Effect of D-glucose and L-glutamate on the kinetics of bicarbonate reabsorption in rat proximal tubules

The effect of D-glucose or L-glutamate on the kinetics of bicarbonate reabsorption in the early (EPT) and middle proximal tubule (MPT) was studied in vivo in Munich-Wistar rats by microperfusion techniques. The presence of 20 mM D-glucose in the lumen increased acidification half-time (t/2) (from 2.54 + or - 0.09s to 3.11 + or - 0.17 s in EPT and from 4.75 + or - 0.20 s to 6.04 + or - 0.49 s in MPT). Bicarbonate reabsorption 9JHCO-3) decreased as a consequence of this change (from 3.80 + or - 0.17 to 2.46 + or - 0.20 nmol cm-2s-1 in EPT and from 2.30 + or - 0.10 to 1.64 + or - 0.10 nmol cm-2s-1 in MPT). In this situation the basolateral membrane potential difference (BLMPD) in the MPT decereased from -41.6 + or - 2.47 to -29.7 + or - 2.45 mV and returned to control values after perfusion with D-glucose. The addition of 20 mM L-glutamate to the luminal perfusion caused an opposite effect, i.e., a decrease in t/2 (1.54 + or - 0.21 s in EPT and 3.25 + or - 0.26 s in MPT) and a consequent increase in JHCO-3 in both segments (5.09 + or - 0.58 nmol cm-2s-1 in EPT and 3.92 + or - 0.30 nmol cm-2s-1 in MPT). The BLMPD of MPT increased during L-glutamate perfusion (-39 + or - 2.48 mV in control and -52.0 + or - 2.72 mV with L-glutamate) and returned to control values after perfusion. The results observed may be the consequence of coupled Na+ - substrate transport altering the BLMPD with modifies the electrical driving force for coupled Na+ -HCO-3 efflux across the basolateral membrane. The alteration of this process may in turn affect intracellular pH, which is an important modulator of luminal Na+./H+ enchange. This possibility is supported by the observed depolarization of BLMPD by D-glucose (electroneutral molecule), and hyperpolarization by glutamate anion

Atrial natriuretic peptide modulates the effect of angiotensin II on the concentration of free calcium in the cytosol of Mandin-Darby canine kidney cells

The effect of angiotensin II (ANG II) and atrial natriuretic peptide (ANP) on intracellular free calcium concentration [ Ca²+]i was investigated in Mandin-Darby canine kidney (MDCK) cells in culture. Changes in [Ca²+]i were monitored fluorometrically with the Ca²+ -sensitive probel fura -2/AM at 37ºC using Perkin-Elmer LS-5 spectrofluorimeter (excitation 340/380 nm,slite 3 nm; emission 520 nm, slit 10 nm). MDCK cells exhibited a mean baseline [Ca²+]i of 98 ñ 10 nM. the addition of increasing concentrations of SNG II (1 pM to 1 µM) to the cell suspension led to a progressive increase in [Ca²+]i to 2-3 times basal levels. In contrast, addition of 1 µM ANP to the cell suspension led to a very rapid 60 percent decrease in [Ca²+]i. The addition of 1 pM to 1 µM ANG II immediately after 1 µM ANP caused an increase in [Ca²+]i which never exceded the basal level in the absence of ANP. The data indicate that ANG II increases cell [Ca²+]i as expected, and provide the new observation that ANP reduces [Ca²+]i in these cells. Further more, ANP reduces the increase in [Ca²+]i elicited by ANG II, thus modulating the effect of ANG II on [Ca²+]i

Effect of glucose on the kinetics of bicarbonate reabsorption in the early and middle proximal tubule

The kinetics of bicarbonate reabsorption in the early (EPT) and middle proximal tubule (MPT) was studied in Munich Wistar rats by means of microperfusion techniques. The early proximal tubule (EPT) had a higher capacity of reabsorbing bicarbonate than the middle segment. The calculated bicarbonate flow (JHCO3) was 4.47 nmol s**-1 cm**-2 int the EPT and 2.30 nmol s**-1 cm**-2 in the MPT and the half time of injected bicarbonate (t/2) was less in the EPT (2.24 s) than in the MPT (5.20 s). The presence of 20 mM glucose in the lumen led to a reduction in the velocity of tubular acidification in the EPT (3.11 s) and caused a decrease in JHCO3 in both segments (2.46 nmol s**-1 cm**-2 in the EPT and 1.64 nmols s**-1 cm**-2 in the MPT). These alterations may be the result either of lumen-to-cell sodium gradient dissipation or electrical changes induced by sodium-glucose cotransport that may depolarize the basolateral membrane leading to intracellular alkalinization. This effect may then impair the Na +/H+ exchanger, thus decreasing bicarbonate reabsorption

Impairment of the renal tubular acidification kinetics by lithium

We studied the kinetics of HCO-3 reabsorption in the middle proximal (MPT) and distal convoluted tubules (DCT) by measuring continuously intratubular pH with Sb-microelectrodes in stopped-flow microperfusion (HCO-3, 30 mM Ringer) experiments. Male Wistar rats (240-280 g) were injected ip with LiCl (4mEqkg-1 day-1) for 4 days (Li) and were compared to controls (C). Steady-state pH increased in MPT from 6.64 + or - 0.02(57) to 6.89 + or - 0.02(45), mean + or - SEM (number of measurements) on tissue from 13 rats in each group, and from 6.87 + or - 0.5(30) to 7.08 + or - 0.01(63) in DCT. HCO-3 reabsorption decreased from 1.32 + or - 0.08(57) to 0.96 + or - 0.4(45) nmol cm-2 s-1 in MPT and from 0.85 + or - 0.07(30) to 0.45 + or - 0.04(63) in DCT. These data indicate that lithium affected the acidification mechanism in MPT and DCT, probably through an impairment of the Na+ -H+ antiport in both tubular segments