Electroneutral uptake and electrogenic secretion of a fluorescent bile salt by rat hepatocyte couplets

1993 ◽  
Vol 264 (2) ◽  
pp. G220-G230 ◽  
Author(s):  
S. A. Weinman ◽  
J. Graf ◽  
C. Veith ◽  
J. L. Boyer
Keyword(s):  
Bile Salt ◽  
Time Course ◽  
Organic Anion ◽  
Hepatic Uptake ◽  
Membrane Voltage ◽  
Electrogenic Transport ◽  
Anion Secretion ◽  
Uptake Process ◽  
Transmembrane Potential Difference

The role of membrane voltage as a driving force for the hepatic uptake and secretion of fluorescent bile salts has been examined in isolated hepatocyte couplets. The present study demonstrates that the fluorescent bile salt derivative (N-[7-(nitrobenz-2-oxa- 1,3-diazol-4-yl)]-7-amino-3 alpha, 12 alpha-dihydroxy-5-cholan-24-oyl)-2-aminoethanesulfonate (7 beta-NBD-NCT) is taken up into hepatocytes by a saturable process with a Kt of 2.7 microM. Uptake rate was reduced by only 22% after total Na+ replacement and was independent of transmembrane potential difference over a range of -135 to +25 mV. In contrast, secretion into the canalicular space was strongly dependent on membrane voltage over the range from -34 to 0 mV in a manner consistent with electrodiffusion of an anion. Fitting the secretion time course to that predicted by electrodiffusion demonstrated that only approximately 50% of total secretion can result from electrodiffusion. Studies in isolated perfused liver confirmed this observation that depolarization caused a decrease in bile salt secretion rate. These results demonstrate that 7 beta-NBD-NCT is transported by a neutral uptake process at the sinusoidal membrane and is secreted across the canalicular membrane in part by electrogenic transport. This suggests that voltage changes could be a common pathway resulting in impaired organic anion secretion in diverse cholestatic syndromes.

scholarly journals Fluorescence analysis of the interaction of isometamidium with Trypanosoma congolense

1993 ◽  
Vol 292 (1) ◽  
pp. 31-35 ◽  
Author(s):  
D Zilberstein ◽  
J Wilkes ◽  
H Hirumi ◽  
A S Peregrine
Keyword(s):  
Time Course ◽  
Fluorescence Analysis ◽  
Trypanosoma Congolense ◽  
Sub Saharan Africa ◽  
Temperature Dependent ◽  
Intact Cells ◽  
Uptake Process ◽  
Sub Saharan

Isometamidium chloride (Samorin) is the only compound recommended for prophylaxis against bovine trypanosomiasis in sub-Saharan Africa. The fluorescence property of this compound was used to investigate the interaction of the molecule with in vitro-derived bloodstream forms of Trypanosoma congolense IL 1180. Incubation of isometamidium with trypanosomes at 37 degrees C for 180 min resulted in a gradual alteration of the lambda max. with time (from 600 to 584 nm) and an increase in the intensity of trypanosome-associated fluorescence of approx. 2-fold. The alteration in fluorescence was temperature-dependent and inhibited by the addition of N-ethylmaleimide. In contrast, with intact cells addition of digitonin caused a rapid increase in fluorescence intensity to approximately four times that observed with intact cells. Uptake of isometamidium was also determined using radiolabelled drug; the results indicated that the time course of the uptake process resembled the fluorescence profile and was temperature-dependent. The results therefore indicate that the alteration of fluorescence is due to interaction of isometamidium with an intracellular component(s) and that isometamidium is transported across the plasma membrane via a protein carrier. The data also indicate that the described fluorescence technique can be used to investigate the role of membrane transport in resistance to isometamidium.

Aluminium Uptake by Excised Roots of Cabbage, Lettuce and Kikuyu Grass

1979 ◽  
Vol 6 (6) ◽  
pp. 643 ◽  
Author(s):  
DO Huett ◽  
RC Menary
Keyword(s):  
Time Course ◽  
Cation Exchange Resin ◽  
Effect Of Temperature ◽  
Metabolic Inhibitor ◽  
Rapid Phase ◽  
Uptake Process ◽  
Accumulation Phase ◽  
Kikuyu Grass ◽  
Two Phases

Uptake of aluminium by excised roots of cabbage, lettuce, kikuyu grass and by a weakly acidic cation exchange resin was studied. The role of metabolism was also investigated, using a range of temperatures and a metabolic inhibitor. The time course of uptake had two phases: an initial rapid phase of about 60 min duration for all species and a slower accumulation phase that was pronounced for cabbage and lettuce and almost absent for kikuyu grass. During the initial phase, calcium was exchanged by aluminium. A small fraction of the aluminium taken up by excised roots could not be desorbed by dilute buffer; this was thought to result from irreversible binding of aluminium directly to exchange sites. Evidence is also presented for a third component for aluminium uptake involving diffusion through the plasmalemma. The non-metabolic nature of the entire uptake process was confirmed by the negligible effect of temperature (1-30°C) and enhancement by 2,4-dinitrophenol on the rate of uptake. Aluminium uptake at pH 4.2 exceeded that at pH 4.0, probably due to the lowering of net charge density of aluminium ions as pH is increased.

Collagen-Induced Platelet Aggregation: -Evidence Against the Essential Role of Platelet Adenosine Diphosphate

1979 ◽  
Vol 42 (04) ◽  
pp. 1193-1206 ◽  
Author(s):  
Barbara Nunn
Keyword(s):  
Platelet Aggregation ◽  
Time Course ◽  
Essential Role ◽  
Atp Release ◽  
Adenosine Diphosphate ◽  
Human Platelets ◽  
High Doses ◽  
Induced Aggregation

SummaryThe hypothesis that platelet ADP is responsible for collagen-induced aggregation has been re-examined. It was found that the concentration of ADP obtaining in human PRP at the onset of aggregation was not sufficient to account for that aggregation. Furthermore, the time-course of collagen-induced release in human PRP was the same as that in sheep PRP where ADP does not cause release. These findings are not consistent with claims that ADP alone perpetuates a collagen-initiated release-aggregation-release sequence. The effects of high doses of collagen, which released 4-5 μM ADP, were not inhibited by 500 pM adenosine, a concentration that greatly reduced the effect of 300 μM ADP. Collagen caused aggregation in ADP-refractory PRP and in platelet suspensions unresponsive to 1 mM ADP. Thus human platelets can aggregate in response to collagen under circumstances in which they cannot respond to ADP. Apyrase inhibited aggregation and ATP release in platelet suspensions but not in human PRP. Evidence is presented that the means currently used to examine the role of ADP in aggregation require investigation.

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