Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.6.1.3 (ATPase)
65,361 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

This study reports on the interaction between transepithelial Na+ transport and H+ secretory and intracellular pH (pHi) regulating mechanisms in the model 'tight' epithelium of frog skin. We have used 22Na isotope fluxes and fixed end-point titration to measure undirectional Na+ fluxes, net Na absorption (J(net)Na) and proton secretion (J(net)H), and electrophysiological techniques (double-barrelled ion-sensitive microelectrodes and cell membrane current--voltage relations) to determine intracellular activities of Na+, Cl- and H+ and the conductance of apical membranes to Na+ (gNa) and of basolateral membranes to K+ (gK). In dilute mucosal solutions or in the absence of a permeant anion (Cl-) or counter-current (open-circuit conditions) to accompany Na+ uptake, the J(net)Na is electrically coupled to J(net)H via an electrogenic apical H+-ATPase (located in mitochondria-rich cells). Both fluxes proceed via mitochondria-rich cells and are inhibited by blockers of carbonic anhydrase and H+-ATPase and stimulated by aldosterone and acid load. In high NaCl-containing mucosal solutions or in short-circuit conditions, the J(net)Na becomes uncoupled from J(net)H and proceeds mainly via the principal cells in the epithelium, in which pHi is regulated by basolateral Na+/H+ and Cl-/HCO3- exchangers. Under these conditions, J(net)Na, gNa and gK vary directly and in parallel with pHi, when pHi is changed by permeable weak acids or bases. There is also co-variance between gNa and pHi accompanying spontaneous variations in J(net)Na and when Na+ transport is stimulated by aldosterone or inhibited with ouabain. We conclude that the level of intracellular H+, modulated by H+ pump and Na+/H+ and Cl-/HCO3- exchangers provides an intrinsic regulation of epithelial Na+ transport.
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PMID:Epithelial pH and ion transport regulation by proton pumps and exchangers. 246 78

1. We compared enzymatic and functional properties of a turtle bladder cell line to those of turtle bladder epithelial cells. Like the original epithelium, the cell line displays carbonic anhydrase activity and acetazolamide inhibits O2 consumption in isolated cells and acidification by cells grown in monolayers. 2. Staining with acridine orange revealed the presence of cytoplasmic orange red vesicles which could be dissipated by NH4Cl or protonophores indicating that these vesicles represent areas of low pH. 3. Addition of ATP to cells permeabilized by digitonin led to reappearance of the red granules suggesting that acidification of the vesicles is mediated by H+-ATPase.
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PMID:Turtle urinary bladder epithelial cell line. 247 49

Immunostaining for Na+, K+-ATPase, carbonic anhydrase (CA) II, and band 3 anion channel glycoprotein was compared in developing and mature human kidneys and in Wilms' tumors. In fetal kidneys, ATPase first appeared in proximal and distal tubules. At birth an adult pattern was present with abundant enzyme in all segments of the distal tubule and lesser amounts in proximal and collecting tubules. CA II was detected in fetal kidneys first in proximal and then in distal tubules and eventually, as in the adult, throughout the nephron. Band 3 glycoprotein was not detected in fetal kidneys and only weak staining was present in the basolateral plasmalemma of intercalated cells in newborn and infant kidneys. The number of cells reactive for band 3 and the intensity of staining in a given cell increased to near adult levels at about 2 years. This finding may provide a partial explanation for the 'physiological acidosis' characterized by a low systemic pH in newborn and young infants. ATPase was present in basolateral membranes of most epithelial cells in nonanaplastic Wilms' tumors but was absent in the epithelial component of two anaplastic Wilms' tumors. CA II was detected only in a few epithelial cells in four tumors. Neoplastic epithelial cells reactive for CA II also stained for ATPase but not vice versa. Band 3 glycoprotein was not detected in any Wilms' tumor. These findings show that the immunohistochemical assessment of protein involved in electrolyte transport provides a further means for determining the relative level of differentiation of tumor cells of epithelial origin and suggest that these methods may be a valuable aid in determining the prognosis of some carcinomas.
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PMID:Immunohistochemical localization of transport mediators in Wilms' tumor: comparison with fetal and mature human kidney. 247 91

Pancreatic ductal cell secretion has not been well characterized due to the difficulty in obtaining sufficient quantities of purified ductal cells. To determine if the MIA PaCa-2 cell line would provide a useful model for in vitro studies of pancreatic ductal cell secretion, the present study was designed to characterize these cells in greater detail. In this investigation, the human pancreatic undifferentiated cell line, MIA PaCa-2, was compared with PANC-1 cells (a human ductal cell line previously characterized), isolated rat and human ducts, acinar cells, and nonpancreatic cell lines. The results indicate that while the morphology of the MIA PaCa-2 cell line is nonpolarized and generally atypical of either ductal or acinar cells, the cell line has retained certain biochemical similarities to ductal cells. Additional morphological studies indicated (a) the presence of intermediate filaments characteristic of epithelial cells, (b) the absence of zymogen granules, and (c) an apparent basolateral plasma membrane localization of Na+, K+-ATPase. Similar to ductal cells, biochemical analyses indicated (a) the presence of Na+, K+-ATPase based on [3H]-ouabain binding assays, (b) high levels of carbonic anhydrase, (c) low levels of gamma-glutamyl transpeptidase, (d) nondetectable levels of amylase, and (e) protein composition and protein synthetic patterns comparable to PANC-1 cells. Finally, as with PANC-1 cells and isolated rat and human ducts, the major sulfated secretory product of MIA PaCa-2 cells was a protein with a molecular weight of approximately 660,000 to 1 million.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Comparative analysis of a human pancreatic undifferentiated cell line (MIA PaCa-2) to acinar and ductal cells. 247 96

1. The effects of ouabain and acetazolamide on the secretion of pancreatic juice stimulated by secretin in anaesthetized dogs were investigated. 2. Intra-arterial injection of ouabain (1-10 micrograms) and acetazolamide (1-10 mg) caused dose-dependent decreases in the volume of pancreatic juice. When both drugs were added together, the inhibitory effects were significantly higher than for each drug alone. 3. The bicarbonate concentration in the pancreatic juice was decreased and the chloride concentration was increased by ouabain and acetazolamide, but sodium and protein concentrations were not modified. 4. The results suggest that the Na+,K+-ATPase and carbonic anhydrase activities play important roles in water and electrolyte secretion, and that ouabain and acetazolamide inhibit secretin-stimulated pancreatic secretion by acting on different systems in the exocrine cells in dogs.
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PMID:Inhibitory effect of ouabain and acetazolamide on secretin-stimulated pancreatic exocrine secretion in anaesthetized dog. 249 20

To examine the oxygen requirement of carbonic anhydrase-dependent sodium reabsorption in the proximal tubule, 18 anaesthetized dogs were studied under conditions of saturated distal NaCl reabsorption; the latter was accomplished by volume expansion (all groups) combined with infusion of loop diuretics (groups 1 and 3). Acetazolamide reduced HCO3- reabsorption by 602 +/- 32 mumol min-1 (55%, group 1) and by 777 +/- 103 mumol min-1 (66%, group 2). This was accompanied with a reduction in sodium reabsorption and oxygen consumption in a molar delta Na/delta O2 ratio of about 45 in both groups of dogs. The delta HCO3/delta O2 ratio averaged 16 +/- 1, which was not significantly different from the theoretical value of 18 expected for transcellular sodium transport by Na+, K+-ATPase. Mannitol (group 3) reduced NaCl reabsorption by 37 +/- 2% without affecting NaHCO3 reabsorption or oxygen consumption significantly. We conclude that carbonic anhydrase-dependent NaCl reabsorption in the proximal tubules is passive, and that NaHCO3 reabsorption is the only important active sodium transport which is sensitive to inhibition of carbonic anhydrase.
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PMID:Low oxygen cost of carbonic anhydrase-dependent sodium reabsorption in the dog kidney. 251 51

The main purpose of this study is to elucidate the effect of adrenocorticoids on Mg2+-HCO3(-)-ATPase and carbonic anhydrase which are thought to be related to anion transport in mammalian intestinal mucosa and renal tubulus. Rat duodenal mucosa, large intestinal mucosa and kidney cortex were excised and homogenized with mannitol-Tris buffer (pH 7.1) and brush border fraction and cytosol were obtained by a differential fractionation procedure. Brush border Mg2+-HCO3(-)-ATPase and cytosol carbonic anhydrase activities in the duodenal mucosa decreased to 70% and 37% of normal values, respectively 5-11 days after adrenalectomy. Adrenalectomy also decreased significantly both enzyme activities in large intestinal mucosa; on the other hand, renal enzyme activities did not change. Four hours after a single injection of 20-80 micrograms/kg of aldosterone, ip, to adrenalectomized rats, Mg2+-HCO3(-)-ATPase and carbonic anhydrase activities in duodenal mucosa increased gradually to normal or near normal in dose-dependent fashion. Both enzyme activities in large intestinal mucosa were also increased by a larger dose of aldosterone. Again, renal enzyme activities were not affected by any dose of aldosterone. In contrast, corticosterone (1 mg and 4 mg/kg) and dexamethasone (50 micrograms 200 micrograms/kg) had no replacement effect on enzyme activities in all organs. These results showed that the mineralocorticoid, but not glucocorticoids, is a regulator of the enzyme activity of Mg2+-HCO3(-)-ATPase and carbonic anhydrase from intestinal mucosa. The true mechanisms by which both enzymes are activated by aldosterone are not clear at present.
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PMID:Further studies on the effect of aldosterone on Mg2+-HCO3(-)-ATPase and carbonic anhydrase from rat intestinal mucosa. 252 25

Heterogeneity in Madin-Darby canine kidney (MDCK) epithelial cells has been reported, however, its details have not been well described. In the present study, we show that subclones obtained from a MDCK cell line could be divided into two morphologically and biochemically distinct cell types with different hormonal responsiveness. Clones of the first type, motile clones, which had extended and flattened cytoplasm, were devoid of carbonic anhydrase activity. Clones of the second type, nonmotile clones, formed colonies of cuboidal cells and showed carbonic anhydrase activity. Motile clones synthesized cAMP in response to arginine vasopressin, prostaglandin E1, and isoproterenol but not glucagon. In contrast, nonmotile clones responded to all of these hormones. These findings suggest MDCK cells have multiple cellular origins. The motile clones have characteristics similar to the principal cells of the collecting system, whereas the nonmotile clones may be derived from the thick ascending limb or the intercalated cell. Our studies also demonstrate a significant influence of culture condition on MDCK cellular behavior (carbonic anhydrase activity, Na+/K+-ATPase activity and vasopressin responsiveness). Therefore, physiologic and biochemical experiments with MDCK cells must be interpreted with reservations about cellular heterogeneity as well as differences induced by culture conditions.
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PMID:Characterization of subclones of Madin-Darby canine kidney renal epithelial cell line. 255 8

The ciliary epithelium of spiny dogfish eyes has previously been used for studies of epithelial electrolyte transport and the results are found to be comparable to those obtained in mammals. In this study we report the ultrastructure and enzyme histochemistry of the ciliary epithelium of spiny dogfish in comparison with that of mammals. In contrast to the mammals, the lens in spiny dogfish is connected to the anterior and middle part of the ciliary epithelium by a broad zonula-like suspensory ligament and by a short ligament to the ventral papilla, while the posterior region is mainly free of zonula-like fibers. The non-pigmented epithelium (NPE) of the posterior region shows numerous membrane infoldings, interdigitations and many mitochondria in the cytoplasm as well as histochemical staining for Na+ K(+)-ATPase and carbonic anhydrase (CA). These findings are similar to those seen in the anterior pars plicata of mammals which is mainly involved in aqueous secretion. In the anterior and middle part, the NPE cells have only a few membrane infoldings and few mitochondria in the cytoplasm, but abundant surfaces of rough endoplasmic reticulum, numerous ribosomes and Golgi material, indicating protein synthesis. Histochemically the cells stain for Na+, K(+)-ATPase but not for CA. These findings are comparable to the pars plana in mammals. In contrast to mammals, where the pigmented epithelium (PE) shows nearly no staining for Na+, K(+)-ATPase but an intensive one for CA, the PE cells of spiny dogfish are heavily stained for Na+, K(+)-ATPase in all regions of the eye, but show nearly no staining for CA.
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PMID:Regional differences in the morphology and enzyme distribution of the spiny dogfish (Squalus acanthias) ciliary epithelium. 255 4

In the proximal tubules, fractional reabsorption remains essentially unchanged during variations in glomerular filtration rate (GFR). Glomerulotubular balance (GTB), defined as the linear relationship between proximal tubular reabsorption and GFR, is quantitatively the most important regulator of tubular reabsorption, which may be stopped by inhibiting Na, K-ATPase activity completely. However, ouabain in doses inhibiting 80% of the Na, K-ATPases, exerts no effect on proximal reabsorption of water, NaCl and NaHCO3. At constant plasma pH, the same relationship between filtered and reabsorbed bicarbonate is obtained whether bicarbonate reabsorption is altered by varying GFR or plasma concentration of bicarbonate. In contrast, a selective rise in plasma NaCl concentration at constant plasma pH (hypernatremia) reduces NaHCO3 reabsorption and fails to stimulate NaCl reabsorption. Other characteristics of proximal tubular reabsorption are that nonreabsorbable solutes, such as mannitol, inhibit water and NaCl reabsorption with little or no change in NaHCO3 reabsorption and renal oxygen consumption. Mannitol reduces the slope of the GTB curve for NaCl but not for NaHCO3. Hypertonic NaHCO3 exerts an osmotic effect on proximal water and NaCl reabsorption comparable to that of mannitol, whereas hypertonic NaCl is without osmotic effect. By reducing plasma pH (hypercapnia at high plasma bicarbonate concentration), the slope of the GTB curves for NaCl and NaHCO3 can be greatly increased. By raising plasma pH either by hypocapnia or bicarbonate loading, proximal reabsorption of NaHCO3 and NaCl is greatly depressed and remains almost unaltered during variations of GFR (abolished GTB). Similarly, carbonic anhydrase inhibitors, such as acetazolamide, reduce the reabsorption of NaCl and NaHCO3 in the same proportion as a rise in plasma pH, and abolish GTB. Examinations of proximal tubular oxygen consumption indicate that the energy requirement for NaHCO3 reabsorption is as expected for transcellular transport by Na, K-ATPases, whereas proximal NaCl reabsorption requires no additional energy. These data indicate that transcellular energy-requiring NaHCO3 reabsorption provides the main osmotic force across the tight junction for paracellular reabsorption of proximal tubular fluid containing NaCl and other solutes of low reflection coefficient. The main factors influencing GTB are the filtered load of bicarbonate, plasma pH and nonreabsorbable solutes in the proximal tubular fluid.
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PMID:Essentials of glomerulotubular balance. 267 97


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