Renal medullary circulation: morphological characteristics of vessels and their organization

The blood vessels of the renal medulla have several particular features. The supplying vessels of the renal medulla are the efferent arterioles of the juxtamedullary glomeruli. Thus, the blood supply is entirely postglomerular. The distribution of the blood within the renal medulla is effected by the descending vasa recta. Descending and ascending vasa recta form the vascular bundles. Descending vasa recta leave the bundles at any level of the medulla to feed the adjacent capillaries; the longest descending vasa recta reach the papillary tip. The capillary plexus are differently developed. A dense capillary plexus is present only in the inner stripe of the outer medulla. In the inner medulla the capillary plexuses are poorly developed. At these sites ascending recta contribute to the capillarisation. The ascending vasa recta originate from capillaries at any level of the medulla and ascend without joining together towards the intrarenal veins at the cortico-medullary border. They are capillary vessels with wide lumina. Within the outer stripe of the outer medulla these vessels are very narrowly associated with the tubules; interstitial spaces are very poorly developed at this site. These relationships are interpreted as a possible weak point of the medullary circulation in pathological situations.     

Ultrastructural differences between rat inner medullary descending and ascending vasa recta;

The ultrastructure of rat inner medullary vasa recta was studied by both conventional transmission and freeze-fracture electron microscopy. The identify of descending and ascending vasa recta in the inner medulla was established by tracing outer medullary descending vasa recta and ascending vasa recta into the inner medulla, as well as by the incomplete carbon labeling technique to identify isolated descending vessels or loops. As in the outer medulla, descending vessels possess thick continuous endothelium with pinocytotic vesicles budding off the luminal and basal plasma membranes (more numerous in the latter location), but no fenestrae. Ascending vasa show thin, attenuated endothelium with numerous (500 to 800 A fenestrae bridged by 40 A thick fenestral diaphragms. Intercellular junctions in both vessels are zonulae occludens with usually one, and uncommonly two or three zones of outer leaflet membrane fusion. Intramembranous particles are numerous in endothelial plasma membranes of descending vasa recta; similar particles are much fewer in corresponding ascending vasa recta endothelial plasma membranes. The ultrastructural differences between descending vasa recta and ascending vasa recta may reflect markedly different permeability properties, particularly to macromolecules, and may be relevant to recent functional studies on rat inner medullary vasa recta.     

A flavinogenic mutant of the yeast Pichia guilliermondii with impaired iron transport

We recently demonstrated that net fluid uptake occurs in the capillary system of the inner medulla. To define the site of fluid uptake, the concentration of protein was determined in plasma from descending vasa recta at the base and tip of the exposed papilla in Munich-Wister rats. The vasa recta plasma-to-arterial plasma protein concentration ratio (VR/P) was 1.43 +/- 0.09 at the base and 1.66 +/- 0.09 at the tip. These results, which indicate fluid loss from the descending vasa recta, are difficult to explain on the basic of hydraulic and oncotic forces alone. The osmolality of the contents of descending vasa recta increased between base and tip (delta = 72 +/- 30 mosmol/kg H2O). If the increase in osmolality of plasma in descending vasa recta lags behind that of the adjacent medullary interstitium, a transcapillary osmotic driving force exists favoring water loss from descending vessels. It is concluded that fluid uptake by the inner medullary circulation occurs beyond descending vasa recta in interconnecting capillaries or ascending vasa recta. In our view the most likely interpretation of these results is that fluid movement across vasa recta in the inner medulla is influenced by three forces: those owing to transcapillary differences in osmotic, oncotic, and hydraulic pressures.     

Impact of cyclo-oxygenase blockade on juxtamedullary microvascular responses to angiotensin II in rat kidney

1. Experiments were designed to evaluate the hypothesis that cyclo-oxygenase products modulate the influence of angiotensin II (AII) on the renal juxtamedullary microvasculature of enalaprilat-treated rats. 2. The in vitro blood-perfused juxtamedullary nephron technique was utilized to provide access to afferent arterioles, efferent arterioles and descending vasa recta located in the outer stripe of the outer medulla. 3. Baseline afferent arteriolar diameter was 20.8 +/- 1.9 microns in kidneys subjected to cyclo-oxygenase blockade (1 mumol/L piroxicam), a value significantly lower than that observed in untreated kidneys (26.1 +/- 1.0 microns). Baseline diameters of efferent arterioles and outer medullary descending vasa recta did not differ between untreated and piroxicam-treated groups. 4. Topical application of 1 nmol/L AII reduced blood flow through outer medullary descending vasa recta by 22 +/- 6% in untreated kidneys and by 24 +/- 7% in piroxicam-treated kidneys. 5. In untreated kidneys, AII (0.01-100 nmol/L) produced concentration-dependent afferent and efferent arteriolar constrictor responses of similar magnitudes. Neither afferent nor efferent arteriolar AII responsiveness was significantly altered in piroxicam-treated kidneys, although afferent responses exceeded efferent responses at AII concentrations > or = 10 nmol/L. 6. We conclude that endogenous cyclo-oxygenase products exert a vasodilator influence on juxtamedullary afferent arterioles under baseline conditions. Although cyclo-oxygenase inhibition had little effect on juxtamedullary microvascular responses to AII, the response to high AII concentrations may be modulated by cyclo-oxygenase products in a manner which delicately alters the relative influence of the peptide on pre- vs postglomerular resistances.     

Changes within vascular bundles of rodent kidneys caused by different diets

Morphological effects caused by two different diets (low protein-high water intake, and high protein-restricted water) on the vascular bundles in the outer medullary zone of the kidney were studied in the laboratory white mouse and in the golden spiny mouse (Acomys russatus, Muridae). In both rodents, when on a low protein-high water intake diet, considerable interstitial substance was found between the vasa recta of the bundles. No interstitial substance was found in animals on high protein-low water intake diet; as a result the vasa recta of the vascular bundle adhered closely. The low protein-high water intake diet caused a marked diuresis, low urine osmolality and low urinary urea concentration. It is assumed that the increase in interstitial substance between the vasa recta of the vascular bundle lowers the efficiency of the counter current barrier system for urea in the kidney and, as a consequence, the medullary urea gradient and urine concentrating capacity decreases. In animals on a high protein diet, the closely juxtaposed vasa recta assure an efficient countercurrent exchange, leading to accumulation and maintenance of a large urea gradient in the medulla and maximal urine concentration. It is suggested that the amount of interstitial material between the vasa recta of the vascular bundle might serve as a modulating factor for the urea concentration in the kidney.     

Location of an inducible nitric oxide synthase mRNA in the normal kidney

An inducible nitric oxide synthase (iNOS) mRNA was found primarily in the outer medulla of normal rat kidney. Identification of the mRNA was based upon the specificity of the oligonucleotide primers used for PCR amplification, PCR-Southern blot analysis and the nucleic acid sequence of the cloned PCR product. In addition to the outer medulla, glomeruli prepared from normal rat kidney contained significant amounts of an iNOS mRNA. These results suggest that there may be tonic influences in the outer medulla of the normal rat kidney resulting in the "steady-state" presence of an iNOS mRNA. Cortical tubules and the inner medulla were found to contain detectable but lesser amounts of the iNOS mRNA. The outer medulla was microdissected into proximal straight tubule (PST), medullary thick ascending limb (MTAL), medullary collecting duct (MCD) and vasa recta bundle (VRB). The iNOS mRNA was found primarily in the MTAL with minor amounts in the MCD and VRB of normal rat kidney. Animals were injected with lipopolysaccharide (LPS) and sacrificed 24 hours later. Treatment with LPS caused at least a 20-fold increase in the amount of iNOS mRNA in the liver or in macrophages isolated from the peritoneum. Endotoxin treatment led to over a 10-fold increase in iNOS mRNA content in glomeruli and the inner medulla. The iNOS mRNA level of the outer medulla was increased two- to threefold due to LPS treatment.     

Smaller sized particles are preferentially taken up by alveolar type II pneumocytes

Specific urea transporters are responsible for the rapid urea movements occurring in precise medullary structures of the mammalian kidney. Three of them, ensuring facilitated passive transports, have been cloned yet: UT2-long is responsible for the high vasopressin-dependent urea permeability of the terminal inner medullary collecting ducts; UT2-short is located along a short portion of the thin descending limbs of Henle's loops; UT11 is expressed along the descending vasa recta. These three transporters are involved in the accumulation of urea in the medulla, participating to the corticopapillary osmotic gradient required for urine concentration in the presence of antidiuretic hormone. UT2-long enables diffusion of urea in the inner medulla, and UT2-short and UT11 enable the recycling of this urea by counter-exchange. These transporters could also be involved in nitrogen balance by modulation of their expression according to the need for urea excretion (protein-rich diet), or for nitrogen conservation (protein-poor diet). Several other urea transporters, including active transporters responsible for urea secretion or reabsorption, remain to be cloned and characterized.     

Renal tubular and vascular urea transporters: influence of antidiuretic hormone on messenger RNA expression in Brattleboro rats

In the kidney, facilitated urea transport in precise vascular and tubular structures is mainly involved in water conservation. Three urea transporters have been cloned: UT2-long expressed in terminal inner medullary collecting duct (IMCD), UT2-short expressed in thin descending limb, and UT11 in descending vasa recta. The effect of arginine vasopressin (AVP) administration on mRNA expression of these three transporters was examined in Brattleboro rats with diabetes insipidus. V2 effects were discriminated from combined V1 + V2 effects by comparing treatments with 1-deamino-8-D-AVP (dDAVP) (selective V2 agonism) and AVP (V1 and V2 agonism). Acute and chronic treatments were studied. Abundance of specific mRNA was assessed by quantitative Northern blot analysis of RNA extracted from two regions of inner stripe of outer medulla and from two regions of inner medulla (IM). The results show that mRNA of these urea transporters are differently regulated by AVP. (1) Long-term treatment with either AVP or dDAVP does not alter UT2-long mRNA in tip IM (terminal IMCD) except for a transient initial decrease. (2) Unlike AVP, dDAVP induces the appearance of significant expression of UT2-long mRNA in base IM (initial IMCD), indicating a major V2 effect. (3) UT2-short mRNA in deep inner stripe of outer medulla and base IM (thin descending limb of short and long loops, respectively) is progressively upregulated with duration of AVP or dDAVP treatment. (4) The much higher changes in UT2-long and UT2-short induced by dDAVP compared with AVP suggest that they are dependent mainly on V2 agonism, and likely attenuated by V1 agonism. (5) UT11 mRNA expression in tip IM is equally depressed by AVP and dDAVP, indicating that this vascular transporter is also influenced by AVP and/or urine-concentrating activity, via an indirect mechanism that remains to be determined.     

Autoradiographic localization of vasopressin V1a receptors in the rat kidney using [3H]-SR 49059

Localization and characterization of binding sites of the selective non-peptide vasopressin receptor V1a ligand, [3H]-SR 49059, were investigated in the adult rat kidney by quantitative autoradiography using a fast-detecting radioluminographic phosphor-imaging plate system. [3H]-SR 49059, like the other V1a ligands used, showed a total absence of binding in the papilla, discrete and sparse labeling in the cortex and maximal binding in the outer part of the inner medulla. This labeling seemed to be mainly associated with medullary interstitial cells and vascular elements of the vasa recta. Conversely, [3H]-AVP intensely labeled the V2-enriched medulla-papillary portion of the kidney and, to a lesser extent, the cortical structures. [3H]-SR 49059 binding, quantified in the outer part of the inner medulla in rat kidney sections, was time-dependent, reversible, saturable and a single class of high affinity binding sites (Kd = 1.48 +/- 0.16 nM) was identified. The relative potencies of the reference peptide and non-peptide compounds to inhibit [3H]-SR 49059 binding confirm the V1a nature of the site and the stereospecificity of this binding. Thus, [3H]-SR 49059 allows the mapping and characterization of the V1a receptor population present in the rat kidney. The stability and the highly selective affinity of this non-peptide ligand for rat and human V1a receptors make it a suitable probe for the localization of V1a receptors in organs expressing heterogeneous populations of receptors.     

Localization and regulation of renal Na+/myo-inositol cotransporter in diabetic rats

We have examined the effect of diabetes on sodium/myo-inositol cotransporter (SMIT) mRNA levels and myo-inositol content in the kidney to test the hypothesis that diabetes-induced changes in renal myo-inositol levels are due to the regulation of SMIT mRNA levels. In streptozotocin-induced diabetic rats, after 3, 7 and 28 days of diabetes, SMIT mRNA levels in the whole kidney were increased three to fivefold, and remained increased by about twofold after six months of diabetes. Insulin treatment of diabetic rats normalized blood glucose levels and prevented the increase in SMIT mRNA levels. Treating diabetic rats with sorbinil, an aldose reductase inhibitor, corrected the abnormal accumulation of sorbitol but had no effect on the diabetes-induced increase in renal SMIT mRNA levels. The regional distribution of SMIT mRNA from normal rats showed a relative abundance in cortex, outer medulla, and inner medulla of 1.0:3.4:7.0. After seven days of diabetes, the levels of SMIT mRNA and myo-inositol content were significantly increased only in the outer medulla. In situ hybridization studies revealed that SMIT mRNA in the outer medulla was predominately localized to the medullary thick ascending limbs of Henle's loop and was not localized to any specific cell in the inner medulla. This distribution pattern was unchanged in diabetic rats. These studies show that diabetes causes an increase in renal SMIT mRNA, which is primarily localized to the outer medulla. Accumulation of myo-inositol by the thick ascending limb of Henle's loop may account for most of the increase caused by diabetes.     

Relationship between intrarenal distribution of blood flow and renin secretion during ureteral occlusion

The present study was undertaken to evaluate the relationship between renin secretion from the denervated kidney and intrarenal distribution of blood flow during reductions in renal perfusion pressure by partial constriction of the aorta with and without ureteral occlusion in the anesthetized dog. In addition, renin contents in different zones of the kidney were measured. A reduction in renal arterial pressure from normal pressure (125-135 mmHg) to 77 mm Hg resulted in significant increase in renin secretion and redistribution of cortical blood flow. A further reduction of renal arterial pressure to 51 mmHg produced a marked increase in renin secretion rate (RSR) without further changes in the intrarenal distribution pattern of blood flow. The pressure reductions during ureteral occlusion increased RSR without any change in the distribution pattern of blood flow, and a decrease in the amounts of extractable renin was found in the outer cortex of the experimental kidney. These findings suggest that renin release occurs mainly in the outer cortex, and this process may be stimulated when the mechanism of autoregulation fails as the perfusion presure approaches to the lower range of autoregulation in the outer cortex.     

Renal and intrarenal blood flow distribution in swine during severe exercise

It is known that a compensatory reduction and diversion of renal flow occurs in severe exercise in humans but not in dogs. We investigated this in miniature swine by measuring changes in total renal blood flow (TRF) and intra-renal blood flow (IRBF) distribution with tracer microspheres (15 +/- 5 mum) at rest and during steady-state exercise at 4.8-7.2 kph and 0% grade, and during severe exercise at 4.8-7.2 kph and 10% grade. We measured heart rate and cardiac output (Q) via implanted probes. TRF was determined as a percent of Q and as ml/100 g per min. IRBF was determined for the outer cortex, inner cortex, outer medulla, and inner medulla. Our results show that renal blood flow is significantly (P less than 0.05) reduced in pigs with exercise. Steady-state exercise reduced flow to about 66% of control and severe exercise reduced renal flow to 30% of control. IRBF was unchanged throughout. These results show that the exercising pig augments blood flow to skeletal muscle by reducing blood flow to kidneys, a response known to occur in man.     

Effect of oxygen and solute on PGE and PGF production by rat kidney slices

Increasing oxygen from 5% to 95% resulted in an increases production of both PGE's and PGF's. The release of prostaglandins from slices of rat kidney cortex and outer and inner medulla was measured. Prostaglandin production was observed predominantly in the inner medulla, was close to the lower limit of detection in the outer medulla, and was undetectable in the cortex. Increasing oxygen concentration resulted in a threefold increase in inner medullary prostaglandin production. Synthesis at 95% O2 was less at 2100 mOsm than at 300 mOsm, while synthesis at 5% O2 was not affected by high solute concentration. The implications of these results with respect to kidney function are discussed.     

The renal medulla and mechanisms of hypertension in the spontaneously hypertensive rat

A significant number of offspring from brother-sister matings of NIH-Okamoto-Aoki spontaneously hypertensive rats (SHRs) were found to be normotensive at 20 weeks of age. Over 20% of the animals that were hypertensive at this age had mild-to-moderate unilateral hydronephrosis at the time of sacrifice. In over 90% of the rats that did not develop hypertension spontaneously, ligation of one ureter raised blood pressure above 150 mm Hg within 2 weeks. In those rats made hypertensive by obstructing one ureter and in those that developed hypertension with accompanying naturally occurring hydronephrosis, subcutaneous implants of fragmented renal medulla from unrelated normal rats decreased blood pressure to normotensive levels. In contrast, medullary implants had no significant effect in rats developing hypertension spontaneously without hydronephrosis. Renal inner medullary plasma flow was low in the obstructed kidneys of hydronephrotic hypertensive SHRs but was elevated in the kidneys of nonhydronephrotic hypertensive SHRs. The hypertension in hydronephrotic SHRs appears to be related to an impairment of the antihypertensive function of the renal medulla. Such an impairment of medullary antihypertensive function does not appear to play a significant role in the hypertension in SHRs without hydronephrosis.     

Glucagon increases medullary interstitial electrolyte concentration in rat kidney

In anesthetized rats, tissue electrical admittance of the inner medulla (a measure of total ion concentration in the interstitium), medullary blood flow (laser Doppler technique), and renal clearances were measured simultaneously before and during i.v. infusion of glucagon at 110 and 330 ng.min-1.kg-1 body weight. Admittance increased modestly, 5.4% after a large glucagon dose (p < 0.01), whereas medullary blood flow was stable. Glomerular filtration rate increased transiently and then fell during high-dose glucagon infusion. The increase in tissue electrolyte (mostly NaCl) concentration in the medulla observed with stable medullary blood flow and decreasing glomerular filtration rate indicates that stimulation of NaCl reabsorption in the medullary ascending limb of Henle's loop by glucagon was the mechanism underlying augmentation of medullary ionic hypertonicity. This suggests that glucagon can contribute to the urine concentration process.     

Results on a pilot study of cultures of human lacteal secretions and benign and malignant breast tumors

The inhibitory action of indomethacin administered as a single-dose injection (4mg/kg) was examined under general anaesthesia in dogs, moderate volume expansion having been induced with physiological saline infusion. At 20 to 30 min after the administration of indomethacin, excretion of Na and water showed a fall of the same extent, GFR remaining stable and the effective plasma flow (CPAH) declining. RBF estimated by the 86Rb method decreased from 411 +/- 96 ml/min/100 g to 292 +/- 53 ml/min/100 g (p less than 0.01). This fall was coupled with an intrarenal redistribution of blood flow. While the cortical fraction of renal blood flow increased from 79% to 83.9% (p less than 0.001), its outer medullary fraction decreased from 17% to 13.2% (p less than 0.001) and its inner medullary fraction from 4.0% to 2.8% (p less than 0.05). The renal, primarily the medullary, vasculature is assumed on these grounds to be under the influence of a continuous secretion of prostaglandins which thus seem to be involved in the physiological control of intrarenal distribution of blood flow and of sodium and water excretion.     

3D time-resolved contrast-enhanced MR DSA: advantages and tradeoffs

To test whether renal V1-receptors selectively influence blood flow in the renal medulla, we compared the effects of infusion of [Phe2,Ile3,Orn8]vasopressin (3-30 ng/kg/min) by the intravenous, renal arterial, and renal medullary interstitial routes in anesthetized rabbits. Intravenous [Phe2,Ile3,Orn8]vasopressin (30 ng/kg/min) reduced renal medullary perfusion (MBF) by 36 +/- 5% but did not significantly affect cortical perfusion (CBF). MBF was also reduced with the renal arterial (35 +/- 5%) and renal medullary interstitial (40 +/- 7%) routes but, in contrast to the intravenous infusion, CBF was also reduced, by 21 +/- 3% and 15 +/- 3%, respectively. Urine flow and sodium excretion were increased by [Phe2,Ile3,Orn8]vasopressin, and with direct intrarenal administration, this effect was similar for both the infused (left) and noninfused (right) kidneys. After a 20-min renal medullary interstitial infusion of [3H]norepinephrine, radiolabel concentration was approximately fivefold greater in the left medulla than in the left cortex. We conclude that [Phe2,Ile3,Orn8]vasopressin acts on V1-receptors to alter regional kidney blood flow and tubular salt and water handling. The V1-receptors involved are almost certainly within the kidney itself, but given the contrasting effects of the different infusion routes on MBF and CBF, we cannot exclude the possibility that some of the observed effects of [Phe2,Ile3,Orn8]vasopressin are mediated by activation of extra-renal V1-receptors.     

Cardiorespiratory adaptations induced by aerobic training in middle-aged men: the importance of a decrease in sympathetic stimulation for the contribution of dynamic exercise tachycardia

The influence of diuresis and antidiuresis on the expression of heat shock proteins (HSP) 25, 60, 72 and 73 in the renal cortex and outer and inner medulla of Wistar rats was analysed. Medullary osmolality was reduced by long-term diuresis (3% sucrose in the drinking water for 3 weeks) and subsequently enhanced by transition to a concentrating state by giving normal drinking water again in combination with deamino-D-arginine vasopressin (dDAVP) for 5 days. Western blot analyses revealed that neither HSP73 nor HSP60 was influenced by any treatment. The HSP72 level in the medulla was markedly reduced (50%) when osmolality was lowered and increased when tonicity was high. RNAse protection assays showed that the effects on HSP72 are parallelled in general by changes in HSP72 mRNA. While levels of HSP25 were not influenced, isoelectric focusing revealed that the degree of phosphorylation of outer and inner medullary HSP25 increased following both treatments. It thus seems that HSP73 and HSP60 are not directly involved in the long-term adaptation to varying medullary osmolalities. The correlation between changes in osmolality and amounts of the major stress-inducible HSP72 in the medulla implies that medullary hypertonicity is stressful for kidney cells. Furthermore, adaptation to pronounced changes in the osmolality of the environment most likely involves phosphorylation of HSP25.     

Two-chain factor VIIa generated in the pericardium during surgery with cardiopulmonary bypass : relationship to increased thrombin generation and heparin concentration

The effect of changes in medullary extracellular tonicity on mRNA expression for aldose reductase (AR), sorbitol dehydrogenase (SDH), Na+/Cl-/betaine (BGT) and Na+/myo-inositol (SMIT) cotransporter in different kidney zones was studied using Northern blot analysis and non-radioactive in situ hybridization in four groups of rats: controls, acute diuresis (the loop diuretic furosemide was administered), chronic diuresis (5 days of diuresis), and antidiuresis [5 days of diuresis followed by 24 h deamino-Cys1, d-Arg8 vasopressin (dDAVP)]. Acute administration of the loop diuretic furosemide significantly reduced AR, SMIT and BGT gene expression in the inner and outer medulla compared with controls. Administration of dDAVP to chronically diuretic rats raised the expression of these three mRNAs in the inner but not the outer medulla compared with the chronically diuretic rats. None of these alterations in medullary tonicity significantly changed SDH expression. The in situ hybridization studies showed AR, BGT and SMIT mRNAs to be expressed in both epithelial and non-epithelial cells of the outer and inner medulla. The various cell types (epithelial, endothelial and interstitial cells) differed in their expression pattern and intensity of AR, SDH, BGT and SMIT mRNA, but the inner medullary cells responded uniformly to a decrease in extracellular tonicity with a reduction, and to an increase with enhancement of their AR, BGT and SMIT expression.     

Expression of aquaporin-1 (AQP1) in the adult and developing sheep kidney

The distributions of aquaporin-1 mRNA and protein were studied by hybridization histochemistry with a homologous riboprobe and immunohistochemistry, in the adult sheep kidney. Heaviest labelling occurred in the thin descending limb (DTL) of the loop of Henle in the inner stripe of the outer medulla, with apparent decreasing expression in the inner medulla, outer stripe of the outer medulla and cortex, but no quantitation was performed. Only proximal tubules (PT) (convoluted and straight) and DTL labelled. The glomerulus showed no labelling, consistent with the pattern in the rat but different to that in the human. During ontogeny, no labelling occurred in the mesonephros at 27 or 41 days of gestation (term = 145-150 days) but other structures did label at 27 days (heart, lung bud, blood vessels surrounding developing spinal cord). Labelling first occurred faintly in the metanephros at 41 days of gestation and increased throughout gestation consistent with morphological development of nephrons.