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  • 1
    ISSN: 1432-1424
    Keywords: Toad bladder ; Voltage-clamping ; Vasopressin ; Ouabain ; Cell volume ; X-ray microanalysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Toad urinary bladder epithelial cells were incubated in Na Ringer's with the serosal surface of the epithelium clamped at either +50 mV, O mV (short-circuited) or −50 mV with respect to the mucosal surface. Following incubation, portions of tissue were coated with an external albumin standard and rapidly frozen. Cryosections were freeze-dried and cell composition determined by x-ray microanalysis. Cell water and ion contents were unaffected when tissues were short-circuited rather than clamped close to their open-circuit potential difference (+50 mV). Incubation with vasopressin at +50 mV, and under short-circuit conditions, caused Na uptake without cell swelling or gain in Cl. Clamping at −50 mV resulted in uptake of water and ions, with considerable variation from cell to cell. These variations in cell composition were exacerbated by vasopressin. The greater the increase in water content, the greater the rise in cell Cl. However, there was no consistent pattern to the associated changes in cation contents. Most cells gained some Na. In some cells, this gain was accompanied by an increase in K. In others, the gain of Na was predominant and cell K content actually fell. At −50 mV with ouabain, many of the cells also gained water. As was found in our earlier study with ouabain under short circuit conditions (Bowler et al., 1991), there was considerable variation in the extent of the Na gain and K loss; some cells were largely depleted of K while in others the K content remained relatively normal. These results indicate differences between granular cells in the availabilities in the plasma membranes of ion pathways, either as a consequence of differences in the numbers of such pathways or in their control.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 142 (1994), S. 9-20 
    ISSN: 1432-1424
    Keywords: Cell Cl ; Cl/HCO3 exchanger ; Toad bladder ; X-ray microanalysis ; Cell volume
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Relationships between short-circuit current (I sc), cell Cl and the mechanism(s) of Cl accumulation in toad bladder epithelial cells were investigated. In serosal Cl-free gluconate Ringer, 80% of the cell Cl (measured by x-ray microanalysis) was lost over 30–60 min with an associated decrease in cell water content. Concomitantly, I sc fell to 20% of its initial value within 10 min but then recovered to 45% of its initial value despite continued Cl loss. With the reintroduction of Cl, cell Cl and I sc both recovered within 10 min. Serosal SITS (4 acetamido-4′-isothiocyano-stilbene-2,2′-disulfonate; 0.5 mm) plus bumetanide (0.1 mm), did not prevent the fall in I sc or the loss of cell Cl in gluconate medium, although they did inhibit subsequent recovery of I sc in this medium. They also prevented the recovery of I sc in Cl medium but not the reaccumulation of Cl by the cells. Although SITS and bumetanide did not prevent the loss or recovery of Cl, they modified the pattern of the ion changes. In their absence, changes in cellular Cl were twice that of the changes in measured cellular cations implicating basolateral Cl/HCO3 exchange in Cl movement. With SITS plus bumetanide present, changes of similar magnitude in Cl were associated with equivalent changes in cation, consistent with the inhibition of Cl/ HCO3 exchange.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 435 (1998), S. 528-538 
    ISSN: 1432-2013
    Keywords: Key words Crypt ; Jejunum ; Patch clamp ; K+ channel ; Cl ; channel ; Nonselective cation channel ; Ba2+ ; Forskolin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  The patch-clamp technique was used to characterise the ion channels in cells located in the mid region of mouse jejunal crypts. Six different channels were seen. A large outwardly rectified K+ channel (BK) (conductance, g at 0 mV = 92 ± 6 pS), which was highly selective for K+ [P K + (1) 〉 P Rb + (0.6) 〉〉 P Cs + (0.09) ≈ P Na + (0.07) 〉 P Li + (0.04)], had a low, voltage-independent open probability (P o) in the on-cell (O/C) configuration and appeared in 66% of the patches. In inside-out (I/O) patches, this channel had a linear current/voltage (I/V) relationship (g = 132 ± 3 pS), P o was voltage dependent and it was blocked by cytoplasmic Ba2+ (5 mmol/l). An intermediate K+ channel (IK) which was present in 49% of O/C patches, had a linear I/V (g = 38 ± 3 pS), ran-down in O/C patches, and was not seen in I/O patches. A number of smaller channels (SC) with conductances ranging from 5 to 20 pS were seen in 16% of O/C patches. Also present in the basolateral membrane were a Cl– channel (ICOR) and a nonselective cation channel (NSCC). These channels were only seen in I/O patches. ICOR had an outwardly rectified conductance (g at 0 mV = 36 ± 2 pS), its P o was independent of voltage and unaffected by variations in cytoplasmic Ca2+ (100 nmol/l to 1 mmol/l) or ATP (0–1 mmol/l). The NSCC had a linear conductance (20 ± 1 pS), its P o increased with depolarisation and elevation of cytoplasmic [Ca2+] (≥ 10 μmol/l), but was reduced by cytoplasmic ATP. None of the basolateral channels described here were activated by cAMP-dependent secretagogues, although a Cl– conductance was activated. This cAMP-dependent Cl– conductance was distinct from the basolateral Cl– channel and thus is most likely located in the apical membrane.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-136X
    Keywords: Key words Mannitol ; Fluorescein ; Permeability ; Salmon ; Posterior intestine ; Enhanced epithelial permeability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract This study characterised the permeability of the salmonid posterior intestine in vivo, to two hydrophilic markers of different molecular weight, both in the presence and absence of sodium deoxycholate (SDA), and determined the influence of mucosal secretions. The posterior intestine of chinook salmon was cannulated with a balloon catheter and the lumen infused with a solution of fluorescein and 14C-mannitol. In treated fish, the solution also contained 5.0 mmol · l−1 SDA. Blood samples from the dorsal aorta were taken at regular time intervals over 3 h. Clearances and volumes of distribution were assessed by intravenous administration of the markers to another group of fish. In the absence of SDA, low permeabilities were recorded for both markers; however, permeabilities for both were significantly greater in the treated groups. Both solutes had volumes of distribution similar to values reported elsewhere. Metabolism of fluorescein by the liver resulted in its plasma clearance. In contrast, elimination of mannitol was negligible during the study period, probably due to the lowered glomerular filtration rates observed in sea water adapted fish. Compared to in vitro investigations, in vivo mucus secretions were significantly lower and solute delivery across the epithelium was higher. Results from these in vivo investigations have implications for the oral delivery of peptides to salmonids.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-136X
    Keywords: Key words Proximal and distal intestine ; Mannitol flux ; Transepithelial electrical resistance ; Enhancement ; Salmon
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The objective of this study was to assess regional variations in the permeability of the salmon posterior intestine and to evaluate the effect of permeability enhancers as a basis for oral delivery of biologically active peptides. Proximal and distal portions of the posterior intestine of the chinook salmon (Oncorhynchus tshawytscha) were removed, mounted as flat sheets in Ussing chambers and superfused with trout Ringer's. Intestinal permeability was assessed under short-circuit conditions by measurement of 14C-mannitol (mucosal to serosal) flux. Tissues were treated either with the mucolytic agent dithiothreitol (10 mmol · l−1), the permeability enhancer sodium deoxycholate (5.0 mmol · l−1) or both and compared to untreated controls. Both proximal and distal control tissues had low permeabilities, but the distal region had a lower transepithelial electrical resistance and produced significantly less mucus. Treatment with either dithiothreitol or sodium deoxycholate alone reduced mucus adhering to tissue in both regions but did not increase permeability or change transepithelial electrical resistance. In the distal region, sequential treatment with both agents significantly reduced adhering mucus, decreased transepithelial electrical resistance, and increased tissue permeability. The salmon posterior intestine can be divided into proximal and distal regions. The distal region is more likely to have the necessary permeability and responsiveness to enhancement for the successful delivery of peptides or polar drugs.
    Type of Medium: Electronic Resource
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