Mammalian-type Nephrons Research Articles

3D morphometric study of domestic fowl glomerulus-A scanning electron microscope study of vascular corrosion casts.

Microvascularization of domestic fowl kidneys was studied using scanning electron microscopy (SEM) of vascular corrosion casts (VCCs). Two types of nephrons, mammalian-type (MT) and reptilian-type (RT) nephrons and their glomerular structure were analysed quantitatively by 3D morphometry. A significant difference in shape and size between the MT and RT glomeruli was found. The mean diameter of the RT glomeruli was about 56µm, while that of MT glomeruli was significantly larger, namely about 80µm. The afferent arterioles in mammalian-type glomeruli usually bifurcated into two lobular branches and formed a complex glomerular capillary network with numerous loops. Reptilian-type glomeruli consisted of a single capillary forming few loops and leaving the glomerulus as efferent arteriole. Diameters of afferent and efferent arteriolar replicas were similar in all three kidney divisions of MT and RT nephrons. The absence of the interconnecting branches between the MT nephron capillaries at the gross inspection suggests that the mammalian-type nephron glomeruli, although more complex than the reptilian type, are not equivalent to those in mammalian kidneys.

Basolateral regulation of pHi in proximal tubules of avian loopless and long-looped nephrons in bicarbonate

In isolated, nonperfused chicken proximal tubules from both loopless reptilian-type and long-looped mammalian-type nephrons, resting intracellular pH (pHi), measured with pH-sensitive fluorescent dye 2',7'-bis(2-carboxyethyl)-5,6-carboxyfluorescein (BCECF), was approximately 7.1 under control HCO3- conditions [20 mM N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES)/5 mM HCO3(-)-buffered medium with pH 7.4 at 37 degrees C] and was reduced to approximately 6.8 in response to NH4Cl pulse. The rate of recovery of pHi (dpHi/dt) from this level to the resting level in proximal tubules from both nephron types was (1) significantly reduced by the removal of Na+ or both Na+ and Cl- from the bath, and (2) unaffected by the removal of Cl- from the bath or the presence of a high K+ concentration or Ba2+ in the bath. In proximal tubules from long-looped mammalian-type, but not loopless reptilian-type, nephrons, dpHi/dt was significantly reduced by the addition of either 5-(N-ethyl-N-isopropyl) amiloride (EIPA) or 4,4'-diisothiocyanostilbene-2,2'disulfonate (DIDS) to the bath. These data suggest that a Na+/H+ exchanger and most likely a Na(+)-dependent Cl-/HCO3- exchanger are involved in basolateral regulation of pHi in mammalian-type nephrons whereas none of the commonly identified basolateral acid-base transporters appear to be involved in regulation of pHi in reptilian-type nephrons.

Basolateral regulation of pHi in proximal tubules of avian loopless and long-looped nephrons in bicarbonate.

In isolated, nonperfused chicken proximal tubules from both loopless reptilian-type and long-looped mammalian-type nephrons, resting intracellular pH (pHi), measured with pH-sensitive fluorescent dye 2',7'-bis(2-carboxyethyl)-5,6-carboxyfluorescein (BCECF), was approximately 7.1 under control HCO3- conditions [20 mM N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES)/5 mM HCO3(-)-buffered medium with pH 7.4 at 37 degrees C] and was reduced to approximately 6.8 in response to NH4Cl pulse. The rate of recovery of pHi (dpHi/dt) from this level to the resting level in proximal tubules from both nephron types was (1) significantly reduced by the removal of Na+ or both Na+ and Cl- from the bath, and (2) unaffected by the removal of Cl- from the bath or the presence of a high K+ concentration or Ba2+ in the bath. In proximal tubules from long-looped mammalian-type, but not loopless reptilian-type, nephrons, dpHi/dt was significantly reduced by the addition of either 5-(N-ethyl-N-isopropyl) amiloride (EIPA) or 4,4'-diisothiocyanostilbene-2,2'disulfonate (DIDS) to the bath. These data suggest that a Na+/H+ exchanger and most likely a Na(+)-dependent Cl-/HCO3- exchanger are involved in basolateral regulation of pHi in mammalian-type nephrons whereas none of the commonly identified basolateral acid-base transporters appear to be involved in regulation of pHi in reptilian-type nephrons.

Functional Receptors in the Avian Kidney for C-Type Natriuretic Peptide

Renal actions of avian-specific C-type natriuretic peptide (chCNP) were investigated in the conscious Pekin duck. Under conditions of steady-state renal water and salt elimination, systemic chCNP administration (6 and 30 pmol/min·kg BW for 20 min) dose dependently induced transient natriuresis and diuresis. Mean arterial pressure and heart rate remained constant throughout the experiment. Employing receptor autoradiography, binding sites specific for[ 125I]BH-chCNP could be localized at high density in glomeruli of both reptilian- and mammalian-type nephrons, and arterioles of the avian kidney. The distal tubular zone revealed[ 125I]BH-chCNP binding sites at medium, the medullary cone area at low density. Using an enriched kidney membrane fraction, competitive displacement studies with [125I]BH-chCNP as radioligand and various unlabeled peptide analogs (chANP, chCNP, rANP, rBNP, frANP, rANP(4–23)) allowed the discrimination of high-affinity (IC50 values 10−10–10−9m) and low-affinity (IC50 values 10−8–10−7m) binding sites different from typical mammalian receptor subtypes. Intracellular cyclic GMP formation could be demonstrated immunocytochemically for both types of glomeruli and cells of the distal tubular zone in fixed tissue sections after in vivo application of chCNP (0.8 nmol/min·kg BW; 5 min). The results obtained by combination of physiological in vivo studies and in vitro receptor analysis indicate an important role for chCNP in the modulation of avian kidney function.

Functional receptors in the avian kidney for C-type natriuretic peptide.

Renal actions of avian-specific C-type natriuretic peptide (chCNP) were investigated in the conscious Pekin duck. Under conditions of steady-state renal water and salt elimination, systemic chCNP administration (6 and 30 pmol/min x kg BW for 20 min) dose dependently induced transient natriuresis and diuresis. Mean arterial pressure and heart rate remained constant throughout the experiment. Employing receptor autoradiography, binding sites specific for [125I]BH-chCNP could be localized at high density in glomeruli of both reptilian- and mammalian-type nephrons, and arterioles of the avian kidney. The distal tubular zone revealed [125I]BH-chCNP binding sites at medium, the medullary cone area at low density. Using an enriched kidney membrane fraction, competitive displacement studies with [125I]BH-chCNP as radioligand and various unlabeled peptide analogs (chANP, chCNP, rANP, rBNP, frANP, rANP(4-23)) allowed the discrimination of high-affinity (IC50 values 10(-10)-10(-9) M) and low-affinity (IC50 values 10(-8)-10(-7) M) binding sites different from typical mammalian receptor subtypes. Intracellular cyclic GMP formation could be demonstrated immunocytochemically for both types of glomeruli and cells of the distal tubular zone in fixed tissue sections after in vivo application of chCNP (0.8 nmol/min x kg BW; 5 min). The results obtained by combination of physiological in vivo studies and in vitro receptor analysis indicate an important role for chCNP in the modulation of avian kidney function.

Regulation of intracellular pH in proximal tubules of avian long-looped mammalian-type nephrons.

In nonperfused proximal tubules isolated from chicken long-looped mammalian-type nephrons, intracellular pH (pHi), measured with the pH-sensitive fluorescent dye 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein, was approximately 7.3 under control conditions (HEPES-buffered medium with pH 7.4 at 37 degrees C) and was reduced to approximately 7.0 in response to NH4Cl pulse. The rate of recovery of pHi from this level to the resting level was 1) significantly reduced by the removal of Na+ from the bath, 2) significantly increased by the removal of Cl- from the bath, 3) unchanged by the removal of both Na+ and Cl- from the bath, 4) significantly reduced by the addition of either ethylisopropylamiloride or DIDS to the bath, 5) significantly increased by a high bath K+ concentration, and 6) unchanged by the addition of Ba2+ to the bath. These data suggest that both Na+-coupled and Cl--coupled basolateral acid-base fluxes are involved in determining the rate of recovery of pHi after acidification. The most likely ones to be important in regulating pHi are a Na+/H+ exchanger and a Na+-coupled Cl-/HCO-3 exchanger. In birds, long-looped mammalian-type nephrons resemble short-looped transitional nephrons but differ markedly from superficial loopless reptilian-type nephrons.

Control of sodium and chloride transport in the thick ascending limb in the avian nephron

We previously reported that birds may concentrate urine by a countercurrent multiplier mechanism using single-solute (NaCl) recirculation, in which the thick ascending limb of Henle (TAL) of mammalian-type nephrons provides an energy source. The Japanese quail TAL has a higher lumen-to-bath Cl flux (JCl,lb) than that of mammals; the mechanism for maintaining the osmotic gradient along the medullary cone is unknown. We investigated whether salt delivery alters the NaCl reabsorption rate in the TAL dissected from both normal and salt-loaded Japanese quail, Coturnix coturnix, 3-7 wk old. When salt loading to the TAL was increased by increasing the perfusion flow rate (PFR), the lumen-to-bath Na and Cl flux coefficients (KNa,lb or KCl,lb, 10(-7) cm2/s) increased, respectively (P < 0.05), from 5.1 +/- 0.9 to 7.6 +/- 0.7 and from 6.8 +/- 0.9 to 8.9 +/- 1.4. Ouabain addition significantly reduced the KNa,lb. A significant correlation existed between PFR and JCl,lb (r = 0.69, P < 0.01) and PFR and JNa,lb. When the TAL was perfused at a low PFR, increasing Cl concentration in the perfusate and bathing medium from 125 to 175 (P < 0.05) or 225 mM (P < 0.01) increased JCl,lb but not KCl,lb, while decreasing Cl concentration decreased JCl,lb but not KCl,lb. Salt loading of intact birds (0.2 M NaCl drinking water) for 7 days increased the plasma Na level and the cloacal fluid-to-plasma osmolality ratio from 0.28 +/- 0.07 to 1.06 +/- 0.05 (P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Micropuncture study of avian kidney: effect of prolactin.

Renal function was studied in anesthetized European starlings (Sturnis vulgaris) by micropuncture and clearance techniques. A previous study had shown some changes in the renal function of birds in molt. Endogenous prolactin levels have been shown to be elevated at this time. In the present study, renal function was studied during the infusion of an isotonic sodium chloride solution and then when ovine prolactin was added to the infusion. Ovine prolactin resulted in an elevation of plasma urate concentration but had no significant effects on other plasma parameters or on the concentration of the various ions in tubular fluid samples. Urine flow rate was significantly higher during prolactin infusion, although whole animal glomerular filtration rate, single nephron glomerular filtration rate, and the percentage fluid reabsorption in the proximal tubule (up to the point of micropuncture) were unaltered. The whole animal fractional excretions of sodium and chloride were significantly higher, those of magnesium and potassium were significantly lower, and the fractional excretion of phosphate tended to be lower during prolactin infusion. At the level of the proximal tubules of the reptilian-type nephrons, prolactin infusion caused a slight reduction in the net reabsorption of sodium and chloride. Therefore the main effects of prolactin on renal function are occurring in the distal nephron of the reptilian-type nephrons or in the mammalian-type nephrons. It appears that prolactin may affect renal function in avian species and that these actions may be modulated by a range of factors as has been demonstrated in mammals.

Cellular basis of an avian countercurrent multiplier system

A kidney from the budgerigar (budgie, parakeet; Melopsittacus undulatus) is composed of cortical reptilian-type nephrons (without loops of Henle) and mammalian-type nephrons (with loops) grouped together in medullary cones. The loop of the mammalian-type nephrons has a descending segment composed of thin and highly interdigitated cells. These thin limb cells have few mitochondria (15% of cell volume), undetectable Na+,K(+)-ATPase activity, and virtually no basolateral surface amplification. Prior to the hairpin turn, the descending limb thickens, but the cells continue to lack basolateral amplification. Cells just prior to and within the hairpin turn resemble cells of the entire ascending limb. These cells are thick (there is no thin ascending segment in the avian loop), with extensive infoldings of the basolateral membrane surrounding numerous mitochondria (45% of cell volume). The area of basolateral membrane is 25 times that of the apical membrane. The basolateral membrane (but not the apical membrane) is enriched in Na+,K(+)-ATPase activity. The structure of the avian mammalian-type nephron (as epitomized by the budgie nephron) and the fact that NaCl accounts for over 90% of the osmotic activity of avian urine leads to the conclusion that the countercurrent multiplier of the avian kidney functions by active NaCl transport from the entire ascending limb. No explanation is offered for the transport specializations found in the thick descending segment of the loop, just prior to the hairpin turn.

Sodium chloride and water transport in the thin descending limb of Henle of the quail.

Birds and mammals can produce hyperosmotic urine, but their renal morphology and urine-concentrating mechanisms differ. To elucidate the countercurrent urine concentration mechanism in birds, we examined the structure and transport properties of the descending limb (DL) of Henle of mammalian-type nephrons in Japanese quail, Coturnix coturnix. In the avian renal medulla, a prominent ring of collecting ducts and scattered thick limbs surrounds a core of capillaries and DLs. Epithelial cells in the upper DL (DLu) have abundant microvilli and shallow, tight junctions; cells in the lower DL are flat and have little interdigitation. Transepithelial voltage was zero when the DLu was perfused and bathed in isosmotic avian Ringer solution. The efflux coefficients (10(-7) cm2/s) for Na (31.7 +/- 2.3) and Cl (24.9 +/- 3.6) were not significantly different and were unaltered by ouabain (10(-4) M) (32.5 +/- 2.2). Diffusional water permeability measured by [3H]H2O was low (73.0 +/- 7.8, 10(-7) cm2/s). Volume flux was nearly zero and increased only slightly when an osmotic gradient was imposed. These results suggest the DLu is highly permeable to Na and Cl and virtually impermeable to water; thus NaCl extruded actively from the thick ascending limb may enter the DL unaccompanied by water. This countercurrent multiplication system by use of single-solute recycling and a transport cascade of graded hairpin turns may help establish an osmotic gradient along the medullary cone. Thus avian and mammalian renal countercurrent multiplication systems may differ.

Post-hatching growth of the kidney in the chukar (Aves: Phasianidae).

The post-hatching growth of the kidney of the chukar partridge, Alectoris chukar, was studied in birds aged 5, 24, 56, and 140 days. As body mass increased from 24 gm (5 d) to 479 gm (140 d), kidney mass increased from 0.16 to 1.6 gm. The number of nephrons in the kidney increased through age 56 d, but did not change thereafter. This increase was primarily a function of increasing development of the cortical regions; more than 90% of reptilian-type nephrons developed after hatching. The full complement of medullary cones was present at hatching. However, the lengths of the medullary cones and the number of loops of Henle of mammalian-type nephrons contained in each medullary cone increased through growth, so that approximately 65% of the mammalian-type nephrons developed after hatching. The chukar kidney contained 19% mammalian-type nephrons at age 5 d, but this was reduced to approximately 5% by 24 d, after which the proportions of mammalian-and reptilian-type nephrons did not change. This study demonstrates that metanephric kidney growth in birds begins in the deepest regions of the kidney and then extends peripherally but also that mammalian-type nephrons continue to develop throughout kidney growth, presumably from pre-existing reptilian-type nephrons.

Diluting segment in avian kidney. II. Water and chloride transport.

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.

Proportions of mammalian-type and reptilian-type nephrons in the kidneys of two passerine birds.

We compared the proportions of mammalian-type and reptilian-type nephrons in the kidneys of two species of passerine birds. The desert house sparrow (Passer domesticus) is relatively well adapted for water conservation, whereas the white-crowned sparrow (Zonotrichia leucophrys) is more mesic adapted. The two species do not differ in body mass, but the kidneys of P. domesticus are significantly smaller than those of Z. leucophrys. Associated with its smaller size, the house sparrow kidney has significantly fewer glomeruli (35,700 per kidney) than does the white-crowned sparrow kidney (53,000 per kidney). The medullary cones, which contain the loops of Henle of the mammalian-type nephrons, are significantly longer in house sparrows than in white-crowned sparrows (2.2 vs. 1.9 mm). The number of medullary cones, the number of nephrons per medullary cone, and, hence, the number of mammalian-type nephrons do not differ between the two species. The smaller number of nephrons in the kidney of the house sparrow therefore represents a smaller number of reptilian-type nephrons. Desert house sparrows have 18% mammalian-type nephrons, whereas white-crowned sparrows have 10% mammaliantype nephrons. The relative reduction of reptilian-type nephrons in P. domesticus may reduce the flow of dilute urine through the collecting ducts, thereby permitting a greater concentration gradient to be established along the length of the medullary cones.

Endocrine regulation of avian renal function.

The avian kidney contains reptilian-type (RT) nephrons that do not function in concert and mammalian-type (MT) nephrons that do function in concert to permit production of concentrated urine. Significant data on endocrine regulation of renal function are available only for antidiuretic hormone (arginine vasotocin, AVT) and parathyroid hormone (PTH). Plasma AVT increases and number of filtering RT nephrons decrease when plasma osmolality increases. Small doses of AVT reduce the number of filtering RT nephrons, and all RT nephrons filter following acute neurohypophysectomy. In addition to changing tubule permeability to water, AVT affects the concentrating mechanism by reducing the number of filtering RT nephrons and flow through collecting ducts. Both net tubular reabsorption and net tubular secretion of PO4 occur. Parathyroidectomy (PTX) stimulates PO4 reabsorption, and PTH replacement decreases reabsorption and stimulates secretion, apparently in proximal tubules of RT nephrons. PTX stimulates and PTH replacement inhibits Na-dependent PO4 transport by renal brush border membrane vesicles. PTH also stimulates Ca2+ reabsorption but apparently not long the proximal tubule of RT nephrons.

Renal handling of sodium chloride and its control in birds.

Avian kidneys show features of both mammalian and non-mammalian kidneys, possessing cortical reptilian-type (RT) and medullary mammalian-type (MT) nephrons, blood supplies from renal arterial and renal portal systems, and a primitive macula densa. To determine how these morphological characteristics contribute to unique renal functions, we examined first a possible functional link between the renal tubules and the preglomerular vasculature and, second, the function of the loop of Henle of the MT nephrons. Infusion of 5% NaCl (2.0 ml/kg/h) into the renal portal system of the pullet Gallus domesticus caused diuresis and natriuresis in the infused side, whereas infusion of the same dose into the systemic circulation showed no effect. Infusion of 10% NaCl at a higher flow rate into either the renal portal or systemic routes caused hypernaturemia and a prolonged antidiuresis, presumably due to a release of arginine vasotocin. Plasma renin activity (PRA) decreased during renal portal infusion of hypertonic saline, suggesting that increases in intratubular or peritubular NaCl levels may suppress PRA. The thick limb of the loop of Henle (TLH) isolated from the MT nephron of the quail Coturnix coturnix revealed a lumen-positive, furosemide-sensitive transepithelial voltage (Vt) that requires the presence of both Na and Cl. Net water flux (Jv) was nearly zero when the TLH was perfused and bathed with isosmotic solution. Net water flux increased only slightly and osmotic water permeability (Lp) was low when the osmotic gradient was imposed. Vasotocin altered neither Vt, Jv, nor Lp. Chloride efflux was higher than Cl influx, and net Cl absorption was comparable to that of the TLH of the mammalian kidney.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of water load on renal glomerular and tubular function in desert quail.

Effects of an acute intravenous water load on glomerular and tubular function were studied in the anesthetized desert quail Lophortyx gambelii. Total-kidney glomerular filtration rate (GFR) and single-nephron glomerular filtration rates (SNGFR) of reptilian-type and mammalian-type nephrons increased by more than 50% compared with the GFR and SNGFRs measured during a control mannitol diuresis. Despite the increase in GFR, urine flow rate was only 56% of that in control mannitol diuresis, free-water clearance (CH2o) remained at 1-2% of GFR, and the animals excreted only about 79% of the water load. More than 99% of the filtered sodium and 93% of the filtered water were reabsorbed during the water load. Possible reasons for the increased GFR and the failure to produce a dilute urine or excrete the water load are discussed.

Relative thickness of the renal medulla in birds.

The intraspecific mean length of medullary cones in avian kidneys is analogous to medullary thickness in the mammalian kidney. Hence, relative medullary thickness (based upon kidney volume) can be calculated for birds as was done in mammals years ago. Comparative figures are given for 26 species from nine avian orders. The organizational pattern of cortex and medulla in the bird kidney is reviewed, and a simplified diagram of this relationship is presented. With some exceptions, urine concentrating ability and relative medullary thickness are directly proportional in mammals. Contrarily, no similar trend was evident in birds when current information on water economy was compared to relative medullary thickness in various species. There are a number of factors (such as the respective functional roles of reptilian and mammalian-type nephrons, interspecific variations in ion transport, etc.) which require study before the significance of relative thickness in the avian medulla can be evaluated more thoroughly.