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Gluconeogenesis in toad urinary bladder 总被引:2,自引:0,他引:2
A D Morrison D B Goodman H Rasmussen A I Winegrad 《Biochimica et biophysica acta》1972,273(1):122-131
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The kinetics and distribution of potassium in the toad bladder 总被引:4,自引:0,他引:4
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Summary Short-circuited toad bladders were loaded with K42 from the serosal medium in a chamber stirred by rotating impellers. The chambers were washed with nonradioactive Ringer's, and all effluent was collected from the two chambers separately for 30-sec intervals for 30 min and counted. Count rate data were fitted to sums of exponentials and analyzed by the methods of compartmental analysis. There are at least two potassium compartments, with half times of 2.42 and 18.48 min. These compartments contain 2.01 and 7.93 Equiv×100 mg dry weight–1, respectively, amounting to 45% of total tissue K. Determinations of the rate of buildup of tracer in the tissue after immersing the bladder in K42 Ringer's confirmed the fact that only a portion of tissue K exchanges even after one hr; thus the rest must have a considerably slower exchange rate. Fluxes at the inside border are far greater than at the outside, as predicted from electrophysiological data. Of the two tissue compartments, only the smaller and faster one appears to be related to Na transport, since only this compartment shows changes after Na removal (unidirectional serosal K fluxes decrease by some 50%) or after the addition of vasopressin (serosal fluxes and pool size increase by over two-fold). The results also are consistent with the operation of a 11 Na–K exchange pump at the serosal border. 相似文献
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Insulin-stimulated sodium transport in toad urinary bladder 总被引:1,自引:0,他引:1
Mammalian and teleost insulins increase active sodium transport by the toad urinary bladder at subnanomolar concentrations. This stimulation is evident within 15 min and persists for hours. Porcine proinsulin and a cross-linked derivative of bovine insulin are less effective than porcine insulin in stimulating the short-circuit current (SCC), indicating the specificity appropriate for activation of sodium transport through an insulin receptor. The initial stimulation by insulin of the SCC is not blocked by pretreatment with actinomycin D, puromycin, cycloheximide, or tunicamycin. However, in the presence of any one of these inhibitors the sustained increase in SCC is blocked and the rise is short-lived, lasting only 45 to 90 min. In amphotericin-treated bladders, the addition of insulin did not further stimulate SCC. 相似文献
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Evaluation of granule exocytosis in toad urinary bladder 总被引:1,自引:0,他引:1
Wade J. B.; McCusker C.; Coleman R. A. 《American journal of physiology. Cell physiology》1986,251(3):C380
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Summary The uptake of C14-urea into everted and noneverted bladder sacs was compared, over short time periods (up to 2 min), with the transepithelial urea fluxes. This method allowed the study of the time course of urea uptake and distribution, while previously this problem was only studied in steady-state conditions. When mucosal uptake was studied no accumulation of C14-urea inside the tissue was observed, indicating that the mucosal border could be the limiting step. Comparative studies of urea and inulin uptake from the serosal side showed that urea equilibrated with the water epithelial cells in less than 30 sec. This accumulation suggested again that the mucosal border is an effective barrier for urea translocation. The kinetics of the increase in urea permeability induced by antidiuretic hormone was also studied and it was similar (T1/2:4.3 min) to the kinetics of the increase in water permeability induced by the hormone (T1/2:5.6 min). A strong parallelism was also observed between the time course of the increases in water and urea permeabilities induced by medium hypertonicity (T1/2 25 and 26 min, respectively). The values obtained for the permeability coefficientk
trans), either at rest or under ADH were similar to those previously reported employing steady-state techniques (28±8 and 432±25 cm·sec–1·10–7, respectively). 相似文献
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《Cell biology international reports》1981,5(3):275-285
Evidence from thin sections and freeze-fracture is presented showing that stretch induces a burst of exocytosis in granular cells of the toad urinary bladder epithelium. Since the role of granule exocytosis in hormonally-induced permeability changes in this tissue has not yet been clarified, we propose that the stretch factor is an important parameter to consider and standardise in future physiological and morpho-functional studies using this model transporting system. 相似文献
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Summary Urea and other small amides cross the toad urinary bladder by a vasopressinsensitive pathway which is independent of somotic water flow. Amide transport has characteristics of facilitated transport: saturation, mutual inhibition between amides, and selective depression by agents such as phloretin. The present studies were designed to distinguish among several types of transport including (1) movement thought a fixed selective membrane channel and (2) movement via a mobile carrier. The former wold be characterized by co-transport (acceleration of labele amide flow in the direction of net flow in the opposite direction). Mucosal to serosal (MS) and serosal to mucosal (SM) permeabilities of labeled amides were determined in paired bladers. Unlabeled methylurea, a particularly potent inhibitor of amide movement, was added to either the M or S bath, while osmotic water flow was eliminated by addition of ethylene glycol to the opposite bat. Co-transport of labeled methylurea and, to a lesser degree, acetamide and urea with unlabeled methylurea was observed. Co-transport of the nonamides ethylene glycol and ethanol could not be demonstrated. Methylurea did not alter water permeability or transmembrane electrical resistance. The demonstration of co-transport is consistent with the presence of ADH-sensitive amide-selective channcels rather than a mobile carrier. 相似文献
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Summary The effect of amiloride on the sensitivity to Na of the mucosal border of toad urinary bladder was investigated by recording Na concentration-dependent transepithelial potential difference (V
t
) and the intracellular potential. When mucosal Na concentration was normal, amiloride added to the mucosal solution at 10–4
m markedly reduced the mucosal membrane potential (V
m
) and altered the potential profile from a two-step type to a well type. Similar changes were observed when Na was totally eliminated from the mucosal medium. The serosal membrane potential was insensitive to amiloride and elimination of mucosal Na. In the absence of amiloride, theV
t
could be described by the Goldman-Hodgkin-Katz equation in the range of mucosal Na concentration from 0 to 16mm, and amiloride extended this concentration range. By using the Goldman-Hodgkin-Katz equation, Na permeability was calculated from the data ofV
t
's obtained in the allowed ranges of Na concentration and compared before and after the addition of amiloride. The results show that Na permeability decreases to 1/600 of control when the maximum dose of amiloride (10–4
m) is applied. The relationship between Na permeability and amiloride concentration is well explained on the basis of assumptions that amiloride binds to the Na site of the mucosal border in one-to-one fashion and in a competitive manner with Na and that Na permeability reduces in proportion to increase in number of the sites bound with amiloride. 相似文献
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Summary The water diffusional permeability, its activation energy and the lipid composition were studied in urinary bladders from toads adapted to different temperatures. It was observed that the unidirectional water flux greatly depends on the temperature at which the experiments are performed. This dependence is greater in the animals adapted to higher temperatures. Toads adapted to cold show strong reduction in the activation energy for water diffusion permeability (from 11.4±1.9 kcal·mol–1 to 4.4±1.1 kcal·mol–1) and an increase of 30% in the amount of total lipids from bladder epithelial cells. There were no significant changes in the phospholipid/cholesterol ratio, composition of the paraffinic chains or protein concentration between toads adapted to both temperatures. The possibility that water translocates through the mucosal border of the toad bladder by partitioning in the polar zone and diffusioning between the hydrocarbon chains of the membrane lipids and that cold adaptation would induce a stronger packing of lipids in the membrane is discussed. 相似文献
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Jorge Flores Patricia Witkum Barbara Beckman Geoffrey W.G. Sharp 《Biochimica et Biophysica Acta (BBA)/General Subjects》1974,362(3):501-508
Studies have been performed on the effect of vasopressin on cyclic AMP content of toad bladders. A prompt increase in cyclic AMP content occurred after exposure to vasopressin, which reached maximal values within 8 min and remained elevated up to 30 min. By a comparison of the dose-response characteristics of vasopressin on cyclic AMP content, with those Na+ transport and osmotic water flow, it was shown that supramaximal concentrations of vasopressin with respect to physiological function generate more cyclic AMP than is required for maximal stimulation of Na+ transport and water flow. Thus, it would seem that a reverse of hormone-sensitive adenylate cyclase is present in this tissue. 相似文献
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Summary The metabolic cost of active sodium transport was determined in toad bladder at different gradients of transepithelial potential, , by continuous and simultaneous measurements of CO2 production and of transepithelial electric current. Amiloride was used to block active sodium transport in order to assess the nontransport-linked, basal, production of CO2 and the passive permeability of the tissue. From these determinations active sodium transport,J
Na, and suprabasal CO2 production,
, were calculated. Since large transients inJ
Na and
frequently accompanied any abrupt change in , steady state conditions were carefully defined.Some 20 to 40 min were required after a change in before steady state of transport activity and of CO2 production were achieved. The metabolic cost of sodium transport proved to be the same whether the bladder expended energy moving sodium against a transepithelial electrical potential grandient of +50 mV or whether sodium was being pulled through the active transport pathway by an electrical gradient of –50 mV. In both cases the value of the ratio
averaged some 20 sodium ions transported per molecule of CO2 produced.When the Na pump was blocked by 10–2
m ouabain, the perturbations of the transepithelial electrical potential did not elicit changes ofJ
Na nor, consequently, of
.The independence of the ratio
from over the range ±50 mV indicates a high degree of coupling between active sodium transport and metabolism. 相似文献
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A. G. Butt C. W. McLaughlin J. M. Bowler R. D. Purves A. D. C. Macknight 《The Journal of membrane biology》1994,142(1):9-20
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.This work was supported by a grant from the Medical Research Council of New Zealand. Purchase of equipment was made possible through grants from the Medical Research Council of New Zealand, the Medical Distribution Committee of the Lottery Board, the University Grants Committee, the Telford Trust, the New Zealand Neurological Foundation and the National Heart Foundation. The expert technical assistance of S. Zellhuber-McMillan is gratefully acknowledged. 相似文献