Digit symbol performance in mild dementia and depression.
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Aged↗
Biomedical subjects
J B Wade
Publications and source records attributed to J B Wade.
Effect of sleep deprivation on first-year residents' response times, memory, and mood.
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Affect↗
Water permeability and particle aggregates in ADH-, cAMP-, and forskolin-treated toad bladder.
Osmotic water flow was used to evaluate total tissue water permeability (Ptissue), and luminal membrane particle aggregates, presumed sites for transmembrane water movement, were quantified to assess luminal membrane water permeability, in bladders treated with maximally stimulating concentrations of antidiuretic hormone (ADH), adenosine 3',5'-cyclic monophosphate (cAMP), and forskolin. Aggregates were as numerous and occupied the same fractional area of the luminal membrane in response to cAMP treatment (10 mM) as treatment with ADH (20 mU/ml). Ptissue in cAMP-treated tissues, however, was only half of that induced by ADH (P less than 0.001). A similar disparity in the relationship between aggregates and Ptissue occurred for additional bladders treated with 50 microM forskolin, which is known to increase endogenous cAMP to levels much greater than caused by maximally stimulating concentrations of ADH. Although Ptissue achieved with forskolin was the same in paired bladders treated with ADH, aggregates were far more numerous (P less than 0.05) and occupied much more membrane area (P less than 0.05) with forskolin. These observations are consistent with the view that aggregate appearance in the luminal membrane is a function of intracellular cAMP. The finding that the hydrosmotic response of toad bladder to both cAMP and forskolin compared with ADH stimulation is reduced relative to measured changes in luminal membrane aggregates suggests that the effect of ADH in altering water permeability involves additional regulation via a non-cAMP-mediated mechanism. This latter event would appear to be by an effect of ADH on the permeability of a resistance at a postluminal membrane site and/or possibly on the permeability of aggregates in the luminal membrane.
Animals↗
Visualization of endocytosed markers in freeze-fracture studies of toad urinary bladder.
Antidiuretic hormone (ADH) stimulation increases the apical membrane water permeability of granular cells in toad urinary bladder. This response correlates closely with the fusion of tubular cytoplasmic vesicles with the membrane and delivery of intramembrane particle (IMP) aggregates from the tubules (aggrephores) to the apical membrane. These aggregates are believed to be associated with the channels responsible for the water permeability increase. Removal of ADH triggers apical membrane endocytosis and disappearance of aggregates from the apical membrane. However, it has been unclear whether aggregate disappearance is due to disassembly of aggregates within the apical membrane or to their endocytic retrieval as intact structures. Using colloidal gold and horseradish peroxidase to follow endocytosis from the apical surface after ADH removal, we have directly observed in cross-fractured bladder cells the intramembrane structure of intracellular vesicles that contain these fluid-phase markers. Under these conditions, intact aggregates can be identified in the membrane of tubular endocytosed vesicles. This directly demonstrates that conditions which lower apical membrane water permeability cause the tubular aggrephores to "shuttle" intact aggregates from the apical membrane back into the cytoplasm. An additional population of vesicles with tracer are found which are spherical and display structural features of the apical membrane, as well as occasional aggregates. These vesicles may be responsible for retrieval of aggregates from the surface apical membrane.
Animals↗
Neuropsychological implications of metrizamide myelography.
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Humans↗
Role of membrane fusion in hormonal regulation of epithelial transport.
Fusion of cytoplasmic vesicles with the plasma membrane represents an important mechanism whereby additional transport proteins can be introduced into the cell membrane. Although it is apparent that some epithelial hormone responses, such as the vasopressin-induced change in water permeability, utilize this mechanism, the mechanism of some other responses remains uncertain. In addition, many details concerning the intracellular systems responsible for controlling vesicle translocation and membrane fusion in epithelial hormone responses remain to be elucidated.
Aldosterone↗
Fluorescent markers to study membrane retrieval in antidiuretic hormone-treated toad urinary bladder.
Antidiuretic hormone (ADH) stimulation of toad urinary bladder causes fusion of intracellular vesicles called aggrephores with the apical plasma membrane of granular cells. Aggrephores contain intramembrane particle aggregates whose appearance in the apical membrane is believed to produce a large increase in its water permeability. ADH removal (ADH washout) is thought to cause the retrieval of aggrephores into granular cell cytoplasm. We studied granular cell uptake of dextran and horseradish peroxidase conjugated with fluorescein, rhodamine, or both during ADH washout. Granular cell uptake of fluorescent dextran was dependent on prior exposure to ADH, a linear function of dextran concentration, and increased by a transepithelial osmotic gradient. Immediately after removal of ADH, granular cell fluorescence was finely dispersed and located near the apical surface. Subsequently, it coalesced into larger bodies. This change was most apparent when a single bladder was subjected to two cycles of ADH stimulation and removal using a dextran containing a different fluorophore for each cycle. The ultrastructural correlate for these fluorescent patterns was identified using rhodamine-labeled horseradish peroxidase. Electron microscopy showed that after detachment from the apical membrane, label was initially in tubular-shaped vesicles near the apical surface. Later, these vesicles clustered near multivesicular bodies and transferred their label to these structures. These tubular vesicles closely resemble the morphology of aggrephores visualized by freeze-fracture electron microscopy. We conclude that these fluorescent compounds can be used as markers for the luminal contents of membrane retrieved during ADH washout and allow detailed study of its intracellular processing.
Animals↗
Evaluation of granule exocytosis in toad urinary bladder.
Addition of phorbol myristate acetate (PMA) to the mucosal bathing solution induces exocytosis of the granules found in the apical cytoplasm of toad urinary bladder epithelial cells. Because mucosal application of PMA also increases epithelial water permeability in the complete absence of vasopressin, these observations have suggested that granules might have a role in mediating the permeability response to vasopressin. From electron microscopic immunolocalization studies we have identified a 70-kDa protein as a component of the apical surface glycocalyx that is stored in the granules. Using antibodies to this protein in a competitive immunoassay, we found that addition of PMA to the mucosal side results in more than a fourfold increase in antigen release into the mucosal bath. The amount of antigen detectable on the mucosal surface of the cells was doubled by mucosal PMA exposure. Serosal application of PMA or addition of the inactive analogue phorbol didecanoate to either surface failed to produce significant antigen release. These results are consistent with electron microscopic studies showing exocytosis of granules only with mucosal exposure to PMA. However, immunoassay of granule antigen after vasopressin stimulation of water permeability failed to show a detectable change in granule exocytosis. We conclude that granule exocytosis does not play an important role in mediating vasopressin-induced changes in permeability.
Animals↗
Transepithelial water flow regulates apical membrane retrieval in antidiuretic hormone-stimulated toad urinary bladder.
Antidiuretic hormone (ADH) increases the osmotic water permeability (Posm) of toad urinary bladder. This increase is believed to be produced by fusion of intracellular vesicles called aggrephores with the granular cell apical plasma membrane. Aggrephores contain intramembrane particle aggregates postulated to be water channels. ADH-stimulated Posm is decreased by osmotic gradient exposure, which is termed flux inhibition. We studied flux inhibition by exposing ADH-stimulated bladders to various osmotic gradients. Osmotic water flow was initially proportional to the applied osmotic gradient, but Posm decreased with time. Ultrastructural and quantitative studies of endocytosis demonstrate that apical membrane retrieval was a direct function of the transepithelial osmotic gradient. Posm remained unchanged when apical membrane retrieval was blocked by incubation of bladders at 2 degrees C, or under low water-flow conditions. These effects were reversed by increases in temperature or the applied osmotic gradient. We conclude that apical membrane retrieval causes the phenomenon of flux inhibition.
Animals↗
Structural adaptation in initial collecting tubule following reduction in renal mass.
Tubular function studies have shown that nephrectomy leads to a sharp increase in potassium secretion by initial collecting tubules. The aim of this study was to examine the relationship between potassium transport and the cellular morphology of the distal nephron. Clearance and ultrastructural studies were conducted in rats 10 days after 75% nephrectomy. Fractional urinary potassium excretion increased from 23.4 +/- 3.8% in controls to 63.9 +/- 10.1% in nephrectomized animals. Stereological analysis of principal cells in the initial collecting tubule revealed that following nephrectomy the surface density of the basolateral membrane increased from 2.32 +/- 0.12 to 4.11 +/- 0.20 micron2/micron3. While the basolateral membrane surface density of intercalated cells in this segment was not affected by nephrectomy, the luminal membrane surface density increased from 0.29 +/- 0.03 to 0.54 +/- 0.08 micron2/micron3. Ultrastructural changes were not observed in cells of the distal convoluted tubule. These observations demonstrate that cell type specific changes in membrane surface density accompany the adaptive increase in ion transport that occurs with a reduction in renal mass. The enhanced potassium excretion associated with nephrectomy may be partly mediated by a specific increase in the basolateral membrane of principal cells. The increase in luminal membrane of intercalated cells may be related to the increase in acid transport per surviving nephron following nephrectomy.
Animals↗
Membrane structural studies of the action of vasopressin.
Freeze-fracture electron microscopy of the toad urinary bladder indicates that distinctive intramembrane particle aggregates are responsible for the increase in apical membrane water permeability that occurs with vasopressin (VP) stimulation. In unstimulated bladders the aggregates occur in the cytoplasm of the cells in tubular membrane structures now called aggrephores. After stimulation by VP, aggrephores are shuttled to the surface and fuse with the apical membrane. It is suggested by structural observations and by measurements of membrane capacitance that the area of aggregates inserted into the apical membrane is much greater than previously suspected because many aggregates remain in the wall of the fused aggrephores. The area of the aggregates in a stimulated bladder is sufficiently large for these structures to represent an organized array of water channels that mediates the change in apical membrane permeability. Work with antibodies supports the concept that these channels are not always resident in the apical membrane but become inserted only after stimulation by the hormone VP.
Animals↗
Interhemispheric transfer in schizophrenia and affective disorder.
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Adult↗
Ultrastructural characterization of cholesterol distribution in toad bladder using filipin.
The polyene antibiotic filipin was used to characterize the cholesterol distribution in the membranes of the toad bladder epithelium in freeze-fracture replicas. The apical membranes of granular and mitochondria-rich cells incorporate moderate amounts of filipin while the basolateral membranes of both cell types incorporate substantially greater amounts. Intracellular membranes, in general, take up very little filipin. The major exception to this is the granule membrane, which appears to be rich in cholesterol. An inverse correlation was found between the density of filipin-sterol complexes in the apical membrane and the incidence of granules in the cytoplasm. This suggests that fusion of granules with the apical membrane may be responsible for variation in the concentration of cholesterol in the apical membrane. Thirty minutes following vasopressin exposure, there is no consistent change in the cholesterol content of the apical membrane of granular cells as measured by the incidence of filipin-sterol complexes. The lack of change in the amount of membrane cholesterol indicates that the vasopressin-induced increase in transepithelial water permeability is not mediated by a change in cholesterol content of the apical membrane.
Animals↗
Structural integrity of hepatocyte tight junctions.
The significance of discontinuities frequently found in freeze-fracture replicas of the tight junction was evaluated using complementary replicas of hepatocyte junctions from control and bile duct-ligated rats. An extensive analysis of complementary replicas using rotary platinum shadowing indicates that discontinuities in the protoplasmic (P) fracture face do not represent structural breaks in the tight-junctional network. In no case did P-face discontinuities correspond with interruptions in the groove network on the complementary extracellular (E) face. Quantitative analysis of replicas shows that P-face discontinuities result in part from "transfer" of material to the complementary E face (approximately 7% of the junctional length). However, many P-face discontinuities (7-30% of the junctional length) are matched only by a groove on the complementary E face. This finding demonstrates that a significant amount of material can be lost during freeze-fracture. An analysis of junctions from bile duct-ligated rats, which are known to have an increased paracellular permeability, shows comparable transfer and loss of material. However, the number of junctional elements and the tight-junction network density was significantly reduced by bile duct ligation. These observations indicate that discontinuities in tight-junctional elements result during the preparation of freeze-fracture replicas and are not physiologically important features of the junctional barrier. Variation in the number of elements provides the best explanation for observed differences in tight-junction permeability.
Animals↗
Evaluation by capacitance measurements of antidiuretic hormone induced membrane area changes in toad bladder.
A technique for estimating effective transepithelial capacitance in vitro was used to investigate changes in epithelial cell membrane area in response to antidiuretic hormone (ADH) exposure in toad bladder. The results indicate that transepithelial capacitance increases by about 30% within 30 min after serosal ADH addition and decreases with ADH removal. This capacitance change is not blocked by amiloride and occurs whether or not there is a transepithelial osmotic gradient. It is blocked by methohexital, a drug which specifically inhibits the hydro-osmotic response of toad bladder to ADH. We conclude that the hydro-osmotic response of toad bladder to ADH is accompanied by addition of membrane to the plasmalemma of epithelial cells. This new membrane may contain channels that are permeable to water. Stimulation of Na+ transport by ADH is not related to membrane area changes, but appears to reflect activation of Na+ channels already present in the cell membrane before ADH challenge.
Amiloride↗
Binding of peanut lectin to specific epithelial cell types in kidney.
The binding of peanut agglutinin (PNA) to epithelial membranes of the rabbit kidney was evaluated at the light- and electron-microscope level using PNA conjugated to horseradish peroxidase. In the renal cortex and outer stripe of the medulla PNA appears to bind exclusively to the luminal membrane of intercalated cells in connecting tubules and collecting ducts. PNA also binds to the thin descending limb of the loop of Henle in the inner stripe and inner zone of the medulla. This very specific affinity of PNA should be useful in the isolation and characterization of specific cell types in cytologically heterogeneous epithelia.
Animals↗
Structure of tight junctions during Cl secretion in the perfused rectal gland of the dogfish shark.
In epithelia that secrete sodium chloride, high-conductance tight junctions between cells have been proposed as the primary pathway for transepithelial sodium flux. We examined the properties of tight junctions in the perfused rectal gland of the dogfish shark during basal secretion and following adenosine 3',5'-cyclic monophosphate stimulation of sodium chloride secretion. Freeze-fracture electron microscopy revealed extensive interdigitation of adjacent cells with an associated amplification in the length of tight junctions per area of luminal surface, averaging 102 +/- 4.7 m/cm2 in outer regions of 80 +/- 6.7 in inner regions of the gland. Marked heterogeneity of junctional structure was present with junctional elements varying from single strands to three duplex elements and junctional depth varying from 15 to 60 nm. In glands perfused with lanthanum chloride, ionic lanthanum filled the intercellular space up to but not through the tight junctions. Characteristics of tight junctions were not different during basal and maximally stimulated sodium chloride secretion. These studies define tight junctions in the rectal gland as an anatomical barrier capable of restricting the passage of relatively small molecules such as urea while providing a greatly amplified junctional area for the passive diffusion of sodium and water.
Animals↗
Structural and functional study of the rat distal nephron: effects of potassium adaptation and depletion.
To examine the relationship between tubular transport of potassium and cell structure in segments of the superficial distal nephron, we performed potassium transport and quantitative electron microscopic studies in rats after potassium adaptation and potassium depletion. In distal nephrons continuously microperfused in vivo, potassium adaptation stimulated potassium secretion by 200%. Microperfused distal convoluted tubules (earliest portion of accessible distal nephron) did not, however, secrete potassium in potassium adapted animals. Morphometric analysis of the distal convoluted tubule also revealed no detectable effect of potassium diet on the structure of the distal cell type. In contrast, examination of the connecting tubule and the initial collecting tubule of the distal nephron demonstrated a striking increase in basolateral membrane in potassium-adapted animals. This change was limited to the connecting tubule cell and the principal cell type. No structural change of the intercalated cell type in either segment was associated with altered potassium transport. We conclude that cells of the distal convoluted tubule do not secrete potassium. Functional and morphologic evidence suggests that potassium is secreted by the connecting tubule cell and the principal cell of the connecting tubule and the initial collecting tubule, respectively.
Animals↗