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Biomedical subjects

J B Wade

Publications and source records attributed to J B Wade.

At least 91 records · Page 5Linked to original sources
PubMed1981

Epithelial organization and hormone sensitivity of toad urinary bladder cells in culture.

Two continuous cell lines (TB-M and TB-6c) derived from epithelial cells of the toad urinary bladder form epithelia in culture that manifest hormone-sensitive transepithelial transport. Development of transepithelial electrical resistance (R) and transport rate (ISC) are dependent on time and density of cells seeded, but steady-state ISC and R are characteristic for each cell line and independent of seeding density. Some responses of intact toad bladder are preserved in culture, whereas others are altered or absent. Neither cell line responds to vasopressin. Analogues of cAMP increase sodium transport and urea permeability in both cell lines but do not affect water permeability. The intramembrane particle aggregates associated with the vasopressin- and cAMP-induced increase in water permeability of the intact bladder could not be detected in the cell lines. Aldosterone increases sodium transport in both cell lines, and the time course and concentration dependence of the response to aldosterone are similar to those of the intact bladder. The relative effect of a series of steroids on ISC reveals corticosterone to be a more potent mineralocorticoid in cultured cells than in the intact bladder.

Animals↗
PubMed1980

Morphologic alterations in the rat medullary collecting duct following potassium depletion.

Freeze-fracture and thin-section electron microscopy and morphometry were used to characterize further the response of the rat medullary collecting duct to potassium depletion. In freeze-fracture replicas, principal cells and intercalated cells were identified based on the assumption that intercalated cells possess a high density of rod-shaped intramembrane particles in their luminal membranes. Potassium depletion caused an increase in the relative number of cells with a high density of rod-shaped particles from the control level of 22% to 31% after 2 weeks and to 36% after 4 weeks. The frequency of intercalated cells identified by thin-section criteria was, however, about 35% in controls and unchanged by potassium depletion. This suggests that intercalated cells can have two types of membrane morphology. In potassium depletion, all intercalated cells display a high density of rod-shaped particles in their luminal membranes. In addition, the luminal membrane area of intercalated cells increased more than threefold, and the density of their rod-shaped particles increased by 21%. These observations suggest that the intercalated cell and its rod-shaped particle may be involved with the potassium reabsorption that occurs in this nephron segment with potassium depletion.

Animals↗
PubMed1980

Phalloidin-induced cholestasis: a microfilament-mediated change in junctional complex permeability.

Phalloidin, administered to male rats for 7 days (500 microgram per kg/day), increased the mean hepatic content of filamentous actin. Both bile flow and bile acid excretion diminished proportionally, whereas the bile-to-plasma ratios of [3H]inulin and [14C]sucrose increased significantly from 0.08 and 0.16 in controls to 0.37 and 0.69, respectively, in phalloidin-treated animals. Simultaneously, junctional permeability was altered as noted by the free penetration of ionic lanthanum into the zonula occludens and bile canaliculus. Freeze-fracture replicas of the junctional complex revealed rearrangements of the junctional elements and regions in which only a single element separated the canaliculus from the lateral intercellular space. These findings suggest that microfilaments influence the permeability of "tight junctions" between hepatocytes and that bile constituents might reflux from the canaliculus to the intercellular space in phalloidin-induced cholestasis.

Actins↗
PubMed1979

Toad urinary bladder epithelial cells in culture: maintenance of epithelial structure, sodium transport, and response to hormones.

Epithelial cells from the toad urinary bladder have been grown in continuous culture. Many of the cells resemble the granular cell type of the urinary bladder. They form an epithelium with typical tight junctions and gap junctions. The transport properties of two cell lines have been examined. When cells of the line designated TB-M or of line TB-6c are grown on collagen-coated nucleopore filters, epithelia are formed that have transepithelial potential differences of 40 and 20 mV, resistances of 5000 and 10,000 omega-cm2, and short-circuit currents (ISC) of 8.5 and 2.5 muA/cm2, respectively. Net mucosa to serosa sodium transport accounts for all of ISC in line TB-M and for 70% of ISC in line TB-6c. Vasopressin, which stimulates adenylate cylase and ISC in the intact bladder, has no effect on the cells in culture. Cyclic AMP stimulates ISC and lowers resistance in both lines. Aldosterone stimulates ISC in both lines. This is accompanied by a fall in resistance in line TB-M and no change in resistance in line TB-6c. Amiloride inhibits ISC in TB-M cells under basal conditions and after stimulation by aldosterone. In line TB-6c amiloride has no effect under basal conditions but lowers ISC of aldosterone-treated cells to the basal level. Thus, the cells have retained the ability to form oriented, high-resistance epithelial membranes that manifest hormone-sensitive transepithelial sodium transport.

Aldosterone↗
PubMed1979

Modulation of cell membrane area in renal collecting tubules by corticosteroid hormones.

Isolated renal cortical collecting tubules obtained from rabbits treated chronically with desoxycorticosterone acetate (DOCA) have been found to possess elevated transepithelial potential differences and a greatly increased capacity for ion transport. Structural exmination of tubules from rabbits exposed to either DOCA or dexamethasone for 11--18 d reveals a marked increase in basolateral cell membrane area in these tubules. Morphometric analysis shows that this effect is specifically on the basolateral membrane area of only one of the two cell types found in this nephron segment. Increases of greater than 140% and 90% are found for the basolateral membrane area of the principal cells for DOCA and dexamethasone, respectively, but no change could be detected in the basolateral membrane area of the intercalated cells found in this nephron segment. No siginificant changes were found in luminal membrane area, cell number, or cell volume for either cell type. These observations demonstrate that significant changes in membrane area can occur in differentiated epithelia and suggest that this may be an important mechanism for modulating epithelial transport capacity.

Animals↗
PubMed1978

Membrane structural specialization of the toad urinary bladder revealed by the freeze-fracture technique. III. Location, structure and vasopressin dependence of intramembrane particle arrays.

Examination of the toad urinary bladder by freeze-fracture electron microscopy reveals intramembrane particle arrays at a number of membrane sites. An array in which particles are aggregated into closely apposed parallel rows is found in the granular cell luminal membrane of dehydrated toads fixed in situ. These aggregates are structurally indistinguishable from those previously associated with vasopressin exposure in vitro. Aggregates are not found in granular cell luminal membrane in the case of hydrated toads fixed in situ. However, structurally similar arrays are found at low frequency in the membrane of cytoplasmic vacuoles in granular cells and in the plasma membrane of basal cells in both hydrated and dehydrated toads. Aggregates are also present at these sites in control and vasopressin-treated bladders from in vitro experiments. Particle arrays characteristic of gap junctions, desmosomes and hemidesmosomes also occur in the plasma membrane of basal cells. In addition, distinctive square arrays of particles exist in the plasma membrane of the bladder's mesothelium. Although a variety of intramembrane particle arrays exist in the toad urinary bladder, only the occurrence of organized particle aggregates in the luminal membrane of granular cells appears to be associated with vasopressin exposure.

Animals↗
PubMed1977

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↗
PubMed1977

Relationship of aggregated intramembranous particles to water permeability in vasopressin-treated toad urinary bladder.

It has been previously demonstrated with freeze-fracture electron microscopy that vasopressin induces specific structural alterations of the luminal membrane of granular cells from toad urinary bladder in a dose-dependent fashion. These alterations consist of aggregated intramembranous particles and are observed both in the presence and absence of an osmotic gradient. We examined the effect of methohexital, a selective inhibitor of vasopressin-stimulated water flow, and the effect of phloretin, a selective inhibitor of urea permeability, on the structure of the granular cell luminal membrane. Methohexital treatment of the vasopressin-stimulated toad bladder reduced both the osmotic water flow and vasopressin-induced alterations of membrane structure to the same extent. Phloretin reduced urea permeability but not water flow or particle aggregation. Since neither agent affects vasopressin-stimulated sodium movement, these findings indicate that the phenomenon of particle aggregation is specifically related to vasopressin-induced water permeability and not to changes in urea or sodium permeability.

Animals↗
PubMed1976

Membrane structural specialization of the toad urinary bladder revealed by the freeze-fracture technique. II. The mitochondria-rich cell.

Examination of the toad urinary bladder by freeze-fracture electron microscopy reveals that the mitochondria-rich cells of the epithelium possess distinctive and characteristic membrane structural specialization. Unique rod-shaped intramembrane particles are found in luminal and basal membranes as well as certain intracellular vesicles of this cell type. The consistent findings of two discrete patterns of luminal membrane structural organization supports the possibility that two morphological forms of mitochondria-rich cell exist within the toad bladder epithelium.

Animals↗
PubMed1975

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.

Animals↗
PubMed1975

Membrane structural specialization of the toad urinary bladder revealed by the freeze-fracture technique. I. The granular cell.

Examination of the toad urinary bladder by freeze-fracture electron-microscopy demonstrates structural specialization of the granular cell's luminal membrane compared to its basal membrane. Although both membranes appear to possess about 1,700 intramembranous particles per mum2, those of the luminal membrane tend to be significantly larger in size. In addition, the fracturing properties of the two membranes are markedly different: the majority of particles are found on fracture face B (outer membrane face), in the case of the luminal membrane, and the majority are found on fracture face A (inner membrane face), in the case of the basal membrane. While the two fracture faces of the basal membrane possess a similar distribution of particle sizes, in the case of the luminal membrane the B face was found to possess particles generally larger than those found on the A face. It was established that the probability of luminal membrane particles adhering to face B instead of face A is closely correlated with the size of the particle. The structural specialization of the granular cell's luminal membrane may have an important relationship to the characteristic permeability properties of this membrane and the capacity of this cell type to respond physiologically to the hormone vasopressin.

Animals↗
PubMed1974

The structure of the zonula occludens. A single fibril model based on freeze-fracture.

Replicas of freeze-fractured toad urinary bladder and gallbladder were analysed in an attempt to determine the fracturing properties and structure of the zonula occludens (tight junction). Chalcroft and Bullivant have proposed that the junction has a double set of fibrils with one set associated with each of the adjacent cell membranes. However, the fracturing pattern that is observed might also result from only a single set of fibrils which is shared by the adjacent membranes if fracturing occurred around either side of the fibrils. These two models predict quite different structures at regions of the junction where tranl sitions are made between face A and face B. The relative heights of face A and face B and the shape of the transition from face A to face B do not agree with that expected according to the two fibril model but agree exactly with that expected if only a single set of fibrils existed. Further evidence for the single fibril model is derived from fractures of the mucosa membrane which cross the junction to the membrane of the adjacent cell without deflection. Such fractures reveal a single ridge which appears to be identical to the juxtaluminal fibril of the junction. In addition, small ridges are occasionally found in place of the grooves on face B which, although not consistent with the double fibril model, is expected if the single fibril model were correct. Although alternative explanations might account for these observations, we believe that the simplest and most consistent explanation is that the zonula occludens fractures as would be expected of a single set of fibrils shared by adjacent cells.

Animals↗
PubMed1974

Fracture faces of osmotically disrupted zonulae occludentes.

Exposing the mucosal epithelium of the toad urinary bladder to 240 mM urea in Ringer's solution is known to cause a dramatic increase in the permeability of the zonulae occludentes and the appearance of distended, bubble-like compartments within these junctions. Examination of such osmotically disrupted junctions with the freeze-fracture technique reveals that these bubbles result from a distention of the compartments existing within the meshwork of interconnecting fibrils characteristic of the zonulae occludentes in this epithelium. Frequent discontinuities in the meshwork of fibrils are also found after osmotic disruption of the junction. These observations indicate the essential role of these fibrils in maintaining the characteristic properties of the zonula occludens as a site of cell-to-cell attachment and as a permeability seal.

Animals↗