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Functional Anatomy of the Kidney

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Abstract

The kidney is one of the two major excretory organs in the body. It is involved in numerous vital bodily functions, including regulation of acid-base balance; maintenance of blood pressure via regulation of sodium excretion; maintenance of plasma osmolality via regulation of water excretion; regulation of plasma potassium, calcium, and phosphorus; elaboration of erythropoietin to maintain hematocrit; and hydroxylation of vitamin D. This chapter provides an overview of the functional anatomy of the kidney, with an emphasis upon the major ultrastructural and functional features of each nephron segment in order to provide an anatomic and functional framework for understanding renal function and toxicology. The kidney is composed of a variety of highly specialized nephron segments that are composed of distinct epithelial cell types, which engender specific transport properties to nephron segments where they reside. The kidney is uniquely sensitive to toxins and toxicants because they receive and filter an enormous quantity of blood.

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Previous studies in the mammalian proximal tubule have suggested that para-aminohippurate (PAH) secretion is approximately threefold greater in the straight segment, or pars recta, than in the convoluted segment, or pars convoluta. However, the possibility that the site of maximal PAH secretion might be related better to particular tubule segments as identified by cell type had not been explored. In addition, the presence or absence of differences in PAH secretion between morphologically identical regions of superficial (SF) vs. juxtamedullary (JM) proximal tubules has not been examined. These issues were studied using a combination of histologic methods and measurement of [(3)H]PAH secretion in isolated perfused tubules. Measurements of microdissected SF and JM proximal tubules from young and adult rabbits revealed that SF proximal tubules were slightly but significantly longer than JM tubules ([young rabbits: SF, 8.69+/-SE 0.14 mm vs. JM, 7.97+/-SE 0.13 mm; P < 0.01] [adult rabbits: SF, 10.61+/-SE 0.28 mm; JM, 9.17+/-SE 0.19 mm; P < 0.001]). Light and electron microscopy revealed three sequential segments (S(1), S(2), and S(3)) along the length of SF and JM proximal tubules as defined by cell type. PAH secretion was measured in each of these three segments by the isolated perfused tubule technique. Net PAH secretion in fmol/mm per min in SF proximal tubules was: S(1), 281+/-SE 21; S(2), 1,508+/-SE 104; S(3), 318+/-SE 46. Corresponding values in JM proximal tubules were 353+/-SE 31, 1,391+/-SE 72, and 188+/-SE 23. Net PAH secretion did not differ between comparable segments of SF and JM proximal tubules. It is concluded that differences in PAH secretion along the proximal tubule correlate best with cell type rather than the arbitrary division of the proximal tubule into pars convoluta and pars recta according to its external configuration. Evidence of functional heterogeneity between comparable segments of SF and JM proximal tubules was not observed.
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A circuit is described that can produce a high output voltage to automatically short-circuit an epithelial tissue mounted in an Ussing chamber. Because of its high output voltage it can short-circuit preparations requiring over 500 muA even in conjunction with high-resistance agar bridges. The circuit, which is easy to build, uses an inexpensive, low-voltage, integrated circuit operational amplifier, which is electrically isolated from the high-voltage part of the circuit by an optical isolator. The device can also be modified for use as a high impedance preamplifier for monitoring the spontaneous membrane potential.
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Studies utilizing in vitro microperfusion were designed to examine whether urea is actively or passively transported across superficial and juxtamedullary straight segments of rabbit proximal tubules. With perfusate and bath solutions containing 1 mM urea and electrolytes similar to normal plasma, the efflux (lumen-to-bath) isotopic permeability (X 10(-5) cm s-1) of superficial segments was 1.37 +/- 0.16 and of juxtamedullary segments was 2.14 +/- 0.20. In the same tubules, the influx (bath-to-lumen) isotopic permeability was 3.70 +/- 0.35 in superficial segments and 4.75 +/- 0.37 in juxtamedullary segments. Despite net water movement in the opposite direction (0.5 nl mm-1 min-1), the influx rate was significantly higher than the efflux rate of urea in both groups. With a low perfusion rate (2 nl/min) and equivalent specific activities of [14C]urea in bath and perfusate, the collected-to-perfused ratio of [14C]urea, corrected for volume marker change, was 1.07 +/- 0.01 in superficial and 1.09 +/- 0.01 in juxtamedullary nephrons, thus indicating net secretion in both segments. In separate studies urea influx was inhibited by hypothermia (decrease from 37 degrees to 28 degrees C), by phloretin (0.1 mM in bath), by cyanide (1 mM), but not by probenecid (0.2 mM). In each case the inhibition was highly significant and reversible. These data suggest that urea is actively secreted by the straight segments of both the superficial and juxtamedullary proximal tubules. These segments may, therefore, contribute significantly to the high urea concentration found at the bend of Henle's loop by micropuncture.
Article
Water rapidly crosses the plasma membrane of red blood cells (RBCs) and renal tubules through specialized channels. Although selective for water, the molecular structure of these channels is unknown. The CHIP28 protein is an abundant integral membrane protein in mammalian RBCs and renal proximal tubules and belongs to a family of membrane proteins with unknown functions. Oocytes from Xenopus laevis microinjected with in vitro-transcribed CHIP28 RNA exhibited increased osmotic water permeability; this was reversibly inhibited by mercuric chloride, a known inhibitor of water channels. Therefore it is likely that CHIP28 is a functional unit of membrane water channels.
Article
The thin limb segments of the long loop of Henle are thought to play important roles in the urinary concentrating mechanism. In this study, we present new approaches to the identification, dissection, and in vitro perfusion of individual thin limb segments from all levels of the chinchilla renal medulla, including the deepest portions of the papilla. We have applied these techniques to the investigation of the osmotic water permeability along the chinchilla long loop of Henle. The results demonstrate that the osmotic water permeability of the thin descending limb is not uniformly high along its length, as previously thought, but that the distal 20% of the long-loop descending limb has a very low water permeability (approximately 50 microns/s). The transition to the low water permeability region of the thin descending limb is accompanied by a relatively abrupt change in morphology (increased cellularity and decreased diameter) that is readily perceptible in the perfused segments and even in the dissection dish. In contrast, the upper part of the chinchilla long-loop thin descending limb had an extremely high osmotic water permeability (greater than 2,000 microns/s) as observed in other species. Thin ascending limbs from deep in the inner medulla had water permeabilities that were indistinguishable from zero, as previously found in thin ascending limbs from near the inner-outer medullary junction. The presence of a low-water-permeability portion of the long-loop thin descending limb in chinchilla may have important implications with regard to the inner medullary concentrating process. A relatively low osmotic water permeability (397 microns/s) was also found in the deep inner medullary portion of the thin descending limb from the rat.
Article
We have shown that urea transport across the terminal inner medullary collecting duct (terminal IMCD) is mediated by a vasopressin-stimulated, facilitated diffusion process exhibiting properties consistent with a transporter. To investigate whether hypertonic NaCl, as exists in vivo in the inner medulla, affects urea permeability, we studied isolated perfused rat terminal IMCD segments. Perfusate and bath osmolality were varied symmetrically by adding or removing NaCl or mannitol. Urea permeability rose progressively when osmolality was increased with NaCl or mannitol from 290 to 690 mOsm/kg H2O in the absence of vasopressin; there was no further increase at 890 mOsm/kg H2O. In the presence of 10(-8) M arginine vasopressin, urea permeability increased when NaCl was added to raise osmolality from 290 to 490 mOsm/kg H2O but there was no further increase at 690 mOsm/kg H2O. When 1 mM 8-bromo cyclic AMP was added to the bath, raising NaCl still increased urea permeability. These results suggest that urea transport across the rat terminal IMCD is regulated both by vasopressin and by osmolality at values present in the renal inner medulla. Osmolality seems to activate urea transport across the rat terminal IMCD by mechanisms distinct from those of vasopressin or cyclic AMP.
Article
Currents through individual Na channels in the apical membrane of the rat cortical collecting tubule were resolved by using the patch-clamp technique. In cell-attached patches, the channels had a conductance of 5 pS with 140 mM NaCl in the pipet. The conductance was a saturable function of external Na, with a maximal value of about 8 pS and a half saturation at about 75 mM Na. In excised inside-out patches, the selectivity of the channels for Na over K was estimated from reversal potentials to be at least 10:1. The channels underwent spontaneous transitions between open and closed states. Both states had mean lifetimes of 3-4 sec. Amiloride (0.5 microM) added to the pipet induced more frequent closures and openings of the channels and a reduction in the mean open time. These channels are presumed to mediate Na reabsorption by this nephron segment in vivo.
Article
We examined the hypothesis that proton-potassium-activated adenosine triphosphatase (H-K-ATPase) mediates K absorption and acidification in the inner stripe of the outer medullary collecting duct (OMCDi). Rabbits were fed a low-K diet (0.55% K) for 7-14 d because we have demonstrated previously that this low-K diet stimulates K-absorptive flux by the OMCDi. Proton secretion was measured as net total CO2 flux (JTCO2) by microcalorimetry. After basal collections, either vehicle or an inhibitor of gastric H-K-ATPase, omeprazole (0.1 mM), was added to the perfusate during the second period. Addition of vehicle to the perfusate changed neither the transepithelial voltage (VT, in millivolts) nor the JTCO2. In contrast, the addition of omeprazole (0.1 mM) to the perfusate abolished JTCO2 (from 14.5 +/- 5.6 to -0.1 +/- 3.1 pmol.mm-1.min-1) without significantly affecting VT. In additional experiments, in 16 tubules there was significant net K absorption (JK) of 5.0 +/- 1.0 pmol.mm-1.min-1 during the basal period, which exceeded the rate of K absorption that could be attributed to a paracellular voltage-mediated pathway (JKP = 1.0 +/- 0.4 pmol.mm-1.min-1, P less than 0.01). Administration of vehicle did not significantly affect either VT or JK. However, omeprazole abolished JK (from 5.1 +/- 1.0 to 0.1 +/- 2.5 pmol.mm-1.min-1) without affecting VT or JNa. The present results demonstrate that the OMCDi possesses an active, omeprazole-sensitive acidification and K-absorptive mechanism. These findings are consistent with the presence of H-K-ATPase activity in this nephron segment.
Article
Synthesis of the sodium pump, Na+-K+-ATPase, is regulated by thyroid hormone in responsive tissues. The purpose of this study was to determine if triiodothyronine (T3) regulates the concentration of the mRNAs coding for the two enzyme subunits, alpha and beta, and the time course of the response. A single dose of T3 (250 micrograms/100 g body wt) was administered to hypothyroid rats that were killed at various times after injection. In the kidney cortexes of the T3-injected animals, as well as hypothyroid and euthyroid rats, alpha- and beta-mRNA concentrations were measured by dot blot using cDNAs corresponding to the two mRNAs; alpha-subunit abundance was measured by Western blot using antibodies to the enzyme, and Na+-K+-ATPase activity was measured enzymatically. alpha- and beta-mRNAs increased coordinately, after a 6-h time lag to 1.6-fold over hypothyroid levels by 12 h after T3. alpha-Subunit abundance increased significantly by 48 h and to 1.4-fold over hypothyroid by 72 h after T3. Na+-K+-ATPase activity increased with the same time course as the increase in alpha-subunit abundance to 1.3-fold over hypothyroid by 72 h after T3. We conclude that T3 regulates Na+-K+-ATPase synthesis and activity by coordinately increasing the mRNAs of both the alpha- and beta-subunits of the enzyme.
Article
To determine the exact site and mechanism of action of thiazide diuretics, effects of 10(-4) M trichlormethiazide (TCM) on NaCl transport were examined in the distal convoluted tubule (DCT), the connecting tubule (CNT) and the cortical collecting duct (CCD) of rabbit kidney by the in vitro microperfusion technique. TCM added to the lumen decreased lumen-to-bath 36Cl flux (JCl(LB)) only in the CNT without changing the transmural voltage (VT). In the DCT, 10(-4) M furosemide did not change JCl(LB) even if it was added to the lumen with 10(-4) M TCM, whereas 10(-5) M amiloride in the lumen decreased the lumen-to-bath 22Na flux (JNa(LB)) and VT. In the CNT, TCM added to the lumen did not affect the bath-to-lumen 36Cl flux. Addition of TCM to the bath slightly decreased JCl(LB). Luminal addition of 10(-4) M TCM also decreased JNa(LB). Amiloride at 10(-5) M in the lumen decreased both JNa(LB) and VT. Addition of TCM with 10(-5) M amiloride further decreased JNa(LB) without affecting VT, indicating that TCM affects the electroneutral Na+ transport, which is distinct from the amiloride-sensitive conductive Na+ pathway. When Na+ was removed from the lumen, JCl(LB) was markedly decreased, but addition of TCM did not cause further decrease in JCl(LB). Furosemide did not affect JCl(LB), but addition of both 10(-4) M TCM and furosemide decreased JCl(LB), indicating that Na+-K+-2Cl- cotransport is not involved in the action of TCM. Removal of HCO3- slightly decreased JCl(LB), and TCM caused further decrease in JCl(LB). Amiloride at 10(-3) M, a concentration supposed to inhibit the Na+/H+ antiport, slightly decreased JCl(LB), and addition of TCM caused a further marked decrease in JJl(LB). The similar results were also obtained when the combined effects of 10(-3) M 4,4'-diisothiocyano-stilben-2,2'-disulfonate(DIDS) and 10(-4) M TCM were examined. These findings suggest that the parallel antiport of Na+/H+ and Cl-/HCO3- is not involved in the action of TCM. By excluding other possible mechanisms involving neutral Na+-dependent Cl- transport, we conclude that TCM inhibits Na+-Cl- cotransport in the luminal membrane of the rabbit CNT.
Article
In a review published in 1977, Sachs [1] concluded that “biochemical studies on renal tissue have so far not shown the presence of any active pump mechanism other than Na,K-ATPase,” and pointed out that transport of Na, K, H, Cl, and HCO3can be explained by various modes of direct or indirect coupling to Na,K-ATPase. In the intervening years a large body of information further established the crucial role of Na,K-ATPase in tubular electrolyte transport, while at the same time providing evidence for the presence of other transport ATPases in the kidney. In parallel, microdissection and microanalysis techniques have made possible the examination of metabolic and enzymatic processes in discrete tubule segments that revealed the remarkable biochemical heterogeneity of the nephron [2], much as in the recent past in vitro tubule microperfusion has demonstrated the axial heterogeneity of its transport processes [3, 4]. Given the functional specialization of different nephron segments, one can easily appreciate why studies of renal biochemistry with classical methods using tissue slices or homogenates are subject to the same kind of limitations as, for example, the analysis of kidney function with clearance techniques: Both provide information about the kidney as a whole, rather than about its component units. To understand nephron function, one has to study it in its subdivisions of interest; to evaluate the regulation and the role of ATPases in tubular transport, it is essential to examine the behavior of these enzymes in individual nephron segments, and first to delineate their location along the nephron. This paper reviews the nephron distribution and function of several classes of ATPase, with emphasis on the sodium- and potassium-activated adenosine triphosphatase (Na,K-ATPase), and including the calcium- and proton-ATPases, about which much less is known at this time.
Article
Mineralocorticoid hormones stimulate Na+ absorption and K+ secretion by the cortical collecting tubule. There is good evidence that this stimulation involves increasing luminal membrane Na+ permeability and the turnover rate (or number) of the Na+-K+ pumps. These experiments were designed to examine whether mineralocorticoid hormones also increase cell K+ permeability. Using 42K tracer measurements in tubules treated with amiloride to inhibit active Na+ and K+ transport, passive K+ permeation increased with increasing mineralocorticoid effect. Net Na+ absorption and the (passive) K+ efflux rate coefficient (KK) showed a linear relationship. The stimulatory effect was evident in vitro since 0.2 microM aldosterone added to the bath of tubules harvested from NaCl-loaded rabbits increased KK at 3 hrs while time controls showed no change. Since these tubules were also treated with amiloride, this increase in KK was not dependent on increasing Na+ absorption. The results indicate that in addition to the well-described effects of aldosterone on Na+ permeability and cell metabolism, the mineralcorticoid effect includes an increase in cellular K+ permeability.
Article
To test prolactin (PRL) action on osmoregulation in mammals, we evaluated in the rat the effect of this hormone on a major enzyme in renal regulation of water and electrolyte: renal Na-K-ATPase. Enzyme activity was determined by cytochemistry in medullary ascending limb (MAL) and distal convoluted tubule (DCT) from rats treated either by bromocriptine, or by PRL. Three hours after a bromocriptine injection (0.1 mg/100 g IP) a significant decrease of Na-K-ATPase activity is observed in both MAL (80% of control values, p less than 0.001) and DCT (78% p less than 0.01). Reciprocally, a significant (p less than 0.001) increase in enzyme activity is induced 3 h after a single PRL injection (140 micrograms/100 g IM), in both segments (MAL: 165%, DCT: 172% of control activities) and persists 6 h after the injection (MAL: 130%, DCT: 118%). Na-K-ATPase activity was correlated to plasma PRL levels (r = 0.78 in DCT, r = 0.89 in MAL). A direct effect of PRL on the tubule is suggested by results from experiments in which PRL, at various concentrations, is added in vitro on renal slices before Na-K-ATPase activity measurements. The increase in Na-K-ATPase activity exhibits a log-dose dependency with PRL concentration (p less than 0.01) and is still observed when AVP antagonist is added before PRL incubation, ruling out the possible role of AVP contamination of PRL. These results suggest a direct effect of PRL on renal Na-K-ATPase in MAL and DCT.
Article
Our previous studies in cortical collecting ducts isolated from rat kidneys have shown that vasopressin increases both sodium absorption and potassium secretion, while bradykinin inhibits sodium absorption without affecting potassium transport. To determine which anions are affected by these agents, we perfused cortical collecting ducts from rats treated with deoxycorticosterone and measured net chloride flux, net bicarbonate flux (measured as total CO2), transepithelial voltage, and the rate of fluid absorption. Arginine vasopressin (10(-10) M in the peritubular bath) caused a sustained sixfold increase in net chloride absorption and a two- to threefold increase in the magnitude of the lumen negative transepithelial voltage. Before addition of vasopressin, the tubules secreted bicarbonate. Vasopressin abolished the bicarbonate secretion, resulting in net bicarbonate absorption (presumably due to proton secretion) in many tubules. Bradykinin (10(-9) M added to the peritubular bath) caused a reversible 40% inhibition of net chloride absorption, but did not affect the transepithelial voltage or the bicarbonate flux. We concluded: (a) that arginine vasopressin stimulates absorption of chloride and inhibits bicarbonate secretion (or stimulates proton secretion) in the rat cortical collecting duct; and (b) that bradykinin inhibits net chloride absorption in the rat cortical collecting duct without affecting transepithelial voltage or bicarbonate flux. Combining these results with the previous observations on cation fluxes described above, we conclude that bradykinin inhibits electroneutral NaCl absorption (or stimulates electroneutral NaCl secretion) in the rat cortical collecting duct.
Article
In order to further characterize Cl- transport of the thin ascending limb of Henle's loop (TAL), we observed the effects of glutaraldehyde on Na+ and Cl- transport in hamster TAL perfused in vitro. We found that 0.1 mol/l glutaraldehyde added either to the lumen or to the bath caused a rapid irreversible reversal of the NaCl diffusion potential. This was mainly accounted for by an inhibition of Cl- permeability (10(-7) cm2 s-1) from 93.51 +/- 8.39 to 14.89 +/- 3.91 (P less than 0.01, n = 9). By contrast, Na+ permeability changed little from 34.18 +/- 3.27 to 26.56 +/- 2.74 (P less than 0.01, n = 6). Glutaraldehyde treatment abolished the halogen-permselectivity of the TAL as determined by the voltage deflection seen upon ionic substitution. Permeabilities for Cl-, Br-, I-, and SCN- relative to Na+ were changed from 3.16 +/- 0.20, 3.22 +/- 0.19, 2.97 +/- 0.26 and 4.36 +/- 0.36 to 0.38 +/- 0.07, 0.35 +/- 0.06, 0.36 +/- 0.07 and 0.58 +/- 0.05, respectively. The effect of glutaraldehyde on the NaCl diffusion potential was dose-dependent in the range from 10(-5) to 10(-1) M. The effect was reversible at concentrations lower than 10(-3) M. Glutaraldehyde did not affect the NaCl diffusion potential of the long-loop descending limb. These observations constitute additional evidence that the mechanism of Cl- transport across the TAL is different from that of Na+ transport. Glutaraldehyde might inhibit Cl- transport in the TAL by cross-linking amino acid residues of the proteins essential for halogen transport across this segment.
Article
The stromal-to-tear transport of Cl- by the corneal epithelium of the frog is increased by pharmacological effectors (secretagogues) that are known to raise the intracellular levels of cyclic AMP or Ca2+. It has been shown in the past that the Cl- secretagogues increase the apical membrane permeability to Cl- and thus facilitate the cell-to-tear flux of the anion. In this report, we combine transepithelial and microelectrode studies to show that three of these secretagogues, epinephrine, the Ca2+ ionophore A23187, and forskolin, also increase the K+ conductance of the basolateral membrane by two- to threefold. The increase in the K+ conductance is not dependent on membrane potential, since this increase occurred equally when the basolateral membrane potential either exceeded 60 mV, as measured with microelectrodes, or was depolarized by voltage clamping after apical permeabilization with amphotericin B. It is proposed that both Cl- and K+ conductances are under the control of intracellular mediators that act independently on each pathway. The increase in basolateral K+ conductance favors the Cl- secretory process.
Article
The possible role of protein kinase c in regulating the electrical events in the B-cell plasma membrane was examined by using the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA), a known activator of this enzyme. TPA has been found to enhance glucose- and sulfonylurea-induced insulin secretion with little or no effect on the fluxes of 86Rb+ or 45Ca2+ across the plasma membrane. TPA, 0.2 microM, did not influence the membrane potential from 0 to 5.6 mM glucose but increased by two- to threefold the fraction of the plateau phase of the oscillatory electrical activity induced by 7.0-11.1 mM glucose. This effect of TPA was completely blocked by 0.5 mM spermidine, an inhibitor of protein kinase c. However, spermidine had no influence on the electrical activity elicited by glucose alone. Glyburide, 10 nM, initiated slow depolarization and constant spike activity after about 18 and 25 min, respectively. TPA or 2.8 mM glucose reduced the lag period for glyburide to elicit an electrical response by about 75%. The duration of the spikes was increased two- to threefold by the presence of glucose or TPA with glyburide. There were also characteristic differences in the shape of the spikes under each experimental condition. Spermidine inhibited the influence of glucose, but not TPA, on the glyburide-induced electrical response. These results indicate that TPA may influence stimulant-induced electrical events via protein kinase c or by directly altering the ionic permeability of the plasma membrane.
Article
To demonstrate that osmotic work can be accomplished across the inner medullary collecting duct (IMCD) by the difference in reflection coefficients for urea and NaCl, phenomenological coefficients for urea and NaCl transport were determined in isolated segments of the hamster IMCD perfused in vitro. Arginine vasopressin at 100 microU/ml increased urea permeability from 11.5 +/- 2.9 to 31.7 +/- 4.2 x 10(-7) cm2 s-1 in the middle IMCD but not in the upper IMCD. Urea transport in the middle IMCD consisted of two components, transport with saturable kinetics and simple passive diffusion. Permeability to Na+ was very low (2 x 10(-7) cm2 s-1). Reflection coefficients as measured by the equiosmolality method, with raffinose being a reference solute, were 0.87 +/- 0.05 and 0.71 +/- 0.04 for urea and 1.03 +/- 0.07 and 0.91 +/- 0.04 for NaCl in the upper and the middle IMCD, respectively. Reflection coefficient for urea in the middle IMCD was 0.68 when determined by the zero volume flux method. When the middle IMCD was perfused with bicarbonate Krebs-Ringer (BKR) solution containing 200 mmol/l urea, the replacement of urea in the bathing fluid with equisomolal NaCl caused large volume flux (3.81 +/- 0.45 nl mm-1 min-1) associated with dilatation of intercellular space. The existence of vasopressin in the bath was essential for this phenomenon. This effect was inhibited by 5 x 10(-4) M phloretin in the bath, suggesting that the vasoressin-stimulated urea transport is responsible for this phenomenon. From these observations, we conclude that transport parameters of the middle IMCD are appropriate for accomplishment of osmotic work across this segment in the absence of physicochemical osmotic gradients.
Article
The interaction of 12-HETE with MDCK cells was investigated to determine whether this lipoxygenase product might influence tubular epithelial function. MDCK cells took up 12-HETE from both the luminal and interstitial surfaces. Much of the 12-HETE was incorporated into phospholipids but, unlike arachidonic acid, there was no specificity for the sn-2 position. 12-HETE was incorporated without chemical modification, suggesting that it might perturb tight packing in the lipid bilayer. 12-HETE uptake was reduced by the presence of arachidonic acid, but not by oleic acid. 12-HETE was removed from the cell lipids more rapidly than arachidonic acid and it was released into the medium in the form of both more and less polar metabolites. When MDCK cells were incubated with 12-HETE, their capacity to produce prostaglandin E2 was reduced. Since 12-HETE enters epithelial phospholipids and is converted to metabolic products and reduces prostaglandin production, it apparently has the capacity to modulate renal function if it is released in the proximity of the tubular epithelium.
Article
The existence of a steep osmotic gradient in the renal medullary interstitium is the most critical in the formation of concentrated urine [1]. The architectural organization of the renal tubules and blood vessels in the medulla constitutes counterflow systems which are essential for both generating and maintaining high osmotic pressure of the renal medulla [2–4]. While it has been generally accepted that active NaCl transport in the thick ascending limb of Henle's loop plays the most fundamental role in the operation of the countercurrent multiplication system in the renal medulla, it is still a matter of considerable dispute whether the thin ascending limb (tAL) also has an active salt transport system to provide a “single effect” necessary for the operation of the countercurrent multiplication system.
Article
We have analyzed the histotopography of the renal medulla of the rat in terms of cycles and separations. Cycles are pathways by which solute leaving the medulla in an ascending structure (AVR, AHL) is returned to a deeper medullary level. Separations are based on spatial incontiguity and special characteristics of the interstitium and blood supply. The two concepts are complementary: the compartmentalization resulting from separations imparts specificity to the cycles. Structural lateral heterogeneity, consisting in distinct domains organized around vascular bundles, is present in one form or another in all three medullary zones. Such compartmentalization probably leads to heterogeneity in interstitial solute concentrations, a state of affairs inconsistent with the requirements of a "central core". In all such considerations of exchanges between compartments, the lack of a unitary interstitium must be borne in mind. Instead, three general types of interstitium may be distinguished: corresponding roughly to those of the OS and VB, the interbundle region of the IS, and the IM. Among the histotopographic features of the renal medulla not usually included in models of the urinary concentrating mechanism but likely to have functional significance are the association of CD with completely distinct populations of AVR and AHL in the OM and IM; a clear-cut separation throughout the medulla between cycles involving long loops and those involving short loops; the lack of an effective countercurrent association between ascending and descending limbs of short loops in the IS; and a pronounced separation of the venous drainage of the IM from that of the OM.
Article
There is increasing evidence of acidification along the entire mammalian collecting duct including the inner medullary collecting duct (IMCD). Recent studies have provided morphologic evidence that the intercalated cells are involved in hydrogen ion secretion in the cortical and outer medullary collecting duct of the rat. In the present study we performed a quantitative and qualitative morphologic examination of the intercalated cells in the IMCD of the rat and compared the results to observations obtained from intercalated cells in the collecting duct in the inner stripe of the outer medulla (OMCDi). Kidneys of male rats were preserved by in vivo perfusion with glutaraldehyde and processed for morphologic evaluation. With light microscopy and scanning electron microscopy intercalated cells were found in the outer third of the IMCD (IMCD1) and accounted for 10% of the total cell population. They were absent in the terminal two-thirds of the IMCD. Examination of the intercalated cells using transmission electron microscopy revealed striking similarities between the cells of the IMCD1 and those in the OMCDi. In addition, no differences were found in the surface densities of the apical or basolateral plasma membranes or the volume densities of the mitochondria of the intercalated cells in the two regions. In light of the morphologic similarity with the intercalated cells of the OMCDi that are believed to be involved in hydrogen ion secretion, it is likely that the intercalated cells of the IMCD1 are also involved in the acidification of tubular fluid.
Article
To compare passive urea transport across the inner medullary collecting ducts (IMCDs) and the papillary surface epithelium (PSE) of the kidney, two determinants of passive transport were measured, namely permeability coefficient and surface area. Urea permeability was measured in isolated perfused IMCDs dissected from carefully localized sites along the inner medullas of rats and rabbits. Mean permeability coefficients (X 10(-5) cm/s) in rat IMCDs were: outer third of inner medulla (IMCD1), 1.6 +/- 0.5; middle third (IMCD2), 46.6 +/- 10.5; and inner third (IMCD3), 39.1 +/- 3.6. Mean permeability coefficients in rabbit IMCDs were: IMCD1, 1.2 +/- 0.1; IMCD2, 11.6 +/- 2.8; and IMCD3, 13.1 +/- 1.8. The rabbit PSE was dissected free from the underlying renal inner medulla and was mounted in a specially designed chamber to measure its permeability to urea. The mean value was 1 X 10(-5) cm/s both in the absence and presence of vasopressin (10 nM). Morphometry of renal papillary cross sections revealed that the total surface area of IMCDs exceeds the total area of the PSE by 10-fold in the rat and threefold in the rabbit. We conclude: the IMCD displays axial heterogeneity with respect to urea permeability, with a high permeability only in its distal two-thirds; and because the urea permeability and surface area of the PSE are relatively small, passive transport across it is unlikely to be a major source of urea to the inner medullary interstitium.
Article
The protein A-gold technique was used to localize carbonic anhydrase isozymes on ultrathin frozen sections of kidney collecting duct epithelial cells and erythrocytes. The particulate nature of the gold marker gives a more precise appreciation of the intracellular distribution of this enzyme than has been previously possible, and allows the intensity of the labeling to be quantified. Intercalated cells showed four times more labeling over the cytosol than adjacent principal cells in collecting ducts from the inner stripe of the outer medulla: by double-labeling using protein A-gold particles of different sizes, carbonic anhydrase isozymes B and C were simultaneously localized in erythrocytes.
Article
Several factors interact to maintain precise control of electrolyte transport in the mammalian cortical collecting duct. We have studied the effects of deoxycorticosterone, arginine vasopressin, and bradykinin on net transepithelial sodium and potassium transport in isolated, perfused rat cortical collecting ducts. Chronic administration of deoxycorticosterone to rats increased both sodium absorption and potassium secretion above very low basal levels. Consequently, deoxycorticosterone-treated rats were used for all remaining studies. Arginine vasopressin (10(-10) M in the bath) caused a sustained fourfold increase in net sodium absorption and a sustained threefold increase in net potassium secretion. Bradykinin (10(-9) M in the bath) caused a reversible 40-50% inhibition of net sodium absorption without affecting net potassium transport or the transepithelial potential difference. In the perfusate, up to 10(-6) M bradykinin had no effect. We conclude: As in rabbits, chronic deoxycorticosterone administration to rats increases sodium absorption and potassium secretion in cortical collecting ducts perfused in vitro. Arginine vasopressin causes a reversible increase in net potassium secretion and net sodium absorption. Bradykinin in the peritubular bathing solution reversibly inhibits net sodium absorption, possibly by affecting an electroneutral sodium transport pathway.
Article
Structural differences in thin limbs of Henle: Physiological implications. Rat thin limbs of Henle (TL) were studied by electron microscopy. Descending and ascending thin limbs (DTL, ATL) were identified by tracing their continuity with known structures and by histotopography. Two types of TL were identified. Type I showed cellular interdigitation and very shallow intercellular occluding junctions. Junctions between noninterdigitating cells in type II had longer apical-basal depths. In short loops, DTL were type II. In long loops, DTL were type 1 in the outer medulla and possessed “complex” features such as microvilli and basal cytoplasmic invaginations. “Complex” type I DTL extended to varying distances in the inner medulla, where they underwent transition to type II limbs. ATL were type I, but appeared “simple” compared to the upper portions of DTL (no microvilli or basal invaginations). Transition from type II to “simple” type I epithelium occurred in the terminal portions of DTL at short distances (56 to 133 µ) before their junctions with ATL at the bends. Counts showed that profiles of “complex” type I DTL were outnumbered by type II DTL in the upper one-half, and virtually absent in the lower one-half of the inner medulla. Ultrastructural studies suggest that the degree of passive ion permeation across epithelia may be determined by the depth of their occluding junctions. Thus, type I cell junctions may be more permeable than type II. Cellular interdigitation provides for a larger paracellular pathway in type I limbs, and thus may enhance their permeability. These considerations suggest a role for TL structural differences in medullary counter-current mechanisms.
Article
Permeability of medullary nephron segments to urea and water: Effect of vasopressin. High papillary urea concentrations are necessary for the formation of maximally concentrated urine, while low papillary urea concentrations are associated with less concentrated urine. In the present studies we examined the membrane characteristics that are important in determining the medullary urea concentration profiles. Using isolated segments of rabbit nephrons perfused in vitro, we measured the diffusional urea and water permeability in the medullary thick ascending limb of Henle (MTALH), the outer medullary collecting duct (MCD) and the papillary collecting duct (PCD), in the presence and absence of antidiuretic hormone (ADH). Segments of MTALH and MCD were essentially impermeant to 14C-urea. This degree of impermeability was maintained both with or without ADH. The urea permeability (× 10−5 cm sec−1) of the PCD was moderate (2.2± 0.3 without ADH, 2.4± 0.6 with ADH). Diffusional water permeability (× 10−5 cm sec−1) of the MTALH was low both with and without ADH; but the diffusional water permeability of the PCD increased from 39.9 ± 7.6 to 56.9 ± 8.6 (P < 0.002) with the addition of 200 µU/ml of ADH to the bath. In summary: 1) ADH has no influence on urea permeability across the nephron segments studied; 2) urea permeability of the PCD is higher than that of the MCD or the MTALH; and 3) ADH increases diffusional water permeability of the PCD but not of the MTALH. We conclude that pathways of urea movement are not the same as the principal pathways of water movement across the papillary collecting duct epithelium.
Article
Ultrastructure of the renal pelvic epithelium of the rat. The ultrastructure of the renal pelvis of six rats was examined. Transitional epithelium, similar to that found in the bladder, was limited to the contrapapillary surface and overlay cortex and outer medulla. Like the bladder, the luminal plasma membrane of these cells measured 125Å and its outer leaflet was thicker than the cytoplasmic. The papillary epithelium was comprised of simple cuboidal cells, whose luminal membrane was 75Å in thickness. In the fornix, groups of cells intermediate in appearance were observed with regions of both 75Å and 125Å membrane present along the luminal surface. A correlation was observed between the urea permeability of each pelvic cell type and the concentration of urea in the underlying renal parenchyma reported by other investigators ; that is, the urea-permeable papillary epithelial cells invest the inner medulla which is a region of high urea concentration, whereas the transitional cells believed to be impermeable overlie the urea-poor cortex and outer medulla. It is suggested that the pelvic epithelial cells may participate in the regulation of urea reabsorption from the pelvic urine.
Article
Countercurrent multiplication system without active transport in inner medulla. The present study reports a model for countercurrent multiplication in which both the descending limb of Henle (DLH) and thin ascending limb of Henle (ALH) operate as purely passive equilibrating segments. This model is based largely on transport characteristics obtained by perfusing isolated segments of rabbit nephrons in vitro. For this passive equilibration model to be operative the following segmental membrane characteristics should be present: thin DLH, relatively impermeant to solute but having high osmotic water permeability (osmotic water equilibration raises intraluminal Na concentration to higher levels than adjacent interstitium); thin ALH, more permeant to NaCl than urea and relatively impermeant to osmotic flow of water (hypoosmolal fluid can be generated passively by allowing more net NaCl to diffuse out than influx of urea); thick ALH, relativly water and solute impermeable with capacity of active outward NaCl transport (hypotonic fluid with a low NaCl concentration is developed); distal tubule, cortical and outer medullary collecting duct, impermeant to urea both in presence and absence of vasopressin (intraluminal urea is raised to high concentrations by virtue of water abstraction); inner medulla and papillary collecting duct, urea permeant (allows for generation of high interstitial urea concentration by passive diffusion down chemical concentration gradient). The salient feature of the proposed model is that the energy generated by active outward NaCl transport by thick ALH (expressed as high urea concentration in outer medullary CD by virtue of water abstraction) is transmitted to the papilla (by way of urea diffusing down its concentration gradient). In turn, papillary interstitial urea abstracts water out of DLH generating high intraluminal NaCl concentrations which allows the entire system to operate by passive diffusion of NaCl out of the thin ALH. By this model, the observed medullary concentration gradients are developed without invalidating the mass balance equations.
Article
Long loops of Henle in rat kidneys fixed during an osmotic diuresis were marked by microinjection with a silicone rubber. In histological serial sections corresponding descending and ascending thin limbs in the inner medullary zone were identified and their luminal diameters measured. At any level of the inner medulary zone, the luminal diameter of descending limbs is considerably smaller than that of corresponding ascending limbs. This diameter difference is most drastic at the border between inner and outer zones and diminishes towards the bend of the loops. Starting with a pre-bend enlargement the ascending thin limbs increase in diameter towards the outer medullary zone.
Article
Concentration of urine in a central core model of the renal counterflow system. In this model descending Henle's limbs (DHL), ascending Henle's limbs (AHL) and collecting ducts (CD) exchange with a central vascular core (VC) formed by vasa recta loops—assumed so highly permeable that the core functions as a single tube, open at the cortical end and closed at the papillary end. Solute supplied to the VC primarily by AHL increases VC osmolality and so extracts water from DHL and CD, increasing their osmolality while diluting AHL fluid. This single effect multiplied by the counterflow arrangement leads to a high papillary osmolality in all structures. Some of the solute may enter DHL to be recycled. In single solute system energy requirements for transport out of AHL decrease from outer to inner medulla. In two solute systems (e.g. salt and urea) mixing in the central core can supply part of the energy for the final concentration of urine. Urea cycling, regulated by ADH, allows active Na+ transport in the outer medulla and cortex to be used for concentration in the inner medulla.