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T L Pallone

Publications and source records attributed to T L Pallone.

At least 37 records · Page 2Linked to original sources
PubMed1996

Adenosine modulates vasomotor tone in outer medullary descending vasa recta of the rat.

Adenosine is generated within the renal medulla under hypoxic conditions and is known to induce net vasoconstriction within the renal cortex while increasing medullary blood flow and oxygenation. To test the hypothesis that vasoconstriction of outer medullary descending vasa recta (OMDVR) is modulated by adenosine, we examined the effects of adenosine and adenosine Al and A2 receptor subtype agonists on in vitro perfused control and preconstricted rat OMDVR. Constriction with angiotensin II (ANG II, 10(-9) M) was attenuated by adenosine in a concentration-dependent manner (EC50 = 2.0 x 10(-7)M, P < 0.05). Similarly, an adenosine A2 agonist (CGS-21680, 10(-7) M), but not an adenosine Al agonist (cyclohexyladenosine, 10(-6) M), attenuated ANG II-induced vasoconstriction. Under control conditions, ablumenal application of adenosine (10(-12) to 10(-5) M) elicited a biphasic response. Additionally, cyclohexyladenosine (10(-6) M) caused vasoconstriction and CGS-21680 (10(-6) M) had no effect on untreated vessels. Finally, an influence of ANG II receptor stimulation on adenosine Al receptor-mediated vasoconstriction could not be shown. These data suggest that OMDVR possess both Al and A2 adenosine receptors and that they mediate constriction and dilatation, respectively. We conclude that adenosine is a potent modulator of OMDVR vasomotor tone and that its net effect is dependent upon local concentrations.

Adenosine↗
PubMed1995

Effect of norepinephrine and acetylcholine on outer medullary descending vasa recta.

To examine their responsiveness to norepinephrine (NE) and acetylcholine (ACh), outer medullary descending vasa recta (OMDVR) have been dissected from vascular bundles of the rat and perfused in vitro. Abluminal application of NE produced graded vasoconstriction in a concentration range of 10(-9)-10(-6) M. When applied with NE, ACh at concentrations of 10(-8)-10(-5) M dilated NE-preconstricted OMDVR. In contrast, ACh applied in the absence of NE caused vasoconstriction. ACh-induced vasodilation was blocked by addition of the nitric oxide synthase inhibitor N omega-nitro-L-arginine (L-NNA, 2 x 10(-4) M). L-NNA in the absence of ACh enhanced NE-induced vasoconstriction. Supraphysiological (10(-3) M) L-arginine (L-Arg) reversed the effects of L-NNA, and abluminal application of L-NNA alone resulted in OMDVR vasoconstriction. At concentrations of 10(-6)-10(-3) M, abluminal application of L-Arg produced graded vasodilation of NE-constricted OMDVR. These results suggest that adrenergic and cholinergic innervation could influence OMDVR vasomotor tone to modulate total and regional blood flow to the renal medulla. The data also favor a role for the activity of constitutively expressed nitric oxide synthase to modulate OMDVR vasoactivity.

Acetylcholine↗
PubMed1995

Diffusive transport of solute in the rat medullary microcirculation.

Outer medullary descending vasa recta (OMDVR) permeability to sodium (PNa) is much lower than to urea (Purea). Based on these findings, we hypothesized that sodium and urea diffuse across the OMDFR wall by separate routes. To further test this, we simultaneously perfused OMDVR with 22Na and 36Cl, [14C]urea, [3H]raffinose, or tritiated water. The permeability of OMDVR to 22Na and [3H]raffinose was found to increase markedly and reversibly with perfusion rate. PNa was highly correlated with the permeability to Cl (PCl) and to [3H]raffinose (Praf) (R = 0.90 and 0.95, respectively) but not with Purea (R = 0.23). Praf was also correlated with inulin permeability (PIn) (R = 0.93). The intercepts for the regressions of PNa with PCl and Praf and for Praf with PIn were zero. In contrast, OMDVR with low PNa retained very high diffusional water permeability (PD) and Purea, a finding consistent with separate routes for permeation of those tracers. We previously established that thiourea is a competitive inhibitor of OMDVR urea transport. In the presence of 100 mM thiourea, OMDVR PNa and Purea were correlated (R = 0.71) but retained an intercept much > 0. We conclude that Na, Cl, raffinose, and inulin are likely to traverse the OMDVR wall through a common pathway, whereas specific mechanisms exist to regulate the permeation by urea and water.

Animals↗
PubMed1995

Prostaglandin E2 abrogates endothelin-induced vasoconstriction in renal outer medullary descending vasa recta of the rat.

Endothelins (ET) and prostaglandin E2 are synthesized in the inner medulla by collecting duct epithelium and interstitial cells, respectively. All ascending vasa recta (AVR) blood returns from the inner medulla to the cortex in outer medullary vascular bundles. We reasoned that hormones might influence medullary blood flow by diffusing across AVR fenestrations to modulate vasoconstriction of outer medullary descending vasa recta (OMDVR). To investigate this possibility, OMDVR dissected from vascular bundles were exposed to ET-1, 2, or 3. Each endothelin isoform induced stable vasoconstriction with potency, ET-1 > ET-2 > ET-3 (EC50, 1.8 x 10(-15), 5.9 x 10(-12), and 8.8 x 10(-10) M, respectively). The ETA receptor antagonist BQ-123 and BQ-610 (10(-6) M), as well as an ETA and ETB receptor antagonist combination, attenuated vasoconstriction due to ET-1 (10(-12) M). BQ-123 had no effect on the response to ET-3 (10(-8) M). The ETB receptor antagonist BQ-788 (10(-6) M) attenuated the response to ET-3 (10(-10) M), but not that to ET-1 (10(-12) M). Finally, PGE2 (10(-6) M) reversibly dilated OMDVR preconstricted with ET-1 (10(-12) M) or ET-3 (10(-8) M) but not ET-1 (10(-10) M). We conclude that ET-1,2, and 3 are potent constrictors of OMDVR and the response to ET-1 is mainly ETA receptor subtype mediated, while ET-3 acts via the ETB. PGE2 modulates ET induced constriction. These findings are consistent with interactive feedback and control of medullary perfusion by locally synthesized hormones.

Animals↗
PubMed1994

Extravascular protein in the renal medulla: analysis by two methods.

Two methods have been used to test for the presence of extravascular protein in the interstitium of the renal inner medulla. First, ascending vasa recta (AVR) segments were perfused with buffer containing 5 g/dl of albumin. The hydraulic pressure in the perfused vessel was varied to control transmembrane volume flux (Jv) to the interstitium. Interpolation to the point of zero Jv was employed to estimate interstitial Starling forces in the hydropenic rat papilla. Analysis of those experiments predicts that interstitial protein concentration (Ci) is high. When AVR segments are filled with oil, the oil column spontaneously breaks up as fluid is secreted into the lumen from the papillary interstitium. To obtain a lower limit on Ci, isolated AVR segments (IAS) filled with oil were sampled to measure protein concentration in the secreted fluid. In hydropenic rats, protein concentration was 3.4 +/- 0.5 and 5.2 +/- 0.2 g/dl in IAS and adjacent free-flowing AVR, respectively (P < 0.01). In rats subjected to furosemide and saline diuresis, the values were nearly identical, 4.7 +/- 0.2 and 5.2 +/- 0.2 g/dl, respectively. These separate experimental approaches corroborate a high concentration of protein in the renal inner medullary interstitium.

Animals↗
PubMed1994

Characterization of the urea transporter in outer medullary descending vasa recta.

Partially because of facilitated transport of urea, urea permeability (Pu) of the outer medullary descending vasa recta (OMDVR) frequently exceeds sodium permeability by more than an order of magnitude. This study characterizes the OMDVR urea transporter. Application of the urea analogue thiourea (200 mM) to the abluminal surface of microperfused OMDVR inhibited Pu by 33%. When osmolarity due to thiourea was balanced by addition of mannitol or thiourea, similar results were obtained. Thiourea produced graded inhibition of Pu from 343 +/- 54 (SE) to 191 +/- 43 x 10(-5) cm/s as concentration was increased from 0 to 100 mM. The thiourea concentration needed for half-maximal inhibition was 19 mM. The abilities of urea analogues to reduce Pu were compared by addition of 50 mM concentrations to the bath and perfusate. Thiourea and methylurea produced 32 and 34% inhibition of Pu, respectively, whereas urea and acetamide produced only 3 and 11% inhibition, respectively. The transporter showed negligible saturation as the transmural urea gradient was increased from 0 to 200 mM. Phloretin and p-chloromercuribenzenesulfonate inhibited Pu in a concentration-dependent fashion. It is concluded that a transporter confers high Pu to OMDVR. Pu is equally high when measured by urea influx or efflux. Properties of the transporter are similar to those expressed by the inner medullary collecting duct.

4-Chloromercuribenzenesulfonate↗
PubMed1994

Hyperosmolality and sperm storage in hibernating bats: prolongation of sperm life by dehydration.

Osmolalities of epididymal fluids obtained by micropuncture from hibernating species of bats (Myotis lucifugus) rise during sperm storage periods to as high as 1,523 mmol/kgH2O (approximately 5 times that of plasma). In vitro studies establish that hyperosmolality can preserve viability and prevent initiation of progressive motility in bat epididymal spermatozoa as well as induce their quiescence by reducing respiration. Reduction of osmolality (to 500-600 mmol/kgH2O) induces swelling of sperm and allows the initiation of motility and increased metabolic rate; further reduction of osmolality to < 300 mmol/kgH2O compromises permeability barriers and causes loss of motility. We hypothesize that seasonal establishment of hyperosmotic conditions driven by those cells that constitute the limits of the epididymal lumen dehydrates the compliant spermatozoa and thereby minimizes their metabolic needs. A novel form of cell storage dependent on unique adaptations of the epididymal epithelium for solute and water transport is implicated. To date, the operative osmolyte or osmolytes responsible for elevating osmolality in this system remain elusive.

Animals↗
PubMed1994

A simplified device for injection of paraffin wax blockades.

Microperfusion of renal tubules and microvessels in vivo requires occlusion of the lumen with a reliable blockade. Paraffin wax serves this purpose but requires specialized hydraulic or mechanical devices to stably hold a micropipette while this solid material is extruded through micron-sized tips. This communication describes the construction of a simple and very inexpensive wax injector from commercially available parts. The rear of an aluminum holder is threaded so that a thumbscrew can be used to push a stiff wire into the lumen of a micropipette. A solder cast of the micropipette lumen is employed as a tight-fitting piston to generate the high pressures required for wax extrusion. Several wax injectors can be made at negligible cost. This device has been used in this laboratory to introduce over 1,000 wax blocks into papillary vasa recta.

Animals↗
PubMed1994

Vasoconstriction of outer medullary vasa recta by angiotensin II is modulated by prostaglandin E2.

Vasa recta were dissected from outer medullary vascular bundles in the rat and perfused in vitro. Examination by transmission electron microscopy reveals them to be only outer medullary descending vasa recta (OM-DVR). To establish a method for systematic examination of vasoconstriction, OMDVR were perfused at 5 nl/min with collection pressure increased to 5 mmHg. Under these conditions, transmembrane volume flux was found to be near zero, and the transmural hydraulic pressure gradient was found to be < 15 mmHg. Over a concentration range of 10(-12) to 10(-8) M, abluminal application of angiotensin II (ANG II) caused graded focal vasoconstriction of OMDVR that is blocked by saralasin. Luminal application of ANG II over the same concentration range was much less effective. Abluminal application of prostaglandin E2 (PGE2) shifted the vasoconstrictor response of OMDVR to higher ANG II concentrations. PGE2 reversibly dilated OMDVR that had been preconstricted by ANG II. These results demonstrate that OMDVR are vasoactive segments. Their anatomical arrangement suggests that they play a key role in the regulation of total and regional blood flow to the renal medulla.

Angiotensin II↗
PubMed1994

Transport of sodium and urea in outer medullary descending vasa recta.

We dissected and perfused outer medullary vasa recta (OMVR) from vascular bundles in the rat. Permeabilities of sodium (PNa) and urea (Pu) were simultaneously determined from the lumen-to-bath efflux of 22Na and [14C]urea. PNa and Pu were also measured by in vivo microperfusion of descending (DVR) and ascending vasa recta (AVR) at the papillary tip of Munich-Wistar rats. In some OMVR PNa was indistinguishable from zero. The mean +/- SE of PNa (x 10(-5), cm/s) in OMVR was 76 +/- 9. Pu in OMVR was always very high (x 10(-5), cm/s), 360 +/- 14. There was no correlation between OMVR PNa and Pu. Inner medullary AVR and DVR had PNa of 115 +/- 10 and 75 +/- 10, respectively, and Pu of 121 +/- 10 and 76 +/- 11, respectively. PNa and Pu in papillary vasa recta were always nearly identical and highly correlated. Transport of [14C] urea in OMVR was reversibly inhibited by addition of unlabeled urea or phloretin to the bath and lumen, providing evidence for carrier-mediated transport. These data suggest that sodium and urea might traverse the wall of inner medullary vasa recta by a paracellular pathway while urea also crosses by a transcellular route in OMVR. Electron microscopic examination of seven in vitro perfused OMVR revealed no fenestrations and exposure of these vessels to 10 microM calcium ionophore A23187 or 1 nM angiotensin II resulted in reversible contraction, suggesting that in vitro perfused OMVR are DVR only.

Angiotensin II↗
PubMed1992

A model of the volumetrically-controlled hemodialysis circuit.

We developed a model that predicts the hemodynamics of the volumetrically-controlled circuit used to administer high flux hemodialysis. The equations simulate the entire blood side of the circuit so that blood and dialysate pressures can be predicted from a knowledge of circuit component and patient characteristics. An alternative method of computation has also been devised which permits measured circuit pressures to be used to predict patient blood access pressure, dialyzer resistance to flow and membrane hydraulic conductivity. Success of the model was evaluated by measuring both circuit pressure and component characteristics. The model successfully predicted circuit pressures when measured component characteristics were employed as model inputs. Conversely, the model accurately predicted circuit component characteristics when measured pressures were employed as inputs (8 patients, 30 dialyses). Specific predictions of the model include the following. Elevations of patient blood access pressure will cause blood and dialysate pressures to rise equivalently without affecting the rate of back-filtration or location of pressure equilibrium along the dialyzer axis. Elevated hematocrit is predicted to increase circuit pressures to a degree that is similar to a poorly functioning blood access, however, high hematocrit markedly augments back-filtration and moves the point of pressure equilibrium toward the dialyzer entrance. We conclude that the model provides a predictive tool that can be used to optimize circuit design. Alternatively, the model can be used to separate the influence of a poorly functioning patient access from other factors which can elevate circuit pressures.

Blood Flow Velocity↗
PubMed1992

Molecular sieving of albumin by the ascending vasa recta wall.

Molecular sieving of albumin by ascending vasa recta. Evidence exists to support the presence of an extravascular pool of albumin in the renal medullary interstitium. This study used microperfusion in vivo to measure the transport of 125I-labeled albumin from descending (DVR) and ascending vasa recta (AVR) to the papillary interstitium. Perfusions were performed during furosemide diuresis with a buffer containing FITC-labeled dextran (FITC-Dx) 2 x 10(6) mol wt and 125I-albumin. Perfusate albumin and collection pressure were adjusted to induce either zero transcapillary volume flux (Jv) or high volume flux. When Jv was zero, the collectate-to-perfusate ratios of FITC-Dx (RDX) and 125I-albumin (Ralb) in the DVR and AVR were identical implying that diffusive efflux of albumin was immeasurably small. In contrast, when Jv was increased, paired comparison of Ralb and RDX in the same AVR revealed a difference, 1.58 +/- 0.06 vs 1.72 +/- 0.08, respectively (P less than 0.01). AVR perfusions in hydropenic animals showed similar results, Ralb = 1.70 +/- 0.07 and RDX = 2.00 +/- 0.07 (P less than 0.01). These data suggest that albumin transport across vasa recta in vivo is likely to be governed by solvent drag. The reflection coefficient of the AVR wall to 125I-albumin is estimated to be 0.78.

Albumins↗
PubMed1991

Resistance of ascending vasa recta to transport of water.

A study was undertaken to determine the effect of increasing capillary pressure on volume flux in ascending vasa recta (AVR). In one experiment (group I), AVR were blocked by a single injection of paraffin wax and subjected to free-flow microperfusion at 10 nl/min. Collected fluid was obtained from the perfused vessels by micropuncture. In a second experiment (group II), AVR segments were isolated between two paraffin blocks and perfused at 10 nl/min. In group II, the collection pipette was pressurized to 0, 10, or 20 mmHg. Transmembrane volume flux was determined by measuring the change in concentration of fluorescein isothiocyanate-labeled dextran (2 x 10(6) mol wt) from perfusate to collected fluid. In group I, measurements revealed a capillary pressure of 10.3 +/- 0.5 (SE) mmHg and volume flux of 4.3 +/- 1.0 nl.mm-1.min-1. In group II, volume flux was 1.8 +/- 1.3, 5.9 +/- 1.0, and 11.2 +/- 1.1 nl.mm-1.min-1 at collection pressures of 0, 10, or 20 mmHg, respectively. Based on these data and an AVR diameter of 20 microns, AVR hydraulic conductivity is between 12.5 x 10(-6) and 18.7 x 10(-6) cm.s-1.mmHg-1. The papillary AVR have a high hydraulic conductivity. This is consistent with their role as the sole conduit for removal of water from the papillary interstitium.

Animals↗
PubMed1991

Photon-counting microcolorimeter with 40-nl cuvette.

A colorimeter with a 40-nl cuvette has been constructed. The wall of standard capillary glass was perforated to produce a sample injection port. A section of the capillary glass was drawn to a length of 1/2 cm and 80 microns ID by heating on a microforge. This produced a cuvette volume of approximately 40 nl. Two fiber-optic filaments, 80 microns in diameter, were fixed into the cuvette to transmit and receive light from the sample. The output of the colorimeter was measured with a microscope photon-counting detection assembly. It has been shown that the colorimeter enables a reduction of the plasma volume requirements of the Lowry microprotein assay from several nanoliters to 60 pl. The linearity and reproducibility of the microcolorimeter when used with the Lowry assay has been verified. The colorimeter output is several orders of magnitude above the lower limit of detection of the photon counter.

Animals↗
PubMed1991

Transport of sodium chloride and water in rat ascending vasa recta.

Experiments were undertaken to test the hypothesis that transcapillary small solute (NaCl and urea) gradients drive water across ascending vasa recta (AVR). Axial gradients of NaCl and urea were eliminated with furosemide. AVR were perfused with buffer containing fluorescein isothiocyanate-labeled dextran and 22Na. Perfusion of AVR with isotonic buffer at 10 and 20 nl/min yielded collectate-to-perfusate 22Na ratios of 0.17 +/- 0.05 and 0.34 +/- 0.03, respectively, in AVR of 601 +/- 56 and 583 +/- 46 microns mean length, respectively. A 22Na permeability of 113.2 +/- 12.8 x 10(-5) cm/s was determined. AVR were perfused at 20 nl/min with buffer NaCl of 0 (hypotonic to papilla), 161 (isotonic), or 500 mM (hypertonic). Transcapillary volume flux was not significantly different in these groups (3.8 +/- 1.5, 4.6 +/- 1.5, and 2.1 +/- 1.4 nl.min-1.mm-1, respectively). AVR were perfused in the hydropenic kidney at 5 nl/min antegrade from tip to base and retrograde from base to tip, which was a maneuver designed to impose physiological transcapillary NaCl and urea gradients of opposite direction. Volume fluxes were -1.4 +/- 0.05 and -1.3 +/- 0.04 nl.min-1.mm-1 in these groups, respectively. These data demonstrate that the AVR are highly permeable to NaCl and that physiological small solute gradients do not influence water movement across the AVR wall.

Animals↗
PubMed1991

Effect of sodium chloride gradients on water flux in rat descending vasa recta.

In the hydropenic kidney, volume efflux from descending vasa recta (DVR) occurs despite an intracapillary oncotic pressure that exceeds hydraulic pressure. That finding has been attributed to small solute gradients which may provide an additional osmotic driving force favoring water transport from DVR plasma to the papillary interstitium. To test this hypothesis, axial gradients of NaCl and urea in the papilla were eliminated by administration of furosemide and saline. DVR were then blocked with paraffin and microperfused at 10 nl/min with a buffer containing albumin, fluorescein isothiocyanate labeled dextran (FITC-Dx), 22Na, and NaCl in a concentration of 0 (hypotonic to the interstitium), 161 (isotonic) or 322 mM (hypertonic). Collectate was obtained from the perfused DVR by micropuncture and the collectate-to-perfusate ratios of FITC-Dx and 22Na were measured. A mathematical model was employed to determine DVR permeability (Ps) and reflection coefficient to NaCl (sigma NaCl). The rate of transport of water from the DVR lumen to the papillary interstitium was 2.8 +/- 0.3 (Nv = 22), -0.19 +/- 0.4 (Nv = 15), and -2.3 +/- 0.3 nl/min (Nv = 21) (mean +/- SE) when perfusate NaCl was 0, 161, or 322 mM, respectively (Nv = number of DVR perfused). The collectate-to-perfusate 22Na concentration ratios were 0.34 +/- 0.04, 0.36 +/- 0.04 and 0.37 +/- 0.03 for those groups, respectively. Based on these data, Ps is calculated to be 60.4 x 10(-5) +/- 4.0 x 10(-5) cm/s and sigma NaCl less than 0.05. The results of this study confirm that transcapillary NaCl concentrations gradients induce water movement across the wall of the DVR.

Animals↗
PubMed1990

Resistance of descending vasa recta to the transport of water.

The effect of varying intracapillary oncotic pressure on the rate of transcapillary volume flux in microperfused descending vasa recta (DVR) was studied during furosemide diuresis in the Munich-Wistar rat. At the papillary base, plasma protein concentration and hydraulic pressure were 5.7 +/- 0.1 g/dl and 11.7 +/- 0.7 mmHg in nonperfused DVR, respectively, and 5.6 +/- 0.1 g/dl and 9.4 +/- 0.4 mmHg in nonperfused ascending vasa recta (AVR), respectively. These results demonstrate that the papillary microcirculation does not remove water from the interstitium during furosemide diuresis and defines Starling forces in the pericapillary interstitium. Osmolality and urea concentration were 380 +/- 11 mosmol/kgH2O and 56 +/- 5 mM in DVR plasma at the papillary base, respectively, and 386 +/- 10 mosmol/kgH2O and 62 +/- 5 mM in DVR plasma at the tip, respectively. These results demonstrate abolition of corticomedullary small solute gradients. DVR were perfused at a rate of 10 nl/min with a buffer solution containing small-solute concentrations that matched those of plasma in nonperfused DVR. The buffer solution also contained 2 x 10(6) mol wt fluorescein isothiocyanate-labeled dextran (FITC-Dx, 5 mg/ml) and either 0.1 or 5.0 g/dl albumin. Microperfused DVR were punctured a second time downstream of the perfusion site for sample collection or servo-nulling pressure measurement. The rate of transmembrane volume flux, determined from the change in FITC-Dx concentration from perfusate to collectate, was 0.99 +/- 0.29 nl.min-1.mm-1 when perfusate contained 0.1 g/dl albumin and 0.00 +/- 0.23 nl.min-1.mm-1 with 5.0 g/dl albumin (P less than 0.01). Intracapillary hydraulic pressures were 21.7 and 20.4 mmHg during microperfusion of DVR with 0.1 and 5.0 g/dl albumin, respectively. These results demonstrate that transcapillary driving forces of 20 mmHg (5 g/dl albumin) influence transcapillary water movement across the DVR endothelium. For an average capillary diameter of 12.9 microns, DVR hydraulic conductivity is calculated to be greater than 1.4 x 10(-6) cm.s-1.mmHg-1.

Animals↗