Biomedical subjects

O Levillain

Publications and source records attributed to O Levillain.

At least 19 recordsLinked to original sources

PubMed2001

Gene expression of the taurine transporter and taurine biosynthetic enzymes in rat kidney after antidiuresis and salt loading.

Taurine is thought to be an osmolyte in the kidney medulla. We have investigated the gene expression of the taurine transporter (TauT) and the enzymes of taurine biosynthesis, cysteine dioxygenase (CDO) and cysteine sulfinate decarboxylase (CSD). We achieved this by measuring their mRNA levels using reverse transcriptase polymerase chain reaction (RT-PCR) in five kidney regions of rats in various hydration states; namely, normal hydration, after 2 days of antidiuresis following chronic diuresis and finally after acute salt loading. The mRNA levels of the well-established tonicity-sensitive genes coding for the aldose reductase (AR), the sodium myo-inositol transporter (SMIT) and the betaine transporter (BGT1) were also determined for the sake of comparison. In normally hydrated rats, TauT-, CDO-, and CSD-mRNA were enriched in the outer stripe of the outer medulla (OS). Following antidiuresis, the mRNA levels of TauT, CDO, CSD, SMIT, BGT1 and AR were all similarly increased in the papilla when compared with levels in rats submitted to a chronic diuresis. After acute salt loading, the mRNA level of TauT, like that of SMIT and BGT1, was overexpressed in OS whereas the mRNA levels of CDO and CSD remained unchanged. Like SMIT, BGT1 and AR genes, TauT, CDO and CSD genes appear to be tonicity-sensitive genes which can be activated in vivo by hypertonicity in the rat kidney. However, tonicity-induced activation of the TauT gene is more sensitive than that of CDO and CSD genes.

Aldehyde Reductase↗

PubMed2001

Influence of dehydration on glycerophosphorylcholine and choline distribution along the rat nephron.

Glycerophosphorylcholine is one of the four major organic osmolytes in renal medullary cells, changing their intracellular osmolyte concentration in parallel with extracellular tonicity during cellular osmoadaptation. In this study, the tubular content of glycerophosphorylcholine was quantified in untreated and 48-h-dehydrated male rats. A chemiluminescence ultra-micromethod was developed to measure choline at the picomolar level in single tubules microdissected from collagenase-treated kidneys. The glycerophosphorylcholine level was calculated as the difference between total choline after acid hydrolysis and the free tubular choline content. In accordance with the glycerophosphorylcholine distribution pattern in different renal zones of untreated rats, low amounts of glycerophosphorylcholine were found in all cortical and outer medullary structures (< 35 pmol/mm), whereas increasing amounts were detected towards the papillary tip (163 pmol/mm). As a percentage of total choline, the level of free tubular choline varied from 4.2% in outer medullary proximal tubules to 30.3% in the inner medullary collecting ducts adjacent to the outer medulla (IMCD1). Antidiuresis led to a nearly twofold increase in glycerophosphorylcholine content in papillary collecting ducts. The osmolality-dependent regulation of organic osmolytes in single microdissected tubules has been demonstrated for the first time. Furthermore, the high tubular glycerophosphorylcholine concentration compared to sorbitol and myo-inositol emphasizes the predominance of glycerophosphorylcholine in the inner medulla and papilla of the rat kidney.

Animals↗

PubMed2001

Influence of 72% injury in one kidney on several organs involved in guanidino compound metabolism: a time course study.

Arginine (Arg) produced from citrulline originates mostly from kidneys. Arg is involved in guanidino compound biosynthesis, which requires interorgan co-operation. In renal insufficiency, citrulline accumulates in the plasma in proportion to renal damage. Thus, disturbances in Arg and guanidino compound metabolism are expected in several tissues. An original use of the model of nephrectomy based on ligating branches of the renal artery allowed us to investigate Arg and guanidino compound metabolism simultaneously in injured (left) and healthy (right) kidneys. The left kidney of adult rats was subjected to 72% nephrectomy. Non-operated, sham-operated and nephrectomized rats were studied for a period of 21 days. Constant renal growth was observed only in the healthy kidneys. Guanidino compound levels were modified transiently during the first 48 h. The metabolism and/or tissue content of several guanidino compounds were disturbed throughout the experimental period. Arg synthesis was greatly reduced in the injured kidney, while it increased in the healthy kidney. The renal production of guanidinoacetic acid decreased in the injured kidney and its urinary excretion was reduced. The experimentally proven toxins alpha-keto-delta-guanidinovaleric acid and guanidinosuccinic acid (GSA) accumulated only in the injured kidney. The urinary excretion of GSA and methylguanidine increased in nephrectomized rats. When the injured kidney grew again, the level of some guanidino compounds tended to normalize. Nephrectomy affected the guanidino compound levels and metabolism in muscles and liver. In conclusion, the specific accumulation of toxic guanidino compounds in the injured kidney reflects disturbances in renal metabolism and function. The healthy kidney compensates for the injured kidney's loss of metabolic functions (e.g. Arg: production). This model is excellent for investigating renal metabolism when a disease destroys a limited area in one kidney, as is observed in patients.

Acute Kidney Injury↗

PubMed2000

Ornithine metabolism along the female mouse nephron: localization of ornithine decarboxylase and ornithine aminotransferase.

The fate of ornithine in the nephron of the female OF-1 Swiss mouse remains unknown. The aim of the present study was to identify the nephron segments containing the key enzymes involved in ornithine metabolism: ornithine decarboxylase (ODC) and ornithine aminotransferase (OAT). Viable tubules isolated by microdissection were incubated with [1-14C]ornithine to study the oxidative pathway. Other tubules were permeabilized to measure the ODC activity. Ornithine was decarboxylated in all intact tubules. Gabaculine, a suicide inhibitor of OAT, and rotenone sharply decreased the production of 14CO2 from [1-14C]ornithine. No ODC activity was found in permeabilized tubules isolated from untreated mice. Testosterone increased ODC activity in the proximal tubule substantially and to a minor extent in other nephron segments. In situ hybridization showed ODC messenger ribonucleic acid (mRNA) to be absent in kidneys of untreated females but abundant in the cortex and the outer stripe of the outer medulla of testosterone-treated female mice. The whole proximal tubule contained a great density of silver grains corresponding to ODC mRNA. In conclusion, no basal ODC activity was found in the nephron of female mice. The testosterone-inducible ODC is localized mainly in the proximal tubule, but is also present in distal tubules and collecting ducts. OAT is distributed along the whole nephron, but its activity is higher in proximal tubules than in distal tubules.

Animals↗

PubMed2000

NG, NG-dimethylarginine and NG, NG-dimethylarginine in renal insufficiency.

Asymmetric NG,NG-dimethylarginine (ADMA) and symmetric NG,NG-dimethylarginine (SDMA) are basic endogenous amino acids with a guanidino group. Our renal distribution study of dimethylarginines clearly indicates that, in mouse and rat, ADMA and SDMA levels are most abundant as protein-incorporated compounds (95%). ADMA represents almost 90% of this protein-incorporated dimethylarginine amount. The four zones studied (cortex, outer and inner stripe of outer medulla, inner medulla) contain more or less the same amount of protein-incorporated dimethylarginine; the concentrations of both free dimethylarginines vary more in the different zones. Plasma and urinary excretion levels in Man, rat and mouse were determined, their changes in renal insufficiency were examined and compared between species. Highly significant negative correlations between both plasma dimethylarginine levels and creatinine clearances were found in Man and rat. The correlation between urinary ADMA excretion levels and creatinine clearances was highly significant and positive in Man and mouse; however, in rat the correlation was negative. In patients with severe renal insufficiency, ADMA clearance was only 9.5% of controls, and that of SDMA only 7.8%. Clearance of ADMA and SDMA in nephrectomized mice was 60.5% and 53.8% of controls, respectively, whereas in nephrectomized rat, ADMA clearance actually increased 5.4 times and that of SDMA did not change significantly. Man, rat and mouse show similarities as well as differences in metabolism.

Aged↗

PubMed2000

Polyamines are unevenly distributed within the rat and rabbit kidney.

Aliphatic polyamines have generally been measured on the whole kidney. Since the kidney is composed of a variety of cells, whole organ data are of limited value for the interpretation of the functions of the polyamines. The aim of this study was to establish the distribution pattern of putrescine, spermidine and spermine within the kidneys of male and female rats and rabbits. It is shown that the polyamines are unevenly distributed along the cortico-papillary axis. Each amine exhibited its own distinct distribution pattern. The polyamines are predominantly located in the cortex. Putrescine levels increased gradually from the cortex to the papillary tip in rabbits, whereas, in rats, fluctuations in putrescine level were marked. In the six zones of the rabbit kidney studied, spermidine and spermine concentrations were markedly higher in females than in males. This difference was less marked in rats.

Animals↗

PubMed2000

Regional expression and histological localization of cysteine sulfinate decarboxylase mRNA in the rat kidney.

Cysteine sulfinate decarboxylase (CSD) is the rate-limiting biosynthetic enzyme of the pathway that forms taurine, a putative osmolyte in the kidney, which was previously localized in various segments of the nephron. Although CSD is known to be expressed in whole kidney extracts, no information on CSD mRNA regional expression and histological localization is yet available. Western blotting and Northern blotting were performed in four dissected regions of the kidney using an antiserum against recombinant CSD and a [(32)P]-dCTP-labeled CSD cDNA probe, respectively. In situ hybridization was carried out using a [(35)S]-CTP-labeled CSD RNA probe. A single protein (53 kD) and a single mRNA (2.5 kb) were detected, both of which appeared to be most enriched in the outer stripe of the outer medulla. In situ hybridization of CSD mRNA showed strong labeling of the thick tubules in the outer stripe of the outer medulla and in cortical medullary rays that corresponded to the proximal straight tubules. The significance of this restricted expression of CSD is discussed in relationship to the data previously reported on the location of taurine and the location of the taurine transporter along the nephron.

Animals↗

PubMed1999

Identification of two arginase isoenzyme activities along the nephron of Meriones shawi.

Conflicting theories on the existence of several renal arginase isoenzymes remain in debate. Because the activity of arginase is high in two embryologically different nephron segments of the Meriones shawi kidney, namely the cortical (CPST) and medullary (OSPST) proximal straight tubule and the outer medullary collecting duct (OMCD), we postulate that these nephron segments may contain different isoforms. Isolated nephron segments were dissected from collagenase-treated kidneys. Tubules were permeabilized with Triton X-100 (0.25%) and incubated with increasing Arg concentrations to characterize the arginase activity. The results were as follows: (1) in OMCD, one arginase isoform (E1), characterized by a high Arg affinity (1.160 mM), was present; (2) in CPST, two arginase isoforms were discovered - one, E1, had a similar Km (1.407 mM) to that found in OMCD whereas the other (E2) had a low affinity for Arg (Km =18.8 mM); and (3) in OSPST, two isoenzymes were present - E1 which had a Km of 1.478 mM and the second isoform that we named E2 which had a Km of 9.07 mM. In addition, arginase located in CPST and OMCD was strongly inhibited by Orn and Lys. The Ki value for Lys varied between 1.635 and 2.288 mM. Therefore, this work demonstrates that two arginase isoforms are present in the kidney of Meriones shawi. Isoform E1 is present in the proximal tubule and the collecting duct whereas isoform E2 is restricted to the proximal tubule.

Animals↗

PubMed1998

Biochemical and histopathological changes in nephrectomized mice.

Renal failure is characterized by the retention of nitrogenous metabolites such as urea, creatinine (CTN) and other guanidino compounds (GCs), uric acid, and hippuric acid, which could be related to the clinical syndrome associated with renal insufficiency. A model of renal failure has been developed in male C57BL x Swiss-Webster mice using nephrectomy (NX) and/or arterial ligation. A sham group (group A) and two nephrectomized groups, group B (one kidney removed) and group C (one kidney removed and ligation of the contralateral anterior artery branch), were studied. Ten days postsurgery, morphological and functional indices of renal failure were investigated. Nephrectomized mice manifested features of renal failure like polyuria and wasting. CTN clearance (CTN[Cl]) decreased by +/-26% in group B and +/-33% in group C as compared with the control values. Marked increases in the plasma concentration of guanidinosuccinic acid ([GSA] fourfold) and guanidine ([G] twofold) were observed in the experimental animals. CTN and alpha-keto-delta-guanidinovaleric acid (alpha-keto-delta-GVA) reached levels of, respectively, 1.5-fold and twofold those of controls. Urinary GSA excretion increased and guanidinoacetic acid (GAA) excretion decreased about twofold in group C. GSA increases (2.6-fold) were also observed in the brain in group C, in addition to a significant increase of G (2.5-fold) and gamma-guanidinobutyric acid ([GBA] 1.5-fold). Finally, the extent of NX was found to be 45.2% in group B and 71.4% in group C. Light microscopy revealed an expansion and increase in cellularity of the mesangium of the glomeruli, particularly in group C. A significant correlation (r = .574, P < .0001) was found between CTN(Cl) and the degree of NX as calculated from the remaining functional area. These data suggest that the model can be used as a tool for further pathophysiological and/or behavioral investigations of renal failure.

Animals↗

PubMed1998

Ornithine decarboxylase along the mouse and rat nephron.

Renal arginase activity is a potent source of ornithine (Orn) for polyamine synthesis. Ornithine decarboxylase (ODC) was localized along the mouse and rat nephron by incubating viable nephron segments isolated by microdissection from collagenase-treated kidneys with or without D,L-2-(difluoromethyl)ornithine (DFMO), a selective inactivator of ODC. Tubules from either control or DFMO-treated animals were incubated with 100 ¿M L-[1-14C]Orn. In control mice, Orn decarboxylation occurred mainly in the proximal convoluted tubule (PCT). In DFMO-treated mice, Orn decarboxylation was dramatically reduced in PCT and in proximal straight tubules (PST). In rats, Orn decarboxylation also occurred predominantly in the proximal tubule. Addition of 10 mM DFMO to isolated tubules dramatically decreased Orn decarboxylation in PCT and in PST. Thereafter, ODC activity was demonstrated in permeabilized tubules. In Triton X-100-treated tubules from control mice, ODC was exclusively found in proximal tubules (PCT > PST). This ODC activity was strongly inhibited in DFMO-treated mice. In conclusion, the highest ODC activity was found in rat and mouse PCT, a segment devoid of arginase. We hypothesize that the filtered Orn, which is reabsorbed along the PCT,is the main source of Orn for ODC.

Animals↗

PubMed1997

Renal handling of guanidino compounds in rat and rabbit.

1. Guanidino compounds (GCs) have been quantified in different mammalian tissues such as brain, liver, muscle and kidney. The high anatomical heterogeneity of the kidney suggests that GCs could be unevenly distributed along the corticopapillary axis of the kidney in different species. 2. This study was designed to quantify twelve GCs in the different zones of rat and rabbit kidney. The kidneys were sliced and pieces of seven definite zones were weighed and homogenized for further GC extraction. GCs were determined by liquid chromatography. 3. The results indicate that: (1) GCs were unevenly distributed along rat and rabbit kidney; (2) qualitative and quantitative studies proved that each GC shows a particular distribution pattern along the corticopapillary axis for a given species; (3) in rats, alpha-keto-delta-guanidinovaleric acid, guanidinosuccinic acid, creatinine (CTN), methylguanidine and to a lesser extent gamma-guanidinobutyric acid increased steeply along the inner medulla in parallel to urea, whereas in rabbits, most of the GCs reached a plateau in the inner medulla and remained constant at this level; (4) gamma-guanidinobutyric acid was specifically found in the rat kidney; (5) argininic acid was higher in rabbit compared with rat kidney; (6) significantly higher levels of homoarginine were found in all zones of the rat kidney compared with the rabbit kidney. 4. The results suggest that: (1) GCs are mostly localized within the nephron segments; (2) an accumulation of GCs in the inner medulla might be explained either by a recycling process or by an intracellular storage as has been reported for urea, amino acids and organic osmolytes; (3) some GCs might be synthesized in nephron segments as reported for arginine (Arg) and guanidinoacetic acid (GAA); (4) several metabolic pathways of the GCs seemed to differ between rat and rabbit; (5) except for creatine, CTN, Arg and GAA, it seems unlikely that GCs might significantly increase the intracellular osmolality.

Affinity Labels↗

PubMed1997

Amino acid handling in uremic rats: citrulline, a reliable marker of renal insufficiency and proximal tubular dysfunction.

The kidney is involved in amino acid reabsorption and metabolism; consequently, in renal insufficiency, these important functions are disturbed, as has been reported in animals and patients. In a first experimental series, rats were subjected to degrees of nephrectomy (NX) varying between 10% and 90%. Three weeks later, amino acid levels were measured in plasma to correlate the levels with the degree of NX. The results indicate that in the range of 33% to 74% NX, the plasma concentration of only three to four amino acids was modified, whereas in rats with 84% NX, the concentration of 11 amino acids was disturbed, compared with sham-operated rats. Citrullinemia was enhanced in uremic rats and correlated with the degree of NX. More interestingly, citrullinemia was increased in the range of 10% to 33% NX without any changes in uremia and creatininemia, two well-known markers of uremic states. A second experimental series was designed to study the time course of changes in aminoacidemia to find a marker for the onset of renal failure. Rats were subjected to 36% NX for a period of 1 to 21 days. Uremia and creatininemia peaked 24 to 48 hours after NX, and creatinine clearance (Clcreat) concomitantly diminished. Unfortunately, these three markers of uremic states returned to control values during the next few days before increasing during the last 2 weeks. In contrast, citrullinemia increased twofold 48 hours after NX and plateaued over the next 20 days. We conclude that in rats, citrullinemia could be used (1) to detect acute and chronic renal failure, (2) as a specific marker of normal function of the proximal tubule, and (3) to estimate the degree of renal damage. From this study, renal insufficiency might be easily detected by measuring citrullinemia.

Amino Acids↗

PubMed1996

Arginine metabolism in cat kidney.

1. Arginine is essential for growth in the kitten and, because of the resulting hyperammonaemia, in the adult cat an arginine-free diet is life threatening. 2. The kidney is the main site of arginine synthesis. 3. This study was performed to determine whether the cat kidney synthesizes arginine and to establish which factors, such as low citrullinaemia, defects of argininosuccinate synthase and lyase activities or high renal arginase activity, might limit renal arginine production. 4. Identified nephron segments were isolated by microdissection from collagenase-treated cat kidney. 5. Arginine metabolism was studied by incubating the nephron segments with either physiological concentrations of L-[ureido-14C]citrulline (anabolism) or L-[guanido-14C]-arginine (catabolism). Arginine and urea were measured by a micro-enzymatic method. Amino acids were measured by HPLC. 6. In cat blood, the citrulline, but not the arginine, concentration was very low by comparison with other species. 7. Arginine synthesis occurred almost entirely in the proximal tubule, the highest rate occurring in the proximal convoluted tubule and the lowest in the medullary straight proximal tubule. 8. Arginase activity was restricted to the proximal tubule. Urea production increased from the convoluted towards the medullary straight tubule. 9. The limited capacity of the cat kidney to produce arginine in vivo may result from the low blood concentration of citrulline and from the high arginase activity in the various proximal cells with the ability to synthesize arginine.

Animals↗

PubMed1996

Biochemical heterogeneity of arginine metabolism along kidney proximal tubules.

By using an in vitro single tubule micromethod of high specificity, in four different species of mammals it has been observed that (a) arginine synthesis from citrulline (arginine synthase activity, E.C. 6.3.4.5 and E.C. 4.3.2.1) is restricted to the early portions of proximal convoluted tubules, whereas (b) urea production from arginine (arginase activity, E.C. 3.5.3.1.) is present mainly in the cortical (CPST) and even more in the outer medullary (OSPST) portions of straight proximal tubules. The data suggest that (a) in early PCT cells, the citrulline reabsorbed from glomerular filtrate is converted into arginine, which in turn crosses peritubular cell membranes together with reabsorbed arginine, and (b) the urea formed in CPST and OSPST cells might passively diffuse into the luminal fluid entering Henle's loops. Such urea secretion might contribute to sustain the process of urea recycling in kidney medulla and thereby participate in the mechanism of urine concentration.

Animals↗

PubMed1995

Sites of arginine synthesis and urea production along the nephron of a desert rodent species, Meriones shawi.

The distribution of arginine synthase and arginase activities along the successive nephron segments of Meriones kidney was measured in vitro with single tubule enzymatic microtechniques making use of either L-[ureido-14C] Citrulline (0.108 mM) or L-[guanidino-14C]arginine (0.2 mM) as the respective substrates. Arginase activity (fmol urea formed per min per mm of tubule) was very low (5-25 fmol.min-1.mm-1) in most nephron segments including the early portions of proximal convoluted tubules (early PCT). It increased progressively after 3 mm of the PCT to reach a value of 200 fmol.min-1.mm-1 in the cortical portion of the straight proximal tubule (CPST), with a further increase, along the pars recta, of up to 250 fmol.min-1.mm-1 in the outer medullary portion (OSPST). In addition, arginase activity in OSPST and the adjacent descending thin limb (DTL) was higher in juxtamedullary nephrons (JN) than in the corresponding portions of superficial nephrons (SN). Arginine synthase activity (fmol arginine formed per mm of tubule per min) was present in proximal tubules exclusively, with a gradient decreasing along the PCT (about 600 fmol.min-1.mm-1 in the 1st mm, 65 fmol.min-1.mm-1 in CPST and 30 fmol.min-1.mm-1 in OSPST). It has been checked that CPST and OSPST (where the two enzyme systems are present) are able to convert citrulline directly into urea with a yield of 65%.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1995

Guanidino compound metabolism in rats subjected to 20% to 90% nephrectomy.

In mammalian kidney, the proximal convoluted tubule (PCT) is the main site of arginine (Arg) production. Arginine can be used in the biosynthesis of guanidino compounds (GC). Since uremic rats have a lower functional mass of PCT, GC synthesis might be modified, especially that of guanidinoacetic acid (GAA) which occurs in PCT. In order to study GC metabolism at different steps of uremia, rats were subjected to either 42% or 80% nephrectomy (NX); the experiment lasted for three weeks. Results show that: (1) in plasma, the pattern of GC levels in 42% NX rats was similar to that of controls except for a clear increase of beta-guanidinopropionic acid (beta-GPA), whereas in 80% NX rats, all GC levels sharply increased except that of creatine which decreased. (2) Urinary excretion of GC in control and 42% NX rats is quite similar except for GAA which strongly decreased, and for homoarginine (HArg) and argininic acid (ArgA) which increased. In rats with 80% NX, the principal modification in GC excretion was a four- to five-fold reduction in GAA output. (3) After induction of renal failure, Arg, creatine and guanidinosuccinic acid reabsorption remained unchanged, and that of HArg decreased. For guanidine and methylguanidine the negative renal balance remained unchanged, and that of gamma-guanidinobutyric acid, GAA and alpha-keto-delta-guanidinovaleric acid became smaller, suggesting a better reabsorption. In conclusion, uremia strongly modified GC metabolism involving mainly those synthesized from Arg; both GAA and creatine synthesis were strongly decreased probably because of the loss of renal tissue, mainly PCT.

Animals↗

PubMed1994

Urea production by kidney collecting ducts in vitro: effect of amino acid addition.

Urea production by cortical (CCD) and medullary (OMCD) collecting ducts of the rat kidney was measured in vitro by incubating single microdissected pieces of tubule in the presence of L-[guanido-14C]arginine (0.2 mM). The [14C]urea released from the cells was hydrolysed in presence of urease added to the incubation medium and the 14CO2 formed was trapped in KOH and counted. The effect of various amino acids (AA) on urea production was investigated by adding unlabelled AA (either in combination or singly) at concentrations close to those present in blood plasma. A mixture of 17 AA decreased urea production from [14C]arginine by 46% in CCD and by 58% in OMCD. When lysine and proline were omitted from the mixture, the inhibition was less marked (19% in CCD and 43% in OMCD, respectively). When AA were tested singly, lysine induced the larger inhibition (40% in CCD and 45% in OMCD), than ornithine and glutamine (about 15% each, in CCD and OMCD), whereas proline inhibition (7% in CCD, 10% in OMCD) was not statistically significant. Branched-chain amino acids (BCAA) in combination (leucine, isoleucine and valine) also markedly reduced urea production by CCD and OMCD. Their effect was dose dependent. Solubilization of CCD and OMCD cell membranes with Triton X-100 resulted in a twofold increase in urea production by control samples; the relative inhibition (per cent) induced by BCAA was enhanced, whereas that induced by lysine was decreased. The data suggest that, in living tubules, the inhibition obtained with lysine resulted, for a large part, from competition between lysine and arginine for cell uptake via a common membrane carrier, whereas the inhibition induced by BCAA corresponded to an effect on arginase activity itself.

Amino Acids↗

PubMed1993

Valine oxidation in the rat medullary thick ascending limb.

In the kidney, a branched-chain amino acid transferase (BCAAT) activity has been localized mainly in the medullary thick ascending limb (MTAL) of the rat nephron. BCAAT is the first enzyme involved in the metabolic pathway of the three branched-chain amino acids (BCAA): leucine, isoleucine and valine. The present work has been designed to study valine catabolism. Valine and leucine oxidation in MTAL were compared by measuring the rate of 14CO2 produced when these substrates were incubated as sole substrates at a final concentration of 1 mM. Since glucose is also metabolized in MTAL, valine and leucine oxidation were quantified also in the presence of glucose (1 mM). The results show that: (1) valine oxidation was greater than that of leucine (63.0 +/- 4.7 vs 39.7 +/- 5.2 pmol.h-1 x micrograms-1 protein, respectively; P < 0.001). As previously shown, leucine oxidation was found to be increased in the presence of glucose whereas glucose oxidation decreased. In contrast, the presence of glucose strongly diminished valine oxidation (19.2 +/- 1.9 vs 63.1 +/- 4.7 pmol.h-1 x micrograms-1 protein; P < 0.001) whereas glucose oxidation was increased in the presence of valine (268.2 +/- 14.9 vs 229.6 +/- 16.2 pmol.h-1 x micrograms-1 protein; P < 0.05). We conclude that in rat MTAL, under near physiological conditions (in the presence of glucose, as in vivo), leucine is a preferential respiratory fuel as compared to valine. However, valine supports energetic salt transport and facilitates glucose oxidation.

Animals↗