the extent to which an animal is able to regulate water relations within its body and cope with arid conditions. Certain groups of mammals, such as rabbits and hares, show a flexible phenotype (phenotypic plasticity) with respect to renal physiology that enables them to inhabit a variety of bioclimatic regions.We investigated kidney morphology in the European rabbit, Oryctolagus cuniculus, located on two isolated islands off the coast of the Western Cape South Africa, at sites with varying water and food availability, to ascertain whether these populations demonstrate differences in renal morphology that would link to differential renal physiology. In summer, Dassen Island is more restricted in available water and plants than Robben Island; we tested the prediction that rabbits from Dassen Island would be physiologically more suited to waterlimited conditions. Our data showed that O. cuniculus exhibits phenotypic plasticity, with Dassen Island rabbits displaying renal adaptations more suited to survival in drier environments, such as an increased relative medullary thickness and a higher predicted maximal urineconcentrating ability. This suggests increased renal performance in the Dassen Island population with the consequent increased ability to concentrate urine and conserve body water.

Long-Term Treatment of Cyclosporine Decreases the Expression of Brain-Derived Neurotrophic Factor in Renal Collecting Duct.

Background:Brain-derived neurotrophic factor (BDNF) is originally expressed in central nervous system, but the expression and role of BDNF in chronic cyclosporine (CsA) nephropathy has not to be illustrated. The present study examined the expression of BDNF and its receptors (tyrosine kinase receptors) on kidney in a rat model of chronic cyclosporine (CsA) nephropathy. Methods: Sprague-Dawley rats kept on a low salt diet (0.05% sodium) were treated the VH (vehicle, olive oil, 1 mL/kg s.c.) or CsA (15 mg/kg s.c.) with/without vasopressin infusion. Urine volume, renal histology, and oxidative stress (8-hydroxy-2'-deoxyguanosine,8-OHdG) were compared for different treatment groups.The expression of BDNF,tyrosine receptor kinase (Trk) B and TrkC was evaluated with immunohistochemistry, immunofluorescence, and immunoblotting. Apoptosis was also measured with terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) in tissue sections. Results:Four weeks treatment of CsA increased urine volume, renal fibrosis, and oxidative stress which were accompanied with the decreased BDNF, TrkB and TrkC. The expression of BDNF and TrkB and TrkC were constitutively localized in the collecting tubules in VH group, which were confirmed by double immunofluorescence with AQP-2, but these were markedly decreased in CsA treatmentin same tubular cells. CsA treatment significantly increased the number of TUNEL-positive cells and this was well correlated with the BDNF expression (r = 0.778, P < 0.01). Vasopressin-infused rat treating CsA showed the restored the urine volume and the expression of BDNF and its receptors also recovered to the normal level. Conclusion: Our observations suggest that long-term treatment of CsA inhibits BDNF and its receptor expression in the renal collecting tubule, and that this may be associated with impairment of urine concentration ability.

Lithium safety and tolerability in mood disorders: a critical review

Background : Lithium is a first-line treatment for bipolar disorder in all phases, also indicated as add-on drug for unipolar depression and suicide prevention. This study encompasses a broad critical review on the safety and tolerability of lithium for mood disorders. Methods : A computerized search for English written human studies was made in MEDLINE, using the keywords “lithium” and “mood disorders”, starting from July 1993 through July 2013 (n = 416). This initial search aimed to select clinical trials, prospective data, and controlled design studies of lithium treatment for mood disorders reporting adverse effects (n = 36). The final selection yielded 91 studies. Results : The most common general side effects in patients on lithium treatment were thirst, frequent urination, dry mouth, weight gain, fatigue and cognitive complaints. Lithium users showed a high prevalence of hypothyroidism, hyperparathyroidism, and decrease in urinary concentration ability. Reduction of glomerular filtration rate in patients using lithium was also observed, but in a lesser extent. The evidence of teratogenicity associated with lithium use is not well established. Anti-inflammatory non-steroidal drugs, thiazide diuretics, angiotensin-converting enzyme inhibitors, and alprazolam may increase serum lithium and the consequent risk for intoxication. Discussion : Short-term lithium treatment is associated with mild side effects. Medium and long-term lithium treatment, however, might have effects on target organs which may be prevented by periodical monitoring. Overall, lithium is still a safe option for the treatment of mood disorders.

Abstract 421: Dehydration Upregulates the Full-length (Pro)renin Receptor Expression in the Kidney of Wistar-Kyoto Rats and Spontaneously Hypertensive Rats

(Pro)renin receptor ((P)RR) was newly identified as a member of the renin-angiotensin system (RAS). (P)RR exists in two forms; full-length form (f(P)RR) and soluble form (s(P)RR). The truncated hydrophilic s(P)RR is generated by furin cleavage in the trans-Golgi network and is secreted into the extracellular space. Compared with Wistar-Kyoto rats (WKY), spontaneously hypertensive rats (SHR) demonstrated low urine concentration ability and resistance to vasopressin in a state of dehydration (DH). RAS system contributes to the regulation of body fluid balance, but roles of (P)RR in a state of dehydration (DH) are not clarified. The purpose of this study was to examine the effects of DH on the (P)RR expression in the kidney of WKY and SHR. Twelve week-old, male WKY and SHR were randomly divided into two groups (n=5 in each group); control and DH groups. DH was induced by water restriction for 72 hours in metabolic cage. After 72 hours, rats were killed, and (P)RR protein levels in plasma, urine and kidney were examined by using Western blot. (P)RR mRNA levels were examined by using competitive RT-PCR methods. In addition, furin protein levels were also investigated. DH increased urine osmolarity in WKY and SHR (4090±451 and 3796±429 mOsm/kg). DH significantly decreased the s(P)RR levels in the plasma of WKY and SHR (0.3- and 0.7-fold, P&lt;0.01), but significantly increased in the urine of these rats (1.5- and 3.3-fold, P&lt;0.05). DH significantly increased the f(P)RR levels in the renal homogenates, microsome fraction, and cytosol fraction of WKY and SHR (WKY: 2.3-, 3.0-, and 22.9-fold, SHR: 2.3-, 2.1-, and 7.9-fold, P&lt;0.01). DH did not change the s(P)RR levels in the renal homogenates and cytosol fraction of these rats. In the kidney, DH did not change (P)RR mRNA levels in WKY, but significantly increased them in SHR (2.7-fold, P&lt;0.05). DH significantly decreased furin protein levels in WKY (0.7-fold, P&lt;0.05), but did not change them in SHR. These results indicate that DH upregulates the f(P)RR expression in the kidney of WKY and SHR. The DH-upregulated f(P)RR expression might be mediated through the decrease of furin levels in WKY and increase of (P)RR mRNA levels in SHR.

The conundrum of protection from AKI by adenosine in rodent clamp ischemia models

Kim et al. show that isoflurane uses a tubule-based transforming growth factor-β/CD73-dependent process that generates adenosine to protect mice from ischemic acute kidney injury (AKI) with effects to prevent the 'no-reflow phenomenon' and decrease inflammation. While direct cytoprotection occurred in culture, extensive research suggests that in vivo adenosine protection from rodent ischemic AKI is mediated by a mutually cooperative mechanism involving blood flow, inflammation, and innate immunity through multiple adenosine receptors with promiscuous actions on diverse cell types.

Pre- or post-treatment with aminoguanidine attenuates a renal distal acidification defect induced by acute ureteral obstruction in rats

Acute unilateral ureteral obstruction (UUO) impairs distal nephron acid secretion and stimulates expression of inducible nitric oxide synthase (iNOS) in post-obstructed kidney (POK). This study investigated the influence of pre- or post-treatment with aminoguanidine as a selective iNOS inhibitor on UUO-induced renal functional disturbances. To induce acute UUO, the left ureter in rats was ligated and released after 24 h. Then, a 3 h clearance period followed by bicarbonate loading and thereafter a 30 min clearance period were allocated. Aminoguanidine was administered either prior to the UUO induction or after release of the obstruction in the different rat groups, while untreated and sham groups received normal saline. During the first clearance period, fractional bicarbonate excretion and urinary pH increased markedly in the POK of the untreated group compared with the left kidney of sham group, and a large drop in the difference between urine and blood pCO2 (U-B pCO2) was observed after bicarbonate loading; all of these parameters were ameliorated in the pre-treated and post-treated groups. However, the UUO-induced decreases in creatinine clearance, sodium reabsorption, urine osmolality, and free-water reabsorption in the POK were attenuated only in the post-treated group. Therefore, the in vivo application of a selective iNOS inhibitor partially improved the acute UUO-induced distal nephron acidification defect, while post-treatment but not pre-treatment with aminoguanidine ameliorated decrements of glomerular filtration, sodium reabsorption, and urine-concentrating ability.

Type of uromodulin mutation and allelic status influence onset and severity of uromodulin-associated kidney disease in mice

Uromodulin-associated kidney disease (UAKD) is a dominant heritable renal disease in humans which is caused by mutations in the uromodulin (UMOD) gene and characterized by heterogeneous clinical appearance. To get insights into possible causes of this heterogeneity of UAKD, we describe the new mutant mouse line Umod(C93F), leading to disruption of a putative disulfide bond which is also absent in a known human UMOD mutation, and compare the phenotype of this new mouse line with the recently published mouse line Umod(A227T). In both mutant mouse lines, which were both bred on the C3H background, the Umod mutations cause a gain-of-toxic function due to a maturation defect of the mutant uromodulin leading to a dysfunction of thick ascending limb of Henle's loop (TALH) cells of the kidney. Umod mutant mice exhibit increased plasma urea and Cystatin levels, impaired urinary concentration ability, reduced fractional excretion of uric acid and nephropathological alterations including uromodulin retention in TALH cells, interstitial fibrosis and inflammatory cell infiltrations, tubular atrophy and occasional glomerulo- und tubulocystic changes, a phenotype highly similar to UAKD in humans. The maturation defect of mutant uromodulin leads to the accumulation of immature uromodulin in the endoplasmic reticulum (ER) and to ER hyperplasia. Further, this study was able to demonstrate for the first time in vivo that the severity of the uromodulin maturation defect as well as onset and speed of progression of renal dysfunction and morphological alterations are strongly dependent on the particular Umod mutation itself and the zygosity status.

An early urea-selective urine-concentrating defect in ADPKD

A defect in urine-concentrating ability is discernable in patients with autosomal dominant polycystic kidney disease (ADPKD) before any decline in glomerular filtration rate, and is associated with elevated concentrations of vasopressin and copeptin. This defect is 'urea-selective' and accordingly, the urine-to-plasma ratio of urea concentration could be a clinically relevant early marker of renal dysfunction in patients with ADPKD.

Sepsis-induced urinary concentration defect is related to nitric oxide–dependent inactivation of TonEBP/NFAT5, which downregulates renal medullary solute transport proteins and aquaporin-2*

Acute kidney injury associated with reduced urinary concentration is a frequent and severe complication during sepsis. The present study addressed the effect of endotoxemia on the functional and molecular mechanisms that determine urinary concentrating ability. Efficient urinary concentration depends on, amongst other factors, the expression of the Cl channel kidney-specific chloride channel 1 and its subunit Barttin, the urea transporter-A1, and the water channel aquaporin 2, all of which are regulated by the transcription factor TonEBP/NFAT5. Experimental animal and cell culture model. University laboratory. Wistar rats and Madin-Darby canine kidney cells. Rats were injected with lipopolysaccharide (5 mg/kg bodyweight intraperitoneal) or vehicle (phosphate-buffered saline) as control. After 24 hrs, urine, blood, and tissue samples from various kidney zones were analyzed for parameters that determine urinary concentration ability. Madin-Darby canine kidney cells were treated under isotonic or hypertonic conditions with the nitric oxide donor S-nitroso-N-acetylpenicillamine. In rats injected with lipopolysaccharide, urine osmolality was reduced by ~40%, along with medullary induction of inducible nitric oxide synthase and a dramatic increase in urinary nitric oxide degradation products nitrite/nitrate. Concomitantly, expressions of ClC-K1, Barttin, urea transporter-A1, and aquaporin 2 were significantly lower. This was associated with the appearance of S-nitrosylated TonEBP/NFAT5, as monitored by the biotin-switch assay and immunoprecipitation, and reduced TonEBP/NFAT5 DNA binding activity in the renal inner medulla. These results were confirmed in Madin-Darby canine kidney cells transfected with a reporter construct driven by the urea transporter-A promoter, in which the nitric oxide donor S-nitroso-N-acetylpenicillamine reduces urea transporter-A reporter activity under isotonic and hypertonic conditions. The present data demonstrate that lipopolysaccharide increases medullary nitric oxide production by iNOS induction, resulting in impairment of the transcriptional activity of TonEBP/NFAT5 by S-nitrosylation. The consequence thereof is reduced expression of TonEBP/NFAT5 target genes ClC-K1, Barttin, urea transporter-A1, and aquaporin 2 that are required for urinary concentration. Our findings may provide further insight into the molecular mechanisms underlying the urinary concentration defect in sepsis.

Lithium and glycogen synthase kinase‐3 inhibition induces lysosomal degradation of aquaporin‐2

Lithium (Li), used as treatment of bipolar disorders can lead to diabetes insipidus which shows decreased expression of aquaporin‐ 2 (AQP2) and polyuria. Li also reduces glycogen synthase kinase‐3 (GSK3) activity. GSK3beta deficient mice show decreased urinary concentration ability. The underlying cellular and molecular mechanisms are not fully understood. We used primary cultured inner medullary collecting duct (IMCD) cells, treated with Li or the GSK3 inhibitor SB216763. IMCD cells endogenously express AQP2 and other principal cell markers. AQP2 expression was verified by western blot analysis and realtime PCR. Treatment for 24h with Li or SB216763 reduced expression by 75%. Realtime PCR showed that Li treatment reduced AQP2 expression while SB216763 had no effect. The proteasom inhibitor MG132 or the lysosomal inhibitor bafilomycin (BF) reduced AQP2 mRNA but not protein expression. Li plus MG132 still reduced AQP2 expression. Li plus BF showed normal AQP2 expression. Furthermore in BF but not in MG132 treated cells AQP2 accumulated in vesicular structures. In conclusion we show that Li and GSK3 inhibition reduced AQP2 protein but only Li also mRNA expression. The accumulation of AQP2 after BF treatment indicates recycling of AQP2 in lysosoms. Li and GSK3 inhibition affect this lysosomal recycling thereby promoting lysosomal degradation of AQP2.

Cell culture models and animal models for studying the patho-physiological role of renal aquaporins.

Aquaporins (AQPs) are key players regulating urinary-concentrating ability. To date, eight aquaporins have been characterized and localized along the nephron, namely, AQP1 located in the proximal tubule, thin descending limb of Henle, and vasa recta; AQP2, AQP3 and AQP4 in collecting duct principal cells; AQP5 in intercalated cell type B; AQP6 in intercalated cells type A in the papilla; AQP7, AQP8 and AQP11 in the proximal tubule. AQP2, whose expression and cellular distribution is dependent on vasopressin stimulation, is involved in hereditary and acquired diseases affecting urine-concentrating mechanisms. Due to the lack of selective aquaporin inhibitors, the patho-physiological role of renal aquaporins has not yet been completely clarified, and despite extensive studies, several questions remain unanswered. Until the recent and large-scale development of genetic manipulation technology, which has led to the generation of transgenic mice models, our knowledge on renal aquaporin regulation was mainly based on in vitro studies with suitable renal cell models. Transgenic and knockout technology approaches are providing pivotal information on the role of aquaporins in health and disease. The main goal of this review is to update and summarize what we can learn from cell and animal models that will shed more light on our understanding of aquaporin-dependent renal water regulation.

The Effect of Enalapril on Renal Resistive Index, Urine Electrolyte Levels and TGF-β1 Levels of Kidney Tissues in Rats with Unilateral Partial Ureteropelvic Junction Obstruction

This study aimed to investigate renal arterial resistive index measurements and urine electrolytes before and after enalapril therapy in a rat model of unilateral ureteropelvic obstruction. The transforming growth factor (TGF)-β1 response of the renal tissue was also investigated. 30 Wistar albino rats were randomly allocated into 5 groups (n=6). Group C rats served as controls. Group S rats had only laparotomy. Group E rats were only treated with enalapril. Rats in group UP and group UPE underwent laparotomy to create a left unilateral moderate partial obstruction. 2 weeks after establishing partial ureteropelvic junction obstruction, group UPE rats were treated with enalapril. Urine was collected over 24 h in all groups. Intrarenal arterial resistive index measurements were performed before and 2 weeks after surgery and after enalapril treatment in group UPE, and before and after enalapril treatment in group E. Rats were sacrificed by intracardiac puncture and left kidneys were harvested to evaluate levels of mRNA TGF-β1. There was no significant difference in ARI values in group E. In group UPE, the difference between ARI values before and after surgery was statistically significant; the difference between ARI values after surgery and after enalapril treatment was also statistically significant. There was no statistically significant intra-group difference in urine electrolyte levels for UP group or UPE group. There was no difference in renal mRNA TGF-β1 levels. Enalapril maintained renal blood flow by decreasing the arterial resistive index and maintained renal tubular function by protecting urine concentration and dilution ability in a rat model with unilateral ureteropelvic junction obstruction.

Urine-concentrating defects exacerbate with age in male offspring with a low-nephron endowment

Fetal uninephrectomy (uni-x) in male sheep at 100 days of gestation (term = 150 days) reduces overall nephron endowment without affecting birth weight. Offspring have a lower glomerular filtration rate (GFR) and elevated mean arterial pressure (MAP) at 6 mo of age. This study investigated whether this reduction in renal function was associated with impaired urine-concentrating ability at 6 mo of age and exacerbated with ageing (4 yr) and examined response to 1) nonpressor dose of exogenous arginine vasopressin (AVP; 0.2 μg·kg(-1)·h(-1) iv) and 2) 30 h of water deprivation. Basal MAP was higher in uni-x animals at both ages, and became further elevated with age compared with the sham group (elevation in MAP with age; sham: ~4 mmHg, uni-x: 9 mmHg, P(group × age) < 0.01). GFR declined with ageing in both groups with the decrease being greater with age in the uni-x group (further 26%, P(group × age) < 0.001). In response to AVP infusion, urine osmolality increased in both treatment groups; this response was significantly lower in the uni-x animals and became further reduced with ageing. Uni-x animals had reduced renal expression of vasopressin-2 receptor and aquaporin-2 at both ages (P < 0.01). The increase in plasma AVP levels in response to dehydration was similar between the treatment groups, suggesting the urine-concentrating defect was associated with these renal gene changes rather than defects in AVP secretion. Renal insufficiency due to a low-nephron endowment increases the risk of hypertension and chronic renal disease and may incur greater vulnerability to physiological challenges such as water deprivation as observed in the uni-x animals.

Wnt signaling and renal medulla formation

The renal medulla, the inner compartment of the metanephric kidney, plays vital roles in the regulation of body water, electrolyte homeostasis, and systemic blood pressure. It is composed of the loops-of-Henle, the medullary collecting ducts, the vasa recta, and the medullary interstitium. Its epithelial and endothelial components display ordered spatial organization. This organization serves as the structural basis for its function in urine concentration. The urine concentration ability of a renal medulla is also related to its length among species. In this review, the current understanding of the molecular and cellular mechanisms underlying renal medulla formation (elongation) is summarized, with a focus on the role of Wnt signaling in this developmental process. Renal medulla blunting and effacement is a common symptom of many renal and urological destructions. The knowledge in renal medulla formation should assist efforts in repair and regeneration of a damaged renal medulla, so to improve renal physiology in diseased situations.

Fluvastatin modulates renal water reabsorption in vivo through increased AQP2 availability at the apical plasma membrane of collecting duct cells

X-linked nephrogenic diabetes insipidus (XNDI), a severe pathological condition characterized by greatly impaired urine-concentrating ability of the kidney, is caused by inactivating mutations in the V2 vasopressin receptor (V2R) gene. The lack of functional V2Rs prevents vasopressin-induced shuttling of aquaporin-2 (AQP2) water channels to the apical plasma membrane of kidney collecting duct principal cells, thus promoting water reabsorption from urine to the interstitium. At present, no specific pharmacological therapy exists for the treatment of XNDI. We have previously reported that the cholesterol-lowering drug lovastatin increases AQP2 membrane expression in renal cells in vitro. Here we report the novel finding that fluvastatin, another member of the statins family, greatly increases kidney water reabsorption in vivo in mice in a vasopressin-independent fashion. Consistent with this observation, fluvastatin is able to increase AQP2 membrane expression in the collecting duct of treated mice. Additional in vivo and in vitro experiments indicate that these effects of fluvastatin are most likely caused by fluvastatin-dependent changes in the prenylation status of key proteins regulating AQP2 trafficking in collecting duct cells. We identified members of the Rho and Rab families of proteins as possible candidates whose reduced prenylation might result in the accumulation of AQP2 at the plasma membrane. In conclusion, these results strongly suggest that fluvastatin, or other drugs of the statin family, may prove useful in the therapy of XNDI.

L‐carnitine improves oxidative stress and suppressed energy metabolism but not renal dysfunction following release of acute unilateral ureteral obstruction in rat

We recently showed that L-carnitine reduced oxidative stress and suppressed energy metabolism, while α-tocopherol only prevented redox imbalance, in the obstructed kidney of rats subjected to 24-hr of unilateral ureteral obstruction (UUO). The present study was undertaken to investigate the effects of both compounds on disturbed renal hemodynamics, solutes-excretion, and urine-concentrating ability as well as renal oxidative stress and suppressed metabolism at early hours following release of 24-hr UUO. UUO was induced in anaesthetized rats that received L-carnitine, α-tocopherol or their vehicles in four different groups. Each rat was re-anaesthetized, prepared for renal functional measurements, and its ureteral obstruction was released at 24-hr. Then, urines of both kidneys were separately collected during 30-90 min of post-release, with taking blood samples at beginning and end of it. Finally, both kidneys were removed and preserved at -80°C for future measuring their levels of ATP and ADP as well as malondialdehyde (MDA) and ferric reducing/antioxidant power (FRAP). There were also sham and control groups. Post-obstructed kidney (POK) of vehicle-treated groups compared to equivalent kidney of sham group had lower ATP, ATP/ADP, FRAP, creatinine clearance, absolute Na(+)- and K(+)-excretion, and effective free-water reabsorption, but higher MDA and ADP. L-carnitine could improve oxidative stress and suppressed energy metabolism and α-tocopherol normalized redox state, but both compounds did not have any effects on altered functional variables of the POK. Oxidative stress and suppressed energy metabolism may not be involved in the development of renal dysfunction during acute ureteral obstruction.

Conditional ablation of glycogen synthase kinase 3β in postnatal mouse kidney

Glycogen synthase kinase (GSK)3 is a ubiquitously expressed serine/threonine kinase existing in two isoforms, namely GSK3α and GSK3β. Aside from the long-recognized role in insulin signal transduction and glycogen biosynthesis, GSK3β has been recently coined as a master control molecule in nuclear factor-κB activation and inflammatory kidney injury. Nevertheless, previous studies are less conclusive because they relied greatly on small molecule inhibitors, which lack selectivity and barely distinguish between the GSK3 isoforms. In addition, early embryonic lethality after global knockout of GSK3β precludes interrogation of the biological role of GSK3β in the adult kidney. To circumvent these issues, the Cre/loxP system was used to generate a conditional knockout mouse model in which the GSK3β gene was specifically deleted in kidney cortical tubules at postnatal mature stage. Kidney-specific ablation of GSK3β resulted in a phenotype no different from control littermates. Knockout mice (KO) were viable and exhibited normal development and normal kidney physiology in terms of kidney function, urine albumin excretion, and urine-concentrating ability. It is noteworthy that apart from normal glomerular and tubulointerstitial morphology, the kidneys from KO demonstrated more glycogen accumulation in the renal cortical tubules as assessed by both periodic acid-Schiff staining for light microscopy and direct biochemical assay, consistent with an elevated glycogen synthetic activity as evidenced by diminished inhibitory phosphorylation of glycogen synthase that occurred subsequent to GSK3β ablation. This finding was further validated by electron microscopic observations of increased deposition of glycogen particles in the renal tubules of KO, suggesting that GSK3α could not fully compensate for the loss of GSK3β in regulating glycogen metabolism in the kidney. Collectively, our study suggests that kidney-specific ablation of GSK3β barely affects kidney function and histology under normal circumstances. Extended examinations of these KO under diseased conditions are merited to understand the role of GSK3β in renal pathophysiology.

Aquaporin-2 urinary excretion in preterm infants: relationship to diuresis and vasopressin

Few investigations have explored the urinary aquaporin-2 (u-AQP2) excretion pattern after birth in preterm infants with conflicting results regarding the correlation between u-AQP2, urinary osmolality and vasopressin. The aims of this study were to evaluate u-AQP2 excretion during the first week of life in preterm infants, to correlate u-AQP2 with other markers of renal function and to investigate the relationship between u-AQP2, urinary tonicity and arginine-vasopressin in the immature kidney. In infants born less than 33 weeks daily diuresis, u-AQP2, urinary arginine-vasopressin, urine and plasma tonicity, creatinine and electrolytes were measured through the first 7 days of life. Fifty-five infants were evaluated. u-AQP2 excretion showed the following profile: the highest u-AQP2 levels were found on day 2 and values remained significantly higher until day 5 with respect to day 1. On day 6, u-AQP2 levels significantly decreased to values closer to those found on day 1. u-AQP2 excretion was not associated with arginine-vasopressin while significant, but weak association was found with urinary tonicity (r = -0.20; -0.32 < r < -0.11; P < 0.05). u-AQP2 excretion and creatinine clearance were significantly associated during the study period (r = 0.19; 0.08 < r < 0.29; P < 0.05). There was a strong association between totally u-AQP2 excretion and diuresis over the week (r = 0.72; 0.66 < r < 0.76; P < 0.0001). Significant variations occur in AQP2 expression levels during the first week of life in preterm infants. AQP2 does not seem to contribute to the urinary concentration ability after birth. Further investigations are required to elucidate the mechanisms underlying the strong association between diuresis and u-AQP2 excretion in early postnatal life.