Urea Reabsorption Research Articles

Identification and Expression of a Putative Facilitative Urea Transporter in Three Species of True Frogs (Ranidae): Implications for Terrestrial Adaptation

Urea transporters (UTs) help mediate the transmembrane movement of urea and therefore are likely important in amphibian osmoregulation. Although UTs contribute to urea reabsorption in anuran excretory organs, little is known about the protein’s distribution and functions in other tissues, and their importance in the evolutionary adaptation of amphibians to their environment remains unclear. To address these questions, we obtained a partial sequence of a putative UT and examined relative abundance of this protein in tissues of the wood frog (Rana sylvatica), leopard frog (R. pipiens), and mink frog (R. septentrionalis), closely related species that are adapted to different habitats. Using immunoblotting techniques, we found the protein to be abundant in the osmoregulatory organs but also present in visceral organs, suggesting that UTs play both osmoregulatory and nonosmoregulatory roles in amphibians. UT abundance seems to relate to the species’ habitat preference, as levels of the protein were higher in the terrestrialR. sylvatica, intermediate in the semiaquaticR. pipiens, and quite low in the aquaticR. septentrionalis. These findings suggest that, in amphibians, UTs are involved in various physiological processes, including solute and water dynamics, and that they have played a role in adaptation to the osmotic challenges of terrestrial environments.

Can the difference in serum concentration of urea and cystatin C be used in diagnosis and prognosis of heart failure?

Changes in renal function are an important diagnostic and prognostic indicator in patients with heart failure (HF). They are caused by decreased renal perfusion and consequently decreased glomerular filtration rate (GFR), or by the effect of increased neurohormonal activity (sympathetic nervous system [SNS], rennin–angiotensin–aldosterone system [RAAS] and arginine vasopressin [AVP]). However, the increase of serum concentration of urea, creatinine and other metabolites is not specific for HF. Therefore, it is not possible to distinguish HF from renal diseases solely based on the increase of their concentration, since the increase of their concentration caused by the decrease of GFR cannot be differentiated from the increase due to neurohormonal activity. Urea and cystatin C (Cys C) have different mechanisms of renal elimination, so it can be assumed that in HF their concentrations will not be increased proportionally, what can be used for diagnostic and prognostic purposes. After glomerular filtration Cy C undergoes proximal tubular reabsorption and breakdown, without returning to the blood flow. Since it is not secreted, its serum concentration depends only on GFR. In contrast to Cys C, urea is filtered in glomerulus and subsequently reabsorbed in proximal tubules and colleting duct. Reabsorption of urea is modified by effects of SNS, RAAS and AVP. Therefore its serum concentration depends upon GFR and neurohormonal effect on the tubular function. Since the increase of serum concentration of Cys C is caused only by the effect of the decreased renal perfusion on GFR, while the increase of urea is a result from both decreased GFR and tubular effects of increased neurohormonal activity, the paper hypothesis is that in HF the increase of urea will be significantly higher than the increase of serum Cys C, while in the patients with renal diseases their increase would be mostly proportional. It can be assumed that the disproportion between the increase of Cys C and urea would indicate an increased neurohormonal activity in patients with HF and correlate with its activity. If this hypothesis is proved correct, this parameter could be used in HF diagnosis and risk stratification of such patients.

Acute Kidney Injury

1. David T. Selewski, MD* 2. Jordan M. Symons, MD† 1. *Department of Pediatrics and Communicable Diseases, Division of Nephrology, C.S. Mott Children’s Hospital, University of Michigan, Ann Arbor, MI. 2. †Department of Pediatrics, Division of Nephrology, University of Washington School of Medicine, Seattle, WA. * Abbreviations: ACE: : angiotensin-converting enzyme AKI: : acute kidney injury AKIN: : Acute Kidney Injury Network ANCA: : antineutrophil cytoplasmic antibody ATN: : acute tubular necrosis ECG: : electrocardiogram FEurea: : fractional excretion of urea FENa: : fractional excretion of sodium KDIGO: : Kidney Disease: Improving Global Outcomes NSAID: : nonsteroidal anti-inflammatory drug RIFLE: : Risk, Injury, Failure, Loss, and End-stage The term acute kidney injury has replaced acute renal failure and represents a spectrum of clinically meaningful kidney damage. After completing this article, readers should be able to: 1. Recognize and define the spectrum of acute kidney injury (AKI). 2. Understand the diagnostic approach and be able to differentiate the main causes of AKI. 3. Understand the complications of AKI and the treatment of a child with AKI. Acute kidney injury (AKI), formerly called acute renal failure, is characterized by multiple abnormalities, including increases in serum creatinine and blood urea nitrogen, electrolyte abnormalities, acidosis, and difficulties with fluid management. We have come to realize that what was previously thought to be relatively minor damage to the kidney can have significant short-term effects on morbidity and mortality and potential long-term implications for the development of chronic kidney disease. Thus, the term acute kidney injury has replaced acute renal failure , suggesting the spectrum of kidney damage that can occur. ### Definition AKI is classically defined as an acute decrease in glomerular filtration rate, which results in an increase in serum creatinine. It is important to recognize the limitations of creatinine as a marker of AKI because an increase in creatinine can be delayed by as much as 48 hours after damage to the kidney has occurred. Despite this limitation, change in creatinine remains the gold standard for the diagnosis of AKI. An evolution in the definition of AKI to better understand, characterize, and study the disease spectrum, has occurred, which has sought to capture the clinical importance of even small variations in kidney function. In addition, previous definitions used in the literature were widely disparate; this …

Acute Kidney Injury

Acute Kidney Injury

An in vitro study of urea, water, ion and CO2/HCO3− transport in the gastrointestinal tract of the dogfish shark (Squalus acanthias): the influence of feeding

In vitro gut sac preparations made from the cardiac stomach (stomach 1), pyloric stomach (stomach 2), intestine (spiral valve) and colon were used to examine the impact of feeding on transport processes in the gastrointestinal tract of the dogfish shark. Preparations were made from animals that were euthanized after 1-2 weeks of fasting, or at 24-48 h after voluntary feeding on a 3% ration of teleost fish (hake). Sacs were incubated under initially symmetrical conditions with dogfish saline on both surfaces. In comparison to an earlier in vivo study, the results confirmed that feeding caused increases in H(+) secretion in both stomach sections, but an increase in Cl(-) secretion only in stomach 2. Na(+) absorption, rather than Na(+) secretion, occurred in both stomach sections after feeding. All sections of the tract absorbed water and the intestine strongly absorbed Na(+) and Cl(-), regardless of feeding condition. The results also confirmed that feeding increased water absorption in the intestine (but not in the colon), and had little influence on the handling of Ca(2+) and Mg(2+), which exhibited negligible absorption across the tract. However, K(+) was secreted in the intestine in both fasted and fed preparations. Increased intestinal water absorption occurred despite net osmolyte secretion into the mucosal saline. The largest changes occurred in urea and CO2/HCO3(-) fluxes. In fasted preparations, urea was absorbed at a low rate in all sections except the intestine, where it was secreted. Instead of an increase in intestinal urea secretion predicted from in vivo data, feeding caused a marked switch to net urea absorption. This intestinal urea transport occurred at a rate comparable to urea reabsorption rates reported at gills and kidney, and was apparently active, establishing a large serosal-to-mucosal concentration gradient. Feeding also greatly increased intestinal CO2/HCO3(-) secretion; if interpreted as HCO3(-) transport, the rates were in the upper range of those reported in marine teleosts. Phloretin (0.25 mmol l(-1), applied mucosally) completely blocked the increases in intestinal urea absorption and CO2/HCO3(-) secretion caused by feeding, but had no effect on Na(+), Cl(-) or water absorption.

Flow stimulates inner medullary collecting duct endothelin‐1 production

Collecting duct (CD)‐derived endothelin‐1 (ET‐1) inhibits Na and water reabsorption; its deficiency causes marked hypertension. CD ET‐1 synthesis is enhanced by extracellular fluid volume expansion. In cultured cortical CD (CCD) cells, increased flow (as would occur in tubule fluid during volume expansion) stimulates ET‐1 production; this is ENaC‐dependent. Since the inner medullary CD (IMCD) is the major site of renal ET‐1 synthesis, we examined the effect of flow on IMCD ET‐1 production. Mouse IMCD3 cells were subjected to static conditions or flow (2 dyne/cm2 for 2 hr), followed by determination of ET‐1 mRNA. Flow increased ET‐1 mRNA by 2.2‐fold. Absence of perfusate Ca prevented the flow response. BAPTA, W‐7, calmidazolium, KN‐93, calphostin C and cyclosporin A reduced the ET‐1 flow response, indicating that PKC and Ca/calmodulin/calmodulin kinase/calcineurin pathways are essential for the flow response. Amiloride or benzamil did not affect the ET‐1 response to flow. Increasing perfusate osmolality to 450 mOsm with NaCl, mannitol or urea elicited a marked flow response (4.4‐fold increase in ET‐1 mRNA). Removal of cilia with chloral hydrate reduced the flow response, while flow failed to stimulate ET‐1 mRNA in mIMCD3 cells deficient in polycystin‐2. These data suggest that IMCD ET‐1 synthesis is stimulated by tubule fluid flow via increased solute delivery and possibly cilia deformation which in turn activates PKC‐ and Ca‐dependent pathways. We propose that Na delivery stimulates CCD ET‐1 which, in turn, inhibits CCD Na reabsorption, while solute and water delivery stimulate IMCD ET‐1 which, in turn, inhibits IMCD water and urea reabsorption.

Abstract 19062: Copeptin-to-BUN Ratio Predicts Outcome in Patients with Advanced Heart Failure: A key to the use of vasopressin antagonists?

Background: The arginine-vasopressin-aldosteron system is activated in patients with chronic heart failure and elevated serum concentrations of vasopressin as well as the c-terminal portion of its prohormone (copeptin) strongly predicts outcome of these patients. Re-absorption of urea by the kidneys is regulated by vasopressin and thereby it also influences blood urea nitrogen (BUN) concentrations. In the present study we investigated, whether the ratio of serum copeptin and BUN (copeptin-to-BUN) could be used to estimate long-term outcome of patients with chronic stable heart failure (CHF). Methods: We measured copeptin, BUN, high-sensitivity troponin T (hs-cTnT), plasma MR-proANP, MR-proADM and Nt-proBNP concentrations in 174 patients with CHF. Patients were followed for all-cause mortality and rehospitalization due to heart failure during a median time of 834days. Results: The ratio of copeptin-to-BUN was significantly higher among patients who died or were re-admitted to the hospital compared to those who remained event-free (0.86 vs. 0.45; p<0.001). In univariate Cox regression analysis, patients with copeptin-to-BUN ratio higher than the median had significantly higher risk of death or hospital re-admission than patients with lower copeptin-to-BUN ratio (HR 2.16 p<0.001; Figure). In multivariate analyses adjusted for several clinical variables and laboratory measures - including Nt-proBNP, MR-proADM, MR-proANP, hs-cTnT - the ratio of copeptin-to-BUN remained a highly significant determinant of long-term outcome (HR 1.645; p=0.023). Conclusion: In addition to well-established modern biomarkers, the copeptin-to-BUN ratio is a significant independent determinant of survival of patients with CHF. Future studies should evaluate whether copeptin-to-BUN ratio could be used to identify patients with advanced heart failure who might benefit from the use of vasopressin antagonists.

Effect of sodium chloride intake on urine volume, urinary urea excretion, and milk urea concentration in lactating dairy cattle

Milk urea nitrogen (MUN; mg of N/dL) has been shown to be related to excretion of urinary urea N (UUN; g of N/d) and total excretion of urinary N (UN; g of N/d) in dairy cows. In the present experiment, it was hypothesized that MUN and the relationship between MUN and UUN or UN is affected by urine volume as a result of dietary sodium chloride intake. Twelve lactating Holstein-Friesian dairy cows (mean ± SD: milk production 28.1±3.23kg/d and 190±41d in milk), of which 4 were fitted with catheters in the urine bladder and jugular vein, were randomly assigned to 4 dietary levels of sodium chloride (3, 9, 14, and 19g of Na/kg of DM) according to a triple 4×4 Latin square design. Cows were fed at 95% of ad libitum intake, excluding salt addition. Milk was analyzed for MUN and protein content; urine was analyzed for total N, urea, and creatinine content; feces were analyzed for total N and DM content; and blood plasma was analyzed for urea and creatinine content. Creatinine clearance rate (CCR; L/min) and renal urea reabsorption ratio were estimated based on plasma concentrations of urea and creatinine, and total excretion of urea and creatinine in urine. Intake of DM and N, milk production, and milk protein content were (mean ± SD), on average, 21.4±1.24kg/d, 522±32.0g/d, 25.4±2.53kg/d, and 3.64±0.186%, respectively. A linear relationship was found between Na intake and urine production [urine (kg/d; mean ± SE)=7.5±4.33+0.136±0.0143 × Na intake (g/d)] and between Na intake and MUN [MUN (mg/dL; mean ± SE)=13.5±0.35−0.0068±0.00104 × Na intake (g/d)]. Despite the decrease in MUN with increased Na intake, UN excretion increased linearly with Na intake. Excretion of UUN was not affected by dietary Na content. A linear plateau relationship was observed between CCR and renal urea reabsorption. An increase in CCR coincided with an increase in calculated renal urea reabsorption until a CCR breakpoint value (mean ± SD) of 1.56±0.063 L/min was reached. We conclude that Na intake is negatively related to MUN, whereas UUN is not affected. Variation in mineral intake levels that affect urine volume should, therefore, be taken into account when using MUN as an indicator of UUN in dairy cattle.

A review of factors influencing milk urea concentration and its relationship with urinary urea excretion in lactating dairy cattle

SUMMARYMilk urea nitrogen (MUN) concentration in dairy cows may serve as an on-farm indicator to guide nutritional strategies and to help reduce emissions of nitrogen (N) to the environment. Excretion of urinary urea nitrogen (UUN) is positively related to MUN, but the relationship is highly variable. The accuracy of MUN as a predictor of UUN may improve when various factors that affect this relationship can be taken into account. The current review discusses the impact of a number of UUN : MUN ratio influencing factors related to: physiological mechanisms in the dairy cow, farm management, differences between individual cows, nutrition and analysis methods for MUN. Factors related to variation in water intake, urine production, dietary protein level, body weight (BW) and time and frequency of feeding and milking are shown to affect MUN and its relationship with UUN. In addition, a number of factors are discussed that are likely to affect this relationship such as biological rhythm, renal reabsorption of urea during periods of protein deficiency and breeding value for MUN. Accounting for these above-mentioned factors in the relationship between MUN and UUN might substantially improve the applicability and accuracy of MUN as a predictor of protein utilization efficiency and UUN.

Expression of urea transporters is affected by dietary nitrogen restriction in goat kidney1

Ruminants are known to be able to very effectively recycle urinary urea and reuse it as a source of N for ruminal microbes. It is presumed that urea recycling is accomplished by specialized urea transporters (UT) which are localized in the kidney. This could be especially important in times of increased N requirement, such as during growth or during reduced dietary N intake. The aim of our study was to characterize and to localize UT in the goat (capra hircus) kidney and to investigate its response to reduced dietary N intake in growing goats. Therefore, 12 growing, male goats were fed either a diet containing high (17% CP in complete diet) or low (9% CP in complete diet) N content for 6 wk. After harvesting, blood and kidney samples were taken and analyzed. The mRNA of the different UT isoforms, UT-A1, UT-A2 and UT-B, were detected semiquantitatively in renal tissue by Northern blot analysis. For UT-A2 and UT-B, no statistically significant effect of dietary N restriction on renal mRNA expression could be detected (UT-A2: P = 0.26, UT-B: P = 0.07). However, renal mRNA abundance of UT-A1 significantly increased in the kidney of low-N-fed goats (P = 0.01). Furthermore, protein amounts of UT-B were verified by western blotting; and the localization of UT-A2 and UT-B protein was demonstrated by immunohistochemistry. No significant differences in protein amounts of UT-B could be observed comparing the 2 feeding groups (P = 0.78). The UT-B was localized in renal medulla and papilla, whereas UT-A2 was only found in renal medulla. In addition, comparison of UT-A and UT-BAA sequences of monogastric animals and ruminants showed a high degree of homology, indicating a similar function of the transporters among these species. In summary, we conclude that in ruminants, urea reabsorption in the kidney is most likely increased in response to a low-N diet via an upregulation of UT-A1 mRNA expression. Hypothetically, the reabsorbed urea can then be returned to the rumen via the bloodstream and thus be reused as a source of N for protein synthesis of ruminal microbial community.

Seasonal Variation and Response to Osmotic Challenge in Urea Transporter Expression in the Dehydration‐ and Freeze‐Tolerant Wood Frog, Rana sylvatica

Urea accumulation is a universal response to osmotic challenge in anuran amphibians, and facilitative urea transporters (UTs) seem to play an important role in this process by acting in the osmoregulatory organs to mediate urea retention. Although UTs have been implicated in urea reabsorption in anurans, little is known about the physiological regulation of UT protein abundance. We examined seasonal variation in and effects of osmotic challenge on UT protein and mRNA levels in kidney and urinary bladder of the wood frog (Rana sylvatica), a terrestrial species that tolerates both dehydration and tissue freezing. Using immunoblotting techniques to measure relative UT abundance, we found that UT numbers varied seasonally, with a low abundance prevailing in the fall and winter, and higher levels occurring in the spring. Experimental dehydration of frogs increased UT protein abundance in the urinary bladder, whereas experimental urea loading decreased the abundance of UTs in kidney and bladder. Experimental freezing, whether or not followed by thawing, had no effect on UT numbers. UT mRNA levels, assessed using quantitative real-time polymerase chain reaction, did not change seasonally nor in response to any of our experimental treatments. These findings suggest that regulation of UTs depends on the nature and severity of the osmotic stress and apparently occurs posttranscriptionally in response to multiple physiological factors. Additionally, UTs seem to be regulated to meet the physiological need to accumulate urea, with UT numbers increasing to facilitate urea reabsorption and decreasing to prevent retention of excess urea.

The sweet side of urea transporters

UREA IS PRODUCED AS AN END product of protein catabolism in the liver and is ultimately excreted by the kidney. However, this unique small molecule of only 60 daltons is also used by the kidney to control water balance by a complex process that involves both secretion and reabsorption of urea throughout the entire renal tubule (11). Disturbance of these processes can cause a toxic buildup of urea in the blood termed uremia. Clearly, urea transporters (UT) are key players in this important process, and understanding UT activity, synthesis, turnover, and membrane trafficking has been a focus of researchers since the first gene was cloned in 1993 (17).

Glycoforms of UT-A3 urea transporter with poly-N-acetyllactosamine glycosylation have enhanced transport activity

Urea transporters UT-A1 and UT-A3 are both expressed in the kidney inner medulla. However, the function of UT-A3 remains unclear. Here, we found that UT-A3, which comprises only the NH(2)-terminal half of UT-A1, has a higher urea transport activity than UT-A1 in the oocyte and that this difference was associated with differences in N-glycosylation. Heterologously expressed UT-A3 is fully glycosylated with two glycoforms of 65 and 45 kDa. By contrast, UT-A1 expressed in HEK293 cells and oocytes exhibits only a 97-kDa glycosylation form. We further found that N-glycans of UT-A3 contain a large amount of poly-N-acetyllactosamine. This highly glycosylated UT-A3 is more stable and is enriched in lipid raft domains on the cell membrane. Kifunensine, an inhibitor of α-mannosidase that inhibits N-glycan processing beyond high-mannose-type N-glycans, significantly reduced UT-A3 urea transport activity. We then examined the native UT-A1 and UT-A3 glycosylation states from kidney inner medulla and found the ratio of 65 to 45 kDa in UT-A3 is higher than that of 117 to 97 kDa in UT-A1. The highly stable expression of highly glycosylated UT-A3 on the cell membrane in kidney inner medulla suggests that UT-A3 may have an important function in urea reabsorption.

Hydration status affects urea transport across rat urothelia

Although mammalian urinary tract epithelium (urothelium) is generally considered impermeable to water and solutes, recent data suggest that urine constituents may be reabsorbed during urinary tract transit and storage. To study water and solute transport across the urothelium in an in vivo rat model, we instilled urine (obtained during various rat hydration conditions) into isolated in situ rat bladders and, after a 1-h dwell, retrieved the urine and measured the differences in urine volume and concentration and total quantity of urine urea nitrogen and creatinine between instilled and retrieved urine in rat groups differing by hydration status. Although urine volume did not change >1.9% in any group, concentration (and quantity) of urine urea nitrogen in retrieved urine fell significantly (indicating reabsorption of urea across bladder urothelia), by a mean of 18% (489 mg/dl, from an instilled 2,658 mg/dl) in rats receiving ad libitum water and by a mean of 39% (2,544 mg/dl, from an instilled 6,204 mg/dl) in water-deprived rats, but did not change (an increase of 15 mg/dl, P = not significant, from an instilled 300 mg/dl) in a water-loaded rat group. Two separate factors affected urea nitrogen reabsorption rates, a urinary factor related to hydration status, likely the concentration of urea nitrogen in the instilled urine, and a bladder factor(s), also dependent on the animal's state of hydration. Urine creatinine was also absorbed during the bladder dwell, and hydration group effects on the concentration and quantity of creatinine reabsorbed were qualitatively similar to the hydration group effect on urea transport. These findings support the notion(s) that urinary constituents may undergo transport across urinary tract epithelia, that such transport may be physiologically regulated, and that urine is modified during transit and storage through the urinary tract.

Spontaneous bladder perforation: a rare complication of tuberculosis

Spontaneous bladder perforation secondary to tuberculosis (TB) is very rare. Only three cases have been reported so far in the literature. Due to its rarity, the diagnosis of spontaneous bladder perforation is often missed. Confirmation of TB via culture takes a long time and starting empirical treatment for TB is necessary. We relate our experience with a young woman who presented with clinical features of a perforated appendix and was only diagnosed with bladder perforation during laparotomy. She also had distal right ureteral stricture and left infundibular stenosis. The provisional diagnosis of TB was attained via typical histopathological features and a positive Mantoux test. She was started empirically on anti-TB treatment and recovered without any complications. Urine culture after 6 weeks confirmed the diagnosis of TB.

Subcellular distribution of urea transporter in the collecting tubule of shark kidney is dependent on environmental salinity

In the kidney of marine elasmobranchs, urea reabsorption from filtered urine is essential for maintaining high levels of urea in the body. In the kidney of the houndshark, Triakis scyllium, we previously found that a facilitative urea transporter (UT) is localized to a specific nephron segment, the collecting tubule, suggesting that the collecting tubule has an important role in the urea reabsorption process. To elucidate the roles of UT, we transferred T. scyllium to high (130%) and low (30%) salinity, and examined UT mRNA levels and UT distribution patterns in the kidney using real-time PCR and semi-quantitative fluorescence immunohistochemistry, respectively. Following transfer to low and high salinity, houndshark decreased and increased plasma urea concentrations, respectively, in order to control plasma osmolality. The abundance of UT mRNA did not differ among the experimental groups, whereas that of UT protein in the collecting tubule was significantly decreased in 30% seawater (SW). Furthermore, the subcellular UT distribution was dramatically changed. UT in the apical plasma membrane of collecting tubule almost disappeared in 30% SW, whereas it slightly increased in 130% SW compared with 100% SW. Conversely, reverse transfer of fish from 30 to100% SW restored UT in the apical membrane. These results indicate that the accumulation of UT to the apical plasma membrane of the collecting tubule of Triakis is an important factor for regulating urea reabsorption in the kidney.

Multiple urea transporter proteins in the kidney of holocephalan elephant fish (Callorhinchus milii)

Reabsorption of filtered urea by the kidney is essential for retaining high levels of urea in marine cartilaginous fish. Our previous studies on the shark facilitative urea transporter (UT) suggest that additional UT(s) comprising the urea reabsorption system could exist in the cartilaginous fish kidney. Here, we isolated three cDNAs encoding UTs from the kidney of elephant fish, Callorhinchus milii, and termed them efUT-1, efUT-2 and efUT-3. efUT-1 is orthologous to known elasmobranch UTs, while efUT-2 and efUT-3 are novel UTs in cartilaginous fish. Two variants were found for efUT-1 and efUT-2, in which the NH(2)-terminal intracellular domain was distinct between the variants. Differences in potential phosphorylation sites were found in the variant-specific NH(2)-terminal domains. When expressed in Xenopus oocytes, all five UT transcripts including the efUT-1 and efUT-2 variants induced more than a 10-fold increase in [(14)C] urea uptake. Phloretin inhibited dose-dependently the increase of urea uptake, suggesting that the identified UTs are facilitative UTs. Molecular phylogenetic analysis revealed that efUT-1 and efUT-2 had diverged in the cartilaginous fish lineage, while efUT-3 is distinct from efUT-1 and efUT-2. The present finding of multiple UTs in elephant fish provides a key to understanding the molecular mechanisms of urea reabsorption system in the cartilaginous fish kidney.

Why does the plasma urea concentration increase in acute dehydration?

Acute dehydration biochemically manifests itself with mild uremia and a normal creatinine concentration. The reason for this, as quoted in popular medical textbooks, is that there is increased reabsorption of urea by the kidneys ([1][1]). This explanation is rarely taken further by clinical tutors

MDM2 E3 ubiquitin ligase mediates UT-A1 urea transporter ubiquitination and degradation

UT-A1 is the primary urea transporter in the apical plasma membrane responsible for urea reabsorption in the inner medullary collecting duct. Although the physiological function of UT-A1 has been well established, the molecular mechanisms that regulate its activity are less well understood. Analysis of the UT-A1 amino acid sequence revealed a potential MDM2 E3 ubiquitin ligase-binding motif in the large intracellular loop of UT-A1, suggesting that UT-A1 urea transporter protein may be regulated by the ubiquitin-proteasome pathway. Here, we report that UT-A1 is ubiquitinated and degraded by the proteasome but not the lysosome proteolytic pathway. Inhibition of proteasome activity causes UT-A1 cell surface accumulation and concomitantly increases urea transport activity. UT-A1 interacts directly with MDM2; the binding site is located in the NH2-terminal p53-binding region of MDM2. MDM2 mediates UT-A1 ubiquitination both in vivo and in vitro. Overexpression of MDM2 promotes UT-A1 degradation. The mechanism is likely to be physiologically important as UT-A1 ubiquitination was identified in kidney inner medullary tissue. The ubiquitin-proteasome degradation pathway provides an important novel mechanism for UT-A1 regulation.

Mechanisms to concentrate the urine: an opinion

Our goal is to suggest how the renal concentrating mechanism is regulated in vivo. The majority of descending thin limbs of the loop of Henle lack aquaporin-1 water channels, and loops of Henle in the inner medulla lack urea transporters. Lack of water permeability in the descending thin limbs of the loop of Henle offers several advantages. First, since much less water is added to the outer medullary interstitial compartment, inhibitory control mechanisms on sodium and chloride reabsorption from the medullary thick ascending of loop of Henle initiated by water addition from the medullary collecting duct can be effective. Second, recycling of urea is efficient, as little urea will be washed out of the medulla. Third, delivery of a larger volume of filtrate to the medullary thick ascending limb of the loop of Henle permits both an appreciable reabsorption of sodium along with only a small fall in the luminal concentration of sodium in each of these liters. Hence there need be only a small lumen positive voltage in the medullary thick ascending limb of the loop of Henle. The absence of urea transporters in the loop of Henle in the inner medulla is required for a passive mechanism of sodium and chloride reabsorption in the inner medulla. Control of urea reabsorption from the medullary collecting duct is needed to prevent excessive oliguria in electrolyte-poor urine.