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G T Nagami

Publications and source records attributed to G T Nagami.

25 records · Page 2Linked to original sources
PubMed1986

Ammonia production by isolated mouse proximal tubules perfused in vitro. Effect of metabolic acidosis.

We examined the effects of metabolic acidosis in vivo and reduced bath and luminal pH in vitro on total NH3 (NH3 + NH+4) production rates by isolated mouse proximal tubule segments. Midproximal tubule segments were obtained from mice with NH4Cl-induced metabolic acidosis and from nonacidotic controls. The segments were perfused with modified Krebs-Ringer bicarbonate (KRB) buffer, incubated in KRB buffer containing 0.5 mM L-glutamine and 1.0 mM sodium acetate, and gassed with 95% O2 and 5% CO2. Isolated unperfused and perfused proximal tubules from acidotic mice produced total NH3 at higher rates than corresponding tubules from nonacidotic mice. Perfusion of the tubular lumen stimulated total NH3 production by tubules from both acidotic and nonacidotic mice. In contrast, lowering the bath pH to 7.0 by lowering the HCO3- concentration increased total NH3 production rates by tubules from nonacidotic mice but not by tubules from acidotic mice. Reducing the HCO3- concentration of the bath buffer to 10 mM while maintaining a pH of 7.4 had no significant effect on total NH3 production by tubules from nonacidotic mice. Lowering the luminal fluid pH by reducing the perfusate HCO-3 from 25 mM to 10, 5, or 1.2 mM while maintaining a bath pH of 7.4 lowered collected luminal fluid pH but had no effect on total NH3 production by proximal tubules from nonacidotic mice. These observations demonstrated that metabolic acidosis in vivo stimulated total NH3 production in isolated mouse proximal tubule segments and that low peritubular pH and HCO-3 stimulated total NH3 production by proximal tubule segments from nonacidotic mice in vitro.

Acidosis↗
PubMed1985

Regulation of ammonia production by mouse proximal tubules perfused in vitro. Effect of luminal perfusion.

To investigate factors regulating ammonia (NH3) production by isolated defined proximal tubule segments, we examined the rates of total NH3 (NH3 + NH+4) production by individual proximal tubule segments perfused in vitro under a variety of perfusion conditions. Segments consisting of late convoluted and early straight portions of superficial proximal tubules were incubated at 37 degrees C in Krebs-Ringer bicarbonate (KRB) buffer containing 0.5 mM L-glutamine and 1.0 mM sodium acetate, pH 7.4. The rate of total ammonia production was calculated from the rate of accumulation of total NH3 in the bath. The total ammonia production rate by unperfused proximal segments was 6.0 +/- 0.2 (+/- SE) pmol/mm per minute, which was significantly lower than segments perfused at a flow rate of 22.7 +/- 3.4 nl/min with KRB buffer (21.5 +/- 1.4 pmol/mm per minute; P less than 0.001) or with KRB buffer containing 0.5 mM L-glutamine (31.9 +/- 2.5; P less than 0.001). The rate of NH3 production was higher in segments perfused with glutamine than in segments perfused without glutamine (P less than 0.01). The perfusion-associated stimulation of NH3 production was characterized further. Analysis of collected luminal fluid samples revealed that the luminal fluid total NH3 leaving the distal end of the perfused proximal segment accounted for 91% of the increment in NH3 production observed with perfusion. Increasing the perfusion flow rate from 3.7 +/- 0.1 to 22.7 +/- 3.4 nl/min by raising the perfusion pressure resulted in an increased rate of total NH3 production in the presence or absence of perfusate glutamine (mean rise in rate of total NH3 production was 14.9 +/- 3.7 pmol/mm per minute in segments perfused with glutamine and 7.8 +/- 0.9 in those perfused without glutamine). In addition, increasing the perfusion flow rate at a constant perfusion pressure increased the rate of luminal output of NH3. Total NH3 production was not affected by reducing perfusate sodium concentration to 25 mM and adding 1.0 mM amiloride to the perfusate, a condition that was shown to inhibit proximal tubule fluid reabsorption. These observations demonstrate that the rate of total NH3 production by the mouse proximal tubule is accelerated by perfusion of the lumen of the segment, by the presence of glutamine in the perfusate, and by increased perfusion flow rates. The increased rate of NH3 production with perfusion seems not to depend upon normal rates of sodium reabsorption. The mechanism underlying the stimulation of NH3 production by luminal flow is unknown and requires further study.

Absorption↗
PubMed1985

Cytosolic redox potential and phosphate transport in the proximal tubule of the rabbit. A study in the isolated perfused tubules.

Recently it has been proposed that cytosolic NAD+ may play a regulatory role in phosphate transport across the renal proximal tubules. To test this thesis, we have examined the effect of altering the cytosolic redox potential (NAD+/NADH) on phosphate flux across the isolated perfused rabbit proximal tubules. The cytosolic redox potential was shifted to a more oxidized state either by changing the substrate in the bathing medium from lactate to pyruvate or by adding methylene blue to the bath medium. On the other hand, cytosolic redox potential was shifted to a more reduced state by changing the substrate in the bath medium from pyruvate to lactate. In either case the phosphate flux across the proximal tubule was unaffected. It was concluded that cytosolic redox potential may not play a significant role in regulating the phosphate transport in renal proximal tubules.

Animals↗
PubMed1984

Dissociation of gluconeogenesis from fluid and phosphate reabsorption in isolated rabbit proximal tubules.

Gluconeogenesis in the kidney is a metabolic function characteristic of proximal tubules. Recent studies suggest that renal gluconeogenesis (GNG) may in some way be coupled to fluid and phosphate reabsorption in the proximal tubule. Therefore, the present studies examined more directly the relationship between GNG and transport of fluid and phosphate using isolated proximal straight tubules of rabbit kidney. Glucose production rates were determined in isolated tubules with a glucose microassay while both phosphate (Jp) and net fluid fluxes (Jv) were measured by the in vitro isolated tubule perfusion technique. Glucose production rates from individual substrates, including pyruvate, lactate, glutamate and alpha-ketoglutarate, differed significantly, but neither Jv nor Jp was altered when different substrates in the bath medium were used. Inhibition of GNG by 3-mercaptopicolinate did not alter the Jv or Jp. Acid pH (7.0) stimulated GNG and suppressed both Jv and Jp. The addition of 3-mercaptopicolinate at acid pH abolished the stimulatory effect of acid pH on GNG but had no effect on Jv or Jp. These data thus indicate that, in the isolated rabbit renal proximal tubule, GNG rates can be dissociated from both the Jv and Jp and suggest that renal GNG may not directly regulate the fluid and phosphate transport rates in the proximal straight tubule.

Animals↗