Vasopressin: induced structural change in toad bladder luminal membrane. 1975.
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Animals↗
Biomedical subjects
V A DiScala
Publications and source records attributed to V A DiScala.
Effects of serosal hypertonicity on water permeability in toad urinary bladder.
We studied in toad urinary bladder the effects of serosal hypertonicity on tissue water permeability, granular cell luminal membrane water permeability, and granular cell luminal membrane particle aggregates and compared them with effects of antidiuretic hormone (ADH). In tissues challenged by a hypertonic (447 mosmol/kgH2O) serosal bath, luminal membrane aggregates were structurally similar to those caused by ADH. The tissue water permeability increase induced by serosal hypertonicity was much less than that caused by a maximally stimulating concentration of ADH on tissue in isotonic serosal baths with approximately the same transmural gradient. The difference is explained not only by a reduced incidence of luminal membrane aggregates but also by an increased resistance to water movement at a postluminal membrane site. Measurements of luminal membrane water permeability showed a close correlation with luminal membrane aggregate frequency, indicating that the calculated permeability of an individual aggregate was a constant. Thus the relation of luminal membrane aggregates to tissue osmotic permeability is modified by serosal hypertonicity. Morphological examination of these tissues suggested that luminal membrane aggregates may be less stable in the absence of hormone. This was evident by the proportionally greater number of structures interpreted as aggregates captured in the process of disassembly ("patches"). Membrane depressions containing intramembrane particles ("craters") were also observed. They corresponded in terms of frequency and size to coated pits as seen in thin sections.
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Variability of cellular responsiveness to ADH stimulation in toad urinary bladder.
The hydrosmotic response of toad bladder to antidiuretic hormone (ADH) is quantitatively linked to the induced fusion of aggrephores with, and the appearance of aggregates of tightly packed intramembrane particles in the luminal membrane of granular cells. We used these morphological indexes of hormonally induced cell activation 1) to assess the variability of individual cell responsiveness to a maximally stimulating concentration of ADH and 2) to compare cell response patterns in paired tissues where the extent of whole tissue stimulation, as evidenced by transtissue water flow, was either maximal or submaximal. The results indicate that individual cell responsiveness within the same tissue to standardized maximal ADH treatment varies between two- and sevenfold, depending on the morphological endpoint measured. Furthermore, based on skewness in endpoint distribution, this variability appears to reflect inherent heterogeneity of granular cell reactivity to hormone. In relation to proportional tissue responses elicited by different stimulating concentrations of ADH, our observations of luminal membrane aggregate incidence suggest that the responding cells, whatever their sensitivity, participate in a graded, rather than "all-or-none," "on-off" manner.
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Effect of hydrostatic pressure on ADH induced osmotic water flow in toad bladder.
The effect of hydrostatic pressure (HP) on antidiuretic hormone (ADH) stimulated osmotic water flow (Jv) across the toad urinary bladder was evaluated. Jv for ADH-stimulated bladders was significantly reduced by an elevation of the serosal HP gradient to 1 cm H2O. Subsequent elimination of the HP gradient resulted in a recovery of Jv. Serosal HP also caused a reversible increase in sucrose permeability (P sucrose). For ADH-treated bladders fixed with glutaraldehyde during serosal HP exposure, subsequent exposure to a mucosal or serosal HP gradient caused acceleration or inhibition of Jv, respectively. The reduction in ADH-associated Jv with serosal HP was apparently caused by a back-flux of water through a paracellular pathway. Jv and P sucrose were not affected by mucosal HP during ADH stimulation. The results suggest a specific sensitivity of a paracellular pathway to a small serosal HP gradient in bladders with ADH-stimulated water flow. The reversibility of this effect on P sucrose suggests that the elements comprising the apical junctions are dynamic structures capable of recovering at least some of their permeability properties.
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Renal tubular sodium reabsorption and oxygen consumption in the hypothyroid dog.
The degree to which an insufficiency of thyroid hormone (TH) affects renal tubular sodium reabsorption (RNa) and oxygen consumption (QO2) in vivo, was investigated in a mature mammalian kidney. The results suggest that TH influences renal QO2 as a result of alterations in renal hemodynamics and the renal tubular handling of sodium. Changes in GFR played a major role in the regulation of RNa and QO2. This is suggested by the comparison of hypothyroid animals with and without renal compensatory hypertrophy. In the latter group a twofold lower GFR was associated with parallel changes in RNa and QO2. However, in going from euthyroid (E) to hypothyroid (H) conditions, the decrease in RNa of 31 +/- 4% was greater than the reduction in GFR of 25 +/- 3% (p less than .05). Thus, a decrease in RNa of 6 +- 2% under hypothyroid conditions could not be attributed to the changes in GFR. The normal renal stoichiometry between RNa and QO2 in hypothyroid animals indicates that the above changes in RNa were accompanied by comparable changes in QO2. The decrease in RNa in hypothyroid animals was not associated with a significant change in kidney weight (E:48.2 +/- 4.0gm., n = 5 and H:45.4 +/- 1.4gm, n = 5; pN.S.) The results suggest that an adaptive response to changes in GFR is the predominant cause of the decrease in RNa and renal QO2 under hypothyroid conditions. A modest reduction in renal tubular sodium reabsorptive capacity in hypothyroid animals is probably secondary to a direct effect of TH on active sodium transport.
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Effects of chronic sodium depletion on renal tubular sodium and water reabsorption in man.
In order to assess the effects of sodium depletion on tubular Na and H2O transport in man, renal clearance studies were performed under control conditions and after sodium depletion in 7 normal subjects. The control diet contained 250 mEq of Na daily. Sodium depletion was induced by placing subjects on a 20-mEq/day na diet and administering furosemide (40 mg) dialy for 3 days. Balance studies revealed that the subjects were in Na balance under control conditions, and that after sodium depletion had been established they were in a sodium-retaining state. Subjects were studied under both conditions, after H2O hydration and during a subsequent hypotonic mannitol infusion. After H2O hydration the minimal urinary osmolality averaged 61 and 67 mosm/kg in the sodium depletion and control states, respectively. Sodium depletion was associated with a significant reduction in C inulin, sodium clearance (CNa), urine flow (V), fractional urine flow (V/GFR), free water (CH2O), and fractional free water clearance (CH2O/GFR). During hypotonic mannitol infusion indices of absolute and fractional delivery of Na from the proximal tubule (CH2O + CNa; CH2O/GFR + CNa/GFR) were consistently reduced after sodium depletion and rose more slowly as absolute and fractional distal sodium chloride supply (CH2O + CNa; CH2O/GFR + CNa/GFR) were increased. These data are consistent with the view that modest sodium depletion in man provokes an increase in fractional sodium chloride reabsorption in both the proximal and distal tubules.
Absorption↗
Relation of ADH effects to altered membrane fluidity in toad urinary bladder.
Membrane fluidity, urea permeability, and osmotic water permeability in toad urinary bladder are regularly enhanced by antidiuretic hormone (ADH). In addition, organized intramembranous particle aggregates, which correlate specifically with hormonally stimulated water permeability, are found in granular cell luminal membranes consequent to ADH stimulation. In this investigation ADH-stimulated changes in urea and osmotic water permeability and luminal membrane aggregates at room temperature (24.8 +/- 0.4 degrees C) and in the cold 10.6 +/- 0.2 degrees) were compared with corresponding changes in membrane fluidity, as assessed by n-butyramide permeability. Although a critical level of membrane fluidity is undoubtedly required, the occurrence of aggregates in the luminal membrane is independent of an accompanying hormonally induced change of membrane fluidity. ADH-stimulated osmotic water permeability in toad bladder is also independent of the coincident change in membrane fluidity, and as a process almost certainly involves membrane channels, not a solubility-diffusion process through membrane lipids. For ADH-stimulated transbladder urea movement, channels seem to be involved as well, and the change induced in membrane fluidity by ADH could be an underlying factor in their formation.
Animals↗
Effect of osmotic gradient on ADH-induced intramembranous particle aggregates in toad bladder.
Paired toad urinary bladders were prepared without or with an osmotic gradient (175 mosM) across them, stimulated for 2.5 (n = 6), 5 (n = 6), 30 (n = 6) or 60 (n = 6) min with ADH (20 mU/ml), and studied by freeze-fracture electron microscopy. Water permeability at these times was assessed in additional bladders (n = 6 for each case) after tissue fixation according to the technique of Eggena. After both 60 and 30 min of ADH stimulation, the presence of a gradient compared with the absence of one was associated with fewer aggregates (242 +/- 35 vs. 382 +/- 14 x 235 micron-2 at 60 min, P less than 0.01; 279 +/- 36 vs. 470 +/- 51 x 235 micron-2 at 30 min, P less than 0.01) and lower water permeability (8.4 +/- 1.1 vs. 18.8 +/- 1.8 microgram x min-1 x cm-1 x mosM-1 at 60 min, P less than 0.005; 9.2 +/- 1.0 vs. 22.0 +/- 2.1 microgram x min-1 x cm-2 x mosM-1 at 30 min, P less than 0.001). In addition, with a gradient both maximum water permeability and maximum aggregate frequency were reached nearly together; a similar correspondence occurred without a gradient. We conclude that in the presence of an osmotic gradient both the ADH-associated aggregates and the water permeability response to ADH are prevented from reaching the higher levels observed in bladders not exposed to a gradient.
Animals↗
Evidence that ADH-stimulated intramembrane particle aggregates are transferred from cytoplasmic to luminal membranes in toad bladder epithelial cells.
In freeze-fracture (FF) preparations of ADH-stimulated toad urinary bladder, characteristic intramembrane particle (IMP) aggregates are seen on the protoplasmic (P) face of the luminal membrane of granular cells while complementary parallel grooves are found on the exoplasmic (E) face. These IMP aggregates specifically correlate with ADH-induced changes in water permeability. Tubular cytoplasmic structures whose membranes contain IMP aggregates which look identical to the IMP aggregates in the luminal membrane have also been described in granular cells from unstimulated and ADH-stimulated bladders. The diameter of these cytoplasmic structures (0.11 +/- 0.004 micrometers) corresponds to that of tubular invaginations of the luminal membrane seen in thin sections of ADH-treated bladders (0.13 +/- 0.005 micrometers). Continuity between the membranes of these cytoplasmic structures (which are not granules) and the luminal membrane has been directly observed in favorable cross-fractures. In FF preparations of the luminal membrane, these apparent fusion events are seen as round, ice-filled invaginations (0.13 +/- 0.01 micrometer Diam), of which about half have the characteristic ADH-associated aggregates near the point of membrane fusion. They are less numerous than, but linearly related to, the number of aggregates counted in the same preparations (n = 78, r = 0.71, P less than 0.01). These observations suggest that the IMP aggregates seen in luminal membrane after ADH stimulation are transferred preformed by fusion of cytoplasmic with luminal membrane.
Animals↗
Vasopressin and collecting duct intramembranous particle clusters: a dose-response relationship.
Vasopressin increases the permeability of collecting ducts to water. Administration of this hormone is also associated with an increase in intramembranous particle clusters in rat collecting duct luminal membrane (CDLM) as revealed by freeze-fracture electron microscopy. To determine whether this morphologic alteration of CDLM is quantitatively related to the dose of vasopressin, anesthetized Brattleboro homozygous rats were given the hormone at different doses. CDLM from kidneys removed before and during infusion were examined by freeze-fracture electron microscopy. The frequency of CDLM clusters as well as the area of membrane occupied by them was related to the dose of vasopressin. In a separate experimental protocol, a decrease in intramembranous particle clusters accompanied a decrease in urinary osmolality when vasopressin was stopped. We conclude that CDLM intramembranous particle clusters represent a specific structural change related to the action of vasopressin. Accordingly, quantitation of CDLM clusters may serve as an end point for the study of vasopressin-induced water permeability.
Animals↗
Vitamin A supplements in hemodialysis patients.
A total of 14 hemodialysis patients were followed for an average of 8.4 months after stopping oral vitamin A supplements (5,000 U daily). No change occurred in the mean vitamin A plasma levels. 11 of the 14 patients were followed for an average of 16.3 months, and no change was noted in the mean vitamin A level. Vitamin A intake does not seem to effect the plasma vitamin A level in hemodialysis patients. This is consistent with the possibility that the vitamin A levels in hemodialysis patients are increased because of an increase in retinol binding protein.
Adult↗
Temperature dependence of ADH-induced water flow and intramembranous particle aggregates in toad bladder.
Antidiuretic hormone (ADH)-induced luminal intramembranous particle aggregates and hormonally stimulated water flow in toad urinary bladder are reduced simultaneously with a reduction in temperature. When water movement is factored by the aggregation response, the apparent activation energy for this process decreases from 12.1 +/- 1.6 to 3.0 +/- 2.3 kilocalories per mole. The data are consistent with the view that the particle aggregates contain sites for transmembrane water movement and that these sites behave as pores.
Animals↗
Time course of ADH-induced intramembranous particle aggregation in toad urinary bladder.
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Animals↗
Antidiuretic hormone-induced intramembranous alterations in mammalian collecting ducts.
Freeze-fracture electron microscopy had previously revealed antidiuretic hormone-induced aggregates of intramembranous particles in amphibian urinary bladder. To investigate the effects of antidiuretic hormone (ADH) in another ADH-sensitive epithelium, namely, mammalian renal collecting ducts, freeze-fracture studies were carried out in Brattleboro homozygous rats. Collecting duct luminal membranes of ADH-treated homozygotes showed intramembranous particle clusters (117 +/- 17/100 micron2) that were loosely packed and that occurred on both exoplasmic (E) and protoplasmic (P) faces. Untreated, control homozygous rats had significantly less (3 +/- 1/100 micron2) clusters. Changes similar to those seen in ADH-treated rats were observed in water-deprived Wistar rats. The clustered particles differed from those seen in ADH-treated amphibian urinary bladder in that the latter occurred only on the P face and were more densely packed. Nevertheless, our observations suggest a common membrane effect for ADH action that may apply in mammals and amphibia alike.
Animals↗
Membrane structural and functional responses to vasopressin in toad bladder.
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Animals↗
Freeze-fracture electron microscopy: relationship of membrane structural features to transport physiology.
Recent observations utilizing freeze-fracturing electron microscopy are discussed which indicate that the membrane structural features visualized by this technique may in some instances be related to specialized membrane transport properties. The occurrence of organized aggregates of intramembrane particles observed in vasopressin or cAMP-treated toad urinary bladder has been found to be closely correlated with induced changes in the permeability of the luminal membrane. Although a cautious interpretation is considered appropriate, these observations raise the possibility that some aspects of hormone action may be restricted to limited regions of membrane. Difficulties in interpretation and some serious limitations of the freeze-fracture technique are discussed.
Animals↗
Calcium homeostasis during hypertonic volume expansion in the hypothyroid rat.
The clearance characteristics of calcium and the relationship of coupled Na+ and Ca++ renal transport were evaluated in the hypothyroid rat (TX) during volume expansion with 1.5% saline. Despite similar hydropenic plasma calcium, after 11 ml/100 g body wt of infusion volume, the plasma calcium was significantly lower in the TX group (TX, 6.1 +/- 0.2; TI, 7.0 +/- 0.1; P less than .001). With continued infusion, plasma calcium in the TX group continued to drop to 5.3 +/- 0.3 while in the control rats (TI group) it rose to 8.0 +/- 0.1. This marked hypocalcemia in the TX group could not be explained by urinary losses of calcium, hemodilution, deficient parathyroid response, or changes in ultrafilterable plasma calcium. The higher fractional calcium excretion found in the TX group paralleled the higher fractional sodium excretion, and the coupling between calcium and sodium excretion was preserved in hypothyroidism under these conditions. Our results indicate an abnormal calcium homeostasis in the hypothyroid rat. The data do not support a renal cause for this disturbance but are consistent with the view that mobilization of calcium from the bone is sluggish in hypothyroidism during volume expansion.
Animals↗
Vasopressin: induced structural change in toad bladder luminal membrane.
Freeze-fracture electron microscopy demonstrates that vasopressin stimulation of isolated toad bladder alters the structure of the luminal membrane of granular cells. This alteration consists of an ordered aggression of intramembranous particles, and appears to be of functional significance, since the frequency of aggregation sites per area of membrane is closely correlated with vasopressin-induced osmotic water flow.
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