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  • 1
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 276 (1978), S. 70-71 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] Together with iron and copper, zinc is one of the trace metals most widely required for biological processes. It is generally considered essential for both plant and animal growth and it is a cofactor of several enzyme systems, such as alcohol dehy-drogenase, carbonic anhydrase and ...
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 358 (1975), S. 135-157 
    ISSN: 1432-2013
    Keywords: Frog Skin Epithelium ; S.C.C. Variations ; Intraepithelial Na Pools-Ouabain-Amiloride
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Experimental conditions are described in which transient and positive current responses across isolated frog skin epithelia can be elicited by sudden addition of Na and Li ions (2–40 mM) in the outer bathing solution. Subsequent return to outer Na (or Li) free conditions produce similar transient current changes but in the opposite direction. Analysis of the curve responses shows that the transient component of each curve is best described by a single, fast exponential term equation in case of Na addition to preparation unpoisonned with ouabain. In contrast, an equation including two exponential terms (a fast and a slow one) is required to fit the curve responses observed across ouabain treated epithelia of if Li is added outside. The transient responses were not significantly altered by substituying Cl for SO4 2− anions. They were completely prevented by Amiloride (5 · 10−5 M), increased by oxytocin (20 mU/ml) and markedly dependent upon the outer Na concentration. Interpreted in term of compartmental analysis, these observations suggest that a) the frog skin epithelium contains 2 separated but communicating compartments having different degrees of accessibility from outside; b) only that compartment filling at a fast rate (0.5 min) is involved in the transepithelial Na transport; c) the other one, filling at a rate of 4 to 7 min, is resplenished only under conditions where the basal pump system has a reduced activity. Tentative localization of these compartment is proposed.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-2013
    Keywords: Adenylate Cyclase ; NaF ; Nephron Segments ; Microenzymatic Technique ; Microdissection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary A method is described, which allows adenylate cyclase activity measurement in single pieces of various nephron segments. Tubular samples of 0.5 to 2 mm length were isolated by microdissection from collagenase treated slices of rabbit kidney. A photograph of each piece was taken in order to measure its length. After a permeabilisation treatment involving preincubation in a hypoosmotic medium and a freezing step, each sample was incubated for 30 min at 30°C in a medium containing high specific [α-32P]-ATP 3·10−4 M, final volume 2.5 μl. The [32P]-cAMP formed was separated from the other labelled nucleotides by filtering the incubate on a dry aluminium oxide microcolumn;3H cAMP was added as a tracer for measuring cAMP recovery. The sensitivity of the method was found to be a few fentomoles (10−15 M) cAMP. cAMP generation increased linearly as a function of the incubation time up to more than 30 min, and as a function of the length of the segment used. Control and fluoride (5 mM) stimulated adenylate cyclase activities were measured in the following segments of the nephron: early proximal convoluted tubule (PCT), pars recta of the proximal tubule (PR), thin descending limb of the loop (TDL), cortical portion of the thick ascending limb (CAL), distal convoluted tubule (DCT), first branched portion of the collecting tubule (BCT), further cortical (CCT) and medullary (MCT) portions of the collecting tubule. Mean control adenylate cyclase activity varied from 7 (PR) to 75 (BCT) fmoles/mm/30 min. Fluoride addition resulted in a 10 (BCT) to 50 (PR) fold increase in enzyme activity. Series of replicates gave a scatter equal to ±20% (S.D. as a per cent of the mean). The method described appears to be suitable to determine which nephron segments contain hormone-dependent adenylate cyclase.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-2013
    Keywords: Micropuncture ; Dog ; Mg, Ca, Na, K, Pi reabsorption ; Electron probe analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Micropuncture and clearance experiments were done on normal dogs to investigate magnesium handling by proximal and distal nephron segments. Tubular fluid electrolytes were analyzed with the electron microprobe. Tubular fluid to ultrafilterable magnesium ratio (TF/UF magnesium) was observed to rise above unity but less than the TF/P inulin ratio generated along the proximal tubule. This is in contrast to the other major cations, the ratios of which remain close to unity as water is abstracted. Tubular fluid obtained from the distal tubule contained less magnesium than the glomerular filtrate (mean TF/UF magnesium of 0.6) indicating the loop of Henle is the major nephron segment reclaiming a significant portion of the filtered load. The faction of filtered load remaining at the distal sampling site was similar to the fraction appearing in the urine (8% vs 7%) indicating very little reabsorption beyond the distal tubule in these normal states.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-2013
    Keywords: Adenylcyclase ; PTH ; Nephron Segments ; Rabbit Kidney
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary PTH sensitive adenylate cyclase activity was measured in 9 different segments of the nephron, isolated by microdissection from collagenase-treated rabbit kidney slices. The enzyme of the following segments was stimulated by PTH, 1 U/ml: PCT. (proximal convoluted tubule); PR (pars recta); CAL (cortical portion of the thick ascending limb); DCT (distal convoluted tubule); BCT (first, branched portion of the collecting tubule); the segments which did not respond to PTH were: TDL (thin descending limb); MAL (medullary portion of the thick ascending limb); CCT (cortical portion of the collecting tubule distally adjacent to BCT); MCT (collecting tubule from the outer medulla). PTH sensitive adenylate cyclase per mm tubule in PR was half that measured in PCT. Half maximal stimulation corresponded to 50–100 mU/ml PTH (1–2×10−8M) in both PCT and PR, and to about 350 mU/ml in CAL. PTH (1 U/ml) stimulation factors ranged from 5 to 60 depending on the structures. It is concluded that in addition to PCT and PR, CAL and BCT might be target structures involved in the physiological actions of PTH on the kidney.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-2013
    Keywords: Arginine Vasopressin ; Adenylate Cyclase ; Single Tubules ; Rabbit Kidney ; Microdissection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary AVP dependent adenylate cyclase activity was measured in single pieces of 8 different tubular segments isolated from collagenase treated rabbit kidneys. High responses were observed in all the tested portions of the collecting tubule, that is its cortical branched part (BCT), its cortical straight part (CCT) and its outer medullary part (MCT). Dose response curves indicated in CCT: 2 fold threshold stimulation at 10−11 M AVP, 27 fold stimulation at 10−6 M AVP, half maximal stimulation at about 10−9 M AVP. Both the medullary (MAL) and, to a lesser exten, the cortical (CAL) portions of the thick ascending limb were also observed to contain AVP sensitive adenylate cyclase (for MAL: 2 fold threshold stimulation at 10−9 M AVP, 9 fold stimulation at 10−7 M AVP, half maximal stimulation at 5×10−9 M AVP). In contrast, nearly no responsiveness to AVP was observed in the proximal convoluted tubule, in the thin descending limb of the loop and in the distal convoluted tubule (DCT). The limited response obtained in DCT (which is a structure generally considered as a target site for AVP) as well as the clearcut effect elicited by AVP in MAL (the functioning of which is not known to be controlled by ADH) were unexpected observations; their possible physiological implications will be discussed.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 358 (1975), S. 159-177 
    ISSN: 1432-2013
    Keywords: Frog Skin Epithelium ; S.C.C. Variations ; Intracellular Sodium ; Amiloride ; Tracers Wash-Out
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The ionic composition of the current crossing each membrane of the frog skin epithelia during a) the positive and transient current responses elicited by sudden addition of Na ions to the outer Na free medium b) the negative and transient current responses observed when Na loaded preparations are suddenly exposed to outer Na free solution was determined using isotopic techniques. It is shown that Na ions carry the current across the outer membrane while K ions are mainly involved in the transfert of charges across the inner membrane. The amount of Na accumulated by the epithelial cells during the responses is correlated to the area under the transient part of the current responses. Determinations of24Na uptake at different time of these transient positive responses show that the unidirectional Na influx across the outer membrane decreases as a function of time. It is suggested that the intracellular Na concentration might control the Na uptake mechanism across the outer membrane. During the negative responses, the Na efflux into the outer medium is highly correlated, either in time course or magnitude, to the current response. Both Na efflux and negative current are sensitive to amiloride, suggesting that the mechanism of Na uptake by the frog skin also is able to promote Na movement out of the epithelial cells.
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  • 8
    ISSN: 1432-2013
    Keywords: Adenylate cyclase ; Catecholamines ; Rabbit kidney ; Single tubules ; Nephron micro-dissection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The sensitivity to catecholamines of the adenylate cyclase (AC) activity contained in single tubule samples was investigated on 10 different well defined segments, isolated by microdissection from collagenase treated rabbit kidneys. No responsiveness to isoproterenol (10−6 M) was observed in the proximal tubule (convoluted and straight portions), the thin descending and thick ascending limbs of the loop of Henle, and the first (“bright”) portion of the distal convoluted tubule (DCTb); in contrast high responses (stimulation factors: 4 to 6 fold) were obtained in the second (“granular”) portion of the distal convoluted tubule (DCTg), as well as in both the “granular” (CCTg) and the “light” (CCTl) portions of the cortical collecting tubule. In absolute value, however, the CCTl response was definitely lower than those measured in DCTg and CCTg, as is its control activity. In the medullary portion of the collecting tubule, the AC response to isoproterenol was rather poor both in absolute and relative terms. Dose-response curves measured on DCTg samples indicated a threshold response with an isoproterenol concentration below 10−8 M; half maximal effect corresponded to about 3×10−8 M. CCTl sensitivity to isoproterenol was of the same order of magnitude. Isoproterenol as well as norepinephrine effects in DCTg and CCTl were completely suppressed by 10−4 M propranolol, indicating that the observed AC stimulation was mediated via receptors of the β type. In β blocked CCTl, 10−6 M norepinephrine did not inhibit vasopressin-induced AC stimulation; in the presence of 10−6 M norepinephrine, 10−4 M phentolamine resulted in no additional AC stimulation in DCTg and CCTl; these data suggest the absence of α receptors inhibiting AC activity in these structures. In DCTg, AC stimulation induced either by 10−6 M isoproterenol or by 1 U/ml PTH were observed to be additive when the two hormones were given together. The presence of catecholamine-dependent AC activity in three distal portions of the rabbit nephron is discussed in relation to its possible physiological implications.
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