Axial heterogeneity in the rat proximal convoluted tubule. II. Osmolality and osmotic water permeability

1984 ◽  
Vol 247 (5) ◽  
pp. F822-F826 ◽  
Author(s):  
F. Y. Liu ◽  
M. G. Cogan ◽  
F. C. Rector

To assess whether proximal luminal fluid becomes hypotonic with respect to plasma, free-flow micropuncture measurements were made sequentially from the end-proximal tubule to Bowman's space in 10 tubules of hydropenic Munich-Wistar rats. Osmolality in Bowman's space was 2.8 +/- 0.3 mosmol less than in plasma. Tubular fluid osmolality fell along the tubule and by the end-proximal tubule was 7.5 +/- 0.7 mosmol/kg less than in plasma or 4.7 mosmol/kg less than in Bowman's space. Since luminal fluid became hypotonic, the reabsorbate was hypertonic. The transepithelial osmotic water permeability (Pf) was calculated using simultaneously measured water reabsorption rates. The osmotic gradient responsible for water reabsorption was assumed to be either lumen-to-reabsorbate or lumen-to-peritubular plasma, with a reflection coefficient for sodium chloride of 0.7-1.0. The Pf was then estimated to be between 0.2 and 2.0 cm/s in the first millimeter of tubule and to have fallen to 0.1-0.2 cm/s by the end of the tubule. In conclusion, luminal hypotonicity develops in the rat proximal convoluted tubule and must be considered as part of the osmotic driving force for water reabsorption.

1983 ◽  
Vol 245 (3) ◽  
pp. F279-F294 ◽  
Author(s):  
C. A. Berry

The route of water transport in the proximal tubule could be either transjunctional or transcellular. A transjunctional route is supported by data showing high osmotic-to-diffusive water permeability ratios, the possible correlation of junctional leakiness to ions and nonelectrolytes with water permeability, and solvent drag of nonelectrolytes and ions. These data, however, are not convincing. A transcellular route of water transport is supported by data showing that the osmotic water permeability (Pf) for apical and/or basolateral cell membranes is sufficiently high to account for the transepithelial Pf, making a tentative conclusion for a transcellular route of water transport possible. In addition, measurements of Pf have yielded insights into the mechanism of solute-solvent coupling. Pf has been reported to be mostly between 0.1 and 0.3 cm/s. In the rabbit proximal straight and the Necturus proximal convoluted tubule, in which water transport rates are low, this range of Pf will account for volume absorption with only small osmotic gradients (less than 6 mosmol). Higher osmotic gradients are required in the rat and possibly the rabbit proximal convoluted tubule, where water transport rates are higher. Solute-solvent coupling in all species is probably due to both luminal hypotonicity and lateral intercellular space hypertonicity. These two processes are directly linked. Mass balance requires that generation of luminal hypotonicity also generates a hypertonic absorbate and, thus, some degree of lateral intercellular space hypertonicity. It is likely that, in the rabbit at least, effective osmotic pressure gradients due to differences in solute reflection coefficients play little role in solute-solvent coupling.


2019 ◽  
Vol 316 (2) ◽  
pp. F253-F262 ◽  
Author(s):  
Josephine K. Liwang ◽  
Joseph A. Ruiz ◽  
Lauren M. LaRocque ◽  
Fitra Rianto ◽  
Fuying Ma ◽  
...  

Hypertonicity increases water permeability, independently of vasopressin, in the inner medullary collecting duct (IMCD) by increasing aquaporin-2 (AQP2) membrane accumulation. We investigated whether protein kinase C (PKC) and adenosine monophosphate kinase (AMPK) are involved in hypertonicity-regulated water permeability. Increasing perfusate osmolality from 150 to 290 mosmol/kgH2O and bath osmolality from 290 to 430 mosmol/kgH2O significantly stimulated osmotic water permeability. The PKC inhibitors chelerythrine (10 µM) and rottlerin (50 µM) significantly reversed the increase in osmotic water permeability stimulated by hypertonicity in perfused rat terminal IMCDs. Chelerythrine significantly increased phosphorylation of AQP2 at S261 but not at S256. Previous studies show that AMPK is stimulated by osmotic stress. We tested AMPK phosphorylation under hypertonic conditions. Hypertonicity significantly increased AMPK phosphorylation in inner medullary tissues. Blockade of AMPK with Compound C decreased hypertonicity-stimulated water permeability but did not alter phosphorylation of AQP2 at S256 and S261. AICAR, an AMPK stimulator, caused a transient increase in osmotic water permeability and increased phosphorylation of AQP2 at S256. When inner medullary tissue was treated with the PKC activator phorbol dibutyrate (PDBu), the AMPK activator metformin, or both, AQP2 phosphorylation at S261 was decreased with PDBu or metformin alone, but there was no additive effect on phosphorylation with PDBu and metformin together. In conclusion, hypertonicity regulates water reabsorption by activating PKC. Hypertonicity-stimulated water reabsorption by PKC may be related to the decrease in endocytosis of AQP2. AMPK activation promotes water reabsorption, but the mechanism remains to be determined. PKC and AMPK do not appear to act synergistically to regulate water reabsorption.


Cells ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 2533
Author(s):  
Fuying Ma ◽  
Guangping Chen ◽  
Eva L. Rodriguez ◽  
Janet D. Klein ◽  
Jeff M. Sands ◽  
...  

Adrenomedullin (ADM) is a vasodilator that causes natriuresis and diuresis. However, the direct effect of ADM on osmotic water permeability in the rat inner medullary collecting duct (IMCD) has not been tested. We investigated whether ADM and its ADM receptor components (CRLR, RAMP2, and 3) are expressed in rat inner medulla (IM) and whether ADM regulates osmotic water permeability in isolated perfused rat IMCDs. The mRNAs of ADM, CRLR, and RAMP2 and 3 were detected in rat IM. Abundant protein of CRLR and RAMP3 were also seen but RAMP2 protein level was extremely low. Adding ADM (100 nM) to the bath significantly decreased osmotic water permeability. ADM significantly decreased aquaporin-2 (AQP2) phosphorylation at Serine 256 (pS256) and increased it at Serine 261 (pS261). ADM significantly increased cAMP levels in IM. However, inhibition of cAMP by SQ22536 further decreased ADM-attenuated osmotic water permeability. Stimulation of cAMP by roflumilast increased ADM-attenuated osmotic water permeability. Previous studies show that ADM also stimulates phospholipase C (PLC) pathways including protein kinase C (PKC) and cGMP. We tested whether PLC pathways regulate ADM-attenuated osmotic water permeability. Blockade of either PLC by U73122 or PKC by rottlerin significantly augmented the ADM-attenuated osmotic water permeability and promoted pS256-AQP2 but did change pS261-AQP2. Inhibition of cGMP by L-NAME did not change AQP2 phosphorylation. In conclusion, ADM primarily binds to the CRLR-RAMP3 receptor to initiate signaling pathways in the IM. ADM reduced water reabsorption through a PLC-pathway involving PKC. ADM-attenuated water reabsorption may be related to decreased trafficking of AQP2 to the plasma membrane. cAMP is not involved in ADM-attenuated osmotic water permeability.


1984 ◽  
Vol 247 (5) ◽  
pp. F816-F821 ◽  
Author(s):  
F. Y. Liu ◽  
M. G. Cogan

To measure simultaneously the concentration profiles of bicarbonate, chloride and inulin along the length of the superficial proximal convoluted tubule, free-flow micropuncture measurements were made sequentially from the end-proximal tubule to Bowman's space in 10 tubules of hydropenic Munich-Wistar rats. Bicarbonate and volume reabsorption were 354 +/- 21 pmol X mm-1 X min-1 and 5.9 +/- 0.4 nl X mm-1 X min-1 in the first millimeter and fell progressively in the remaining 3.8 mm of tubule, averaging 83 +/- 4 pmol X mm-1 X min-1 and 2.3 +/- 0.5 nl X mm-1 X min-1, respectively. The values in the initial millimeter represents a high transport capacity since they exceed rates that have been observed when comparable or even higher mean luminal substrate concentrations were presented to the late proximal tubule. In contrast, chloride reabsorption was only 206 +/- 55 peq X mm-1 X min-1 in the first millimeter compared with a mean of 306 +/- 22 peq X mm-1 X min-1 in the rest of the tubule. In conclusion, there is substantial axial transport heterogeneity, with bicarbonate and water reabsorption higher but chloride reabsorption lower in the early compared with the late superficial proximal convoluted tubule.


1985 ◽  
Vol 249 (1) ◽  
pp. F124-F131 ◽  
Author(s):  
P. A. Preisig ◽  
C. A. Berry

To determine the predominant pathway for transepithelial osmotic water flow, the transepithelial osmotic water permeability [Pf(TE)] and the apparent dimensions of paracellular pores and slits were determined in rat proximal convoluted tubules microperfused in vivo. To measure Pf(TE), tubules were perfused with a hyposmotic, cyanide-containing solution. Pf(TE), calculated from the observed volume flux in response to the measured log mean osmotic gradient, was 0.12-0.15 cm/s, assuming sigmaNaCl equal to 1.0-0.7, respectively. The dimensions of the paracellular pathways were determined using measured sucrose and mannitol permeabilities (nonelectrolytes confined to the extracellular space). These were 0.43 and 0.87 X 10(-5) cm/s, respectively. By using the ratio of these permeabilities, their respective free solution diffusion coefficients and molecular radii, and the Renkin equation, the radius of the nonelectrolyte-permeable pores and the total pore area/cm2 surface area/channel length were calculated to be 1.4 nm and 3.56 cm-1, respectively. Similar calculations for slits yielded a slit half-width of 0.8 nm and a total slit area/cm2 surface area/channel length of 3.16 cm-1. The osmotic water permeability of these nonelectrolyte-permeable pathways was calculated by Poiseuille's law to be 0.0018 cm/s (pores) or 0.0014 cm/s (slits), at most 2% of Pf(TE). We conclude that the nonelectrolyte-permeable pathway in the tight junctions is not the major route of transepithelial osmotic water flow in the rat proximal tubule.


1993 ◽  
Vol 264 (3) ◽  
pp. F397-F403 ◽  
Author(s):  
R. Mejia ◽  
M. F. Flessner ◽  
M. A. Knepper

Luminal fluid exiting the proximal convoluted tubule of a juxtamedullary nephron is alkalinized as it passes through the long-loop thin descending limb of Henle (LDL). Three potential mechanisms of alkalinization are: 1) concentration of bicarbonate by water abstraction, 2) direct bicarbonate entry, and 3) NH3 entry. We have used a mathematical model of the LDL to investigate these mechanisms. With permeabilities of HCO3-, NH3, and NH4+ measured for subsegments of the chinchilla LDL [M. F. Flessner, R. Mejia, and M. A. Knepper. Am. J. Physiol. 264 (Renal Fluid Electrolyte Physiol. 33):F388-F396, 1993], the osmotic water permeability of each segment [C.-L. Chou and M. A. Knepper. Am. J. Physiol. 263 (Renal Fluid Electrolyte Physiol. 32):F417-F426, 1992], and appropriate parameters from the literature, we have used the model to calculate hypothetical pH, HCO3- concentration, and NH3 concentration of the luminal fluid as it descends the LDL within an assumed interstitium. After eliminating each mechanism in turn by setting the appropriate permeability to zero, we recalculated the axial profiles. Our results suggest that, although all three mechanisms individually contribute to LDL alkalinization, NH3 entry likely plays the dominant role.


Cells ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 967 ◽  
Author(s):  
Yanhua Wang ◽  
Fuying Ma ◽  
Eva L. Rodriguez ◽  
Janet D. Klein ◽  
Jeff M. Sands

Aldosterone indirectly regulates water reabsorption in the distal tubule by regulating sodium reabsorption. However, the direct effect of aldosterone on vasopressin-regulated water and urea permeability in the rat inner medullary collecting duct (IMCD) has not been tested. We investigated whether aldosterone regulates osmotic water permeability in isolated perfused rat IMCDs. Adding aldosterone (500 nM) to the bath significantly decreased osmotic water permeability in the presence of vasopressin (50 pM) in both male and female rat IMCDs. Aldosterone significantly decreased aquaporin-2 (AQP2) phosphorylation at S256 but did not change it at S261. Previous studies show that aldosterone can act both genomically and non-genomically. We tested the mechanism by which aldosterone attenuates osmotic water permeability. Blockade of gene transcription with actinomycin D did not reverse aldosterone-attenuated osmotic water permeability. In addition to AQP2, the urea transporter UT-A1 contributes to vasopressin-regulated urine concentrating ability. We tested aldosterone-regulated urea permeability in vasopressin-treated IMCDs. Blockade of gene transcription did not reverse aldosterone-attenuated urea permeability. In conclusion, aldosterone directly regulates water reabsorption through a non-genomic mechanism. Aldosterone-attenuated water reabsorption may be related to decreased trafficking of AQP2 to the plasma membrane. There may be a sex difference apparent in the inhibitory effect of aldosterone on water reabsorption in the inner medullary collecting duct. This study is the first to show a direct effect of aldosterone to inhibit vasopressin-stimulated osmotic water permeability and urea permeability in perfused rat IMCDs.


1986 ◽  
Vol 250 (3) ◽  
pp. R341-R347 ◽  
Author(s):  
T. Miwa ◽  
H. Nishimura

The mammalian-type nephrons of avian kidneys contain a Henle's loop that runs parallel to the collecting ducts and the vasa recta. Thus we examined whether the thick ascending limb (TAL) of Henle's loop of the avian kidney acts as a diluting segment by measuring water and Cl transport in the isolated and perfused TAL of the quail, Coturnix coturnix. The TAL showed a lumen-positive transepithelial voltage (Vt) (+9.4 +/- 0.4 mV, n = 28). Net water flux (Jv) was nearly zero when the TAL was perfused and bathed with isosmotic solution. When the osmotic gradient was imposed, Jv increased only slightly, and thus the osmotic water permeability (Lp) was low. Arginine vasotocin (AVT) added to the hyperosmotic bath did not alter either Jv, Lp, or Vt. Cl efflux (lumen to bath, 370.4 +/- 27.7 peq X mm-1 X min-1) was higher than Cl influx (bath to lumen, 98.6 +/- 14.3 peq X mm-1 X min-1) when measured in the different tubules. AVT showed no effect on Cl efflux. These results indicate that in the TAL of the quail osmotic water permeability is low while net Cl reabsorption is present, suggesting that the TAL functions as a diluting segment.


1987 ◽  
Vol 253 (3) ◽  
pp. F563-F575
Author(s):  
J. C. Williams ◽  
J. A. Schafer

A computer model of the proximal tubule of the rabbit is described in which the tubule is treated as a single cylindrical barrier to the flow of solute and water between lumen and bath, and volume absorption is assumed to be driven exclusively by hydrostatic and osmotic pressure differences across this barrier. The model mimics the function of the tubule in two in vitro preparations: in simulations of the isolated tubule perfused under oil, the model correctly describes the solute concentration gradients that exist between the perfusate and absorbate and predicts differences in solute concentrations among absorbate droplets on the same tubule if luminal concentration becomes limiting. This prediction was tested experimentally with glucose and found to be correct. In simulations of the tubule perfused in an aqueous bath, the role of transmural hydrostatic pressure was explored; it is predicted that, at normal rates of in vitro perfusion (approximately 10 nl/min), increases in pressure have very little effect on volume absorption but can greatly alter the osmotic differences present across the wall of the tubule, especially with high values of osmotic water permeability. At high rates of perfusion, the ability of the tubule to produce a lumen hypotonic to the bath is reduced, but the direct effects of pressure on volume absorption become more apparent, resulting in relatively little effect of perfusion rate on volume absorption if the osmotic water permeability is sufficiently high. A similar relationship was seen experimentally. In all, this simple model provides a good prediction of function in isolated perfused tubules without any assumptions of hypertonic compartments within the epithelium.


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