Energetics of sugar transport by isolated intestinal epithelial cells: effects of cytochalasin B

1979 ◽  
Vol 237 (1) ◽  
pp. C56-C63 ◽  
Author(s):  
G. A. Kimmich ◽  
J. Randles
Keyword(s):  
Steady State ◽  
Epithelial Cells ◽  
Transport System ◽  
Cytochalasin B ◽  
Intestinal Epithelial Cells ◽  
Sugar Accumulation ◽  
Facilitated Diffusion ◽  
Intestinal Epithelial ◽  
Passive System ◽  
Concentration Gradients

The capability of isolated intestinal epithelial cells to establish concentration gradients of 3-O-methylglucose (3-OMG) by a Na+-dependent transport system is limited by concomitant function of a Na+-independent, facilitated diffusion transport system. Monosaccharides accumulated by the active system are continuously lost via the passive system, which acts to lower steady-state sugar gradients maintained by the cell. Cytochalasin B is a potent inhibitor of the passive system and allows the cells to establish a sugar gradient that is much higher than normal. When extracellular [3-;OMG] is 1 mM, cytochalasin induces sugar accumulation ratios of 30-;fold (+/- phlorizin) in contrast to control ratios of approximately 10-;fold. When [3-;OMG] is 0.1 mM, cytochalasin (0.1 mM) induces 40-;fold accumulation ratios. When changes in extracellular sugar concentration are considered, steady-state concentration gradients observed are 70-;fold. For a Na:sugar coupling stoichiometry of 1:1, gradients of this magnitude represent the approximate theoretical maximum for a transport system driven exclusively by the transmembrane electrochemical potential for Na+.

alpha-Methylglucoside satisfies only Na+-dependent transport system of intestinal epithelium

1981 ◽  
Vol 241 (5) ◽  
pp. C227-C232 ◽  
Author(s):  
G. A. Kimmich ◽  
J. Randles
Keyword(s):  
Steady State ◽  
Epithelial Cells ◽  
Transport System ◽  
Intestinal Epithelium ◽  
Cytochalasin B ◽  
Intestinal Epithelial Cells ◽  
Intestinal Epithelial ◽  
Entry Rate ◽  
Dependent Transport

The unidirectional influx of alpha-methylglucoside (alpha-MG) by isolated chicken intestinal epithelial cells is 98% inhibited by phlorizin. The remaining 2% of the total influx occurs in the absence of Na+, is not sensitive to phloretin, and is equal to the diffusional entry rate for 2-deoxyglucose. The glucoside is much more strongly accumulated (75-fold) than 3-O-methylglucose (3-OMG) (10-fold). Inhibitors of the serosal sugar carrier (phloretin, cytochalasin B, theophylline, and flavanoids) do not enhance alpha-MG accumulation. It is concluded that the glycoside is not a substrate for the intestinal serosal transport system. Steady-state gradients of the sugar can be represented accurately by a concentrative, phlorizin-sensitive system that is opposed by a diffusional efflux process.

Effects of phloretin and theophylline on 3-O-methylglucose transport by intestinal epithelial cells

1978 ◽  
Vol 234 (3) ◽  
pp. C64-C72 ◽  
Author(s):  
J. Randles ◽  
G. A. Kimmich
Keyword(s):  
Epithelial Cells ◽  
Transport System ◽  
Intestinal Epithelial Cells ◽  
Sugar Transport ◽  
Inhibitory Effect ◽  
Metabolic Effects ◽  
Transport Systems ◽  
Facilitated Diffusion ◽  
Intestinal Epithelial ◽  
Gradient Enhancement

Phloretin and theophylline each exert an immediate inhibitory effect on the Na+-independent, facilitated-diffusion transport system for sugar associated with intestinal epithelial cells. Phloretin inhibits approximately 50% more of the total Na+-independent sugar flux than theophylline. Neither agent has an immediate effect on the Na+-dependent, concentrative sugar transport system, although preincubation of the cells with phloretin causes a significant inhibition. The slowly developing effect is correlated with a decrease in cellular adenosine triphosphate (ATP) and an elevation of intracellular Na+. Other agents which elevate cell Na+ also inhibit Na+-dependent sugar influx, even if ATP levels are not depleted. On the other hand, if ATP is depleted by phloretin under conditions in which the cells do not gain Na+, the inhibitory effect on Na+-dependent sugar flux tends to disappear. The slow-onset phloretin effects are due to transinhibition of the Na+-dependent sugar carrier by cellular Na+. When the passive sugar carrier is inhibited by phloretin or theophylline, the concentrative system can establish an enhanced sugar gradient. Because of the secondary metabolic effects of phloretin, theophylline induces a greater gradient enhancement despite its more limited effect on the passive sugar-transport system. Sugar gradients as large as 20-fold are induced by theophylline, in contrast to 12-fold gradients observed in the presence of phloretin and approximately 7- to 8-fold for untreated cells. These results are discussed in terms of conceptual questions regarding the energetics of Na+-dependent transport systems.

Energetics of Na+-dependent sugar transport by isolated intestinal cells: evidence for a major role for membrane potentials.

1977 ◽  
Vol 233 (5) ◽  
pp. E357
Author(s):  
G A Kimmich ◽  
C Carter-Su ◽  
J Randles
Keyword(s):  
Steady State ◽  
Cytochalasin B ◽  
Driving Forces ◽  
Intestinal Epithelial Cells ◽  
Sugar Transport ◽  
Intestinal Cells ◽  
Intestinal Epithelial ◽  
Ion Diffusion ◽  
Passive System ◽  
Cell Interior

Intestinal epithelial cells isolated from 6-wk-old chickens maintain the capability for Na+-dependent concentrative accumulation of 3-O-methylglucose (3-OMG). Cells depleted of ATP exhibit a transient accumulation of 3-OMG in response to imposed Na+ gradients ([Na+]o greater than [Na+]i) or when transmembrane ion diffusion potentials (cell interior negative) are established. Phlorizin or lack of extracellular Na+ prevents formation of sugar gradients in every case. A nonconcentrative, non-Na+-dependent sugar transport system is also operative in these cells. The latter system is inhibited to various degrees by phloretin, theophylline, cytochalasin B, and a variety of flavonones and flavones, including apigenin. These agents also act to inhibit efflux of sugar from the cell via this pathway. The concentrative system normally operates against a "leak" of sugar through the nonconcentrative carrier. If the passive system is made inoperative by any of the agents named above, a significant enhancement of steady-state sugar gradients maintained by the cells is observed. With cytochalasin B, gradients as large as 30-fold are established. The energy inherent in cellular Na+ gradients cannot account for sugar gradients of this magnitude unless both chemical electrical driving forces are considered. When the passive leak is maximmally inhibited, more than half of the total energy required must be derived from the membrane potential.

On the mechanism of folate transport in isolated intestinal epithelial cells

1981 ◽  
Vol 240 (2) ◽  
pp. G170-G175 ◽  
Author(s):  
Y. Eilam ◽  
M. Ariel ◽  
M. Jablonska ◽  
N. Grossowicz
Keyword(s):  
Steady State ◽  
Epithelial Cells ◽  
Intestinal Epithelial Cells ◽  
State Level ◽  
Metabolic Inhibitors ◽  
Intestinal Epithelial ◽  
Coupled Transport ◽  
Folate Transport ◽  
Kinetics Of ◽  
Stimulation Of

The mechanism of folic acid (FA) uptake was studied in isolated intestinal epithelial cells prepared from 2- to 6-wk-old chicks. The cells accumulated FA, reaching a level of three- to fivefold that at equilibrium. In the presence of the metabolic inhibitors, NaN3 or KCN, FA was taken up only until equilibration while accumulation of FA was inhibited. Addition of these inhibitors at a steady state of FA accumulation caused a release of intracellular FA. The kinetics of FA uptake were found to be saturable (Km = 3.5 x 10(-5) M), indicating a carrier-mediated mechanism. The steady-state level of FA accumulation was higher as the concentration of NA+ in the medium increased from 0 to 120 mM. This stimulation of FA uptake by Na+ was not due to the stimulation of glucose uptake, because in experiments carried out in the presence of phlorizin, a glucose-transport inhibitor, FA accumulation was not diminished. It is suggested that FA is taken up by a Na+-coupled transport system.

scholarly journals Uptake of Manganese from the Manganese-Lysine Complex in Primary Chicken Intestinal Epithelial Cells

Animals ◽  
2019 ◽  
Vol 9 (8) ◽  
pp. 559
Author(s):  
Shiping Bai ◽  
Keying Zhang ◽  
Xuemei Ding ◽  
Jianping Wang ◽  
Qiufeng Zeng ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Mrna Expression ◽  
Transport System ◽  
Intestinal Epithelial Cells ◽  
Mrna Level ◽  
Intestinal Epithelial ◽  
Divalent Metal Transporter 1 ◽  
Divalent Metal Transporter ◽  
Cationic Amino Acid ◽  
Metal Transporter

Organic manganese (Mn) sources can replace inorganic Mn as dietary Mn supplements in poultry. To compare the uptake of Mn from the Mn-lysine complex (MnLys) and MnSO4, we first established the primary chicken intestinal epithelial cells (IECs) model and used it to determine Mn uptake. The MnLys increased the uptake of Mn compared to MnSO4. The uptake of Mn decreased in the IECs with Fe addition in the medium regardless of the Mn sources. The MnLys decreased the Mn2+ efflux transporter ferroportin 1 (FPN1) mRNA level but did not influence the Mn2+ influx transporter divalent metal transporter 1 (DMT1) mRNA expression when compared to MnSO4. The results above indicated that the increase of Mn accumulation for MnLys at least partly was due to the decrease of Mn efflux by reduced FPN1 expression. The addition of N-ethylmaleimide, an L-lysine transport system y+ inhibitor, decreased the uptake of Mn from MnLys but did not affect the uptake of Mn from MnSO4. The cycloheximide, as an L-lysine transport system b0,+ activator, increased the uptake of Mn from MnLys, whereas they did not influence the uptake of Mn from MnSO4. The MnLys increased the system y+ members cationic amino acid transporter (CAT) 1 and CAT2, and system b0,+ components rBAT and b0,+AT mRNA expression when compared to MnSO4. These results suggested that the uptake of MnLys complex might be transported by CAT1/2 and system b0,+, which was different from the ionized Mn2+ uptake pathway. In conclusion, the uptake of Mn from MnLys complex not only might be uptake through the ionized Mn2+ pathway, but also appeared to be transported through the CAT1/2 and system b0,+ in primary chicken IECs.

Characterization and regulation of adenosine transport in T84 intestinal epithelial cells

1998 ◽  
Vol 274 (2) ◽  
pp. G261-G269 ◽  
Author(s):  
Edward C. Mun ◽  
Kevin J. Tally ◽  
Jeffrey B. Matthews
Keyword(s):  
Epithelial Cells ◽  
Binding Sites ◽  
Intestinal Epithelial Cells ◽  
Metabolic Inhibition ◽  
Nucleoside Transport ◽  
Facilitated Diffusion ◽  
Intestinal Epithelial ◽  
Binding Studies ◽  
Simple Diffusion ◽  
Adenosine Transport

Adenosine release from mucosal sources during inflammation and ischemia activates intestinal epithelial Cl−secretion. Previous data suggest that A2b receptor-mediated Cl− secretory responses may be dampened by epithelial cell nucleoside scavenging. The present study utilizes isotopic flux analysis and nucleoside analog binding assays to directly characterize the nucleoside transport system of cultured T84 human intestinal epithelial cells and to explore whether adenosine transport is regulated by secretory agonists, metabolic inhibition, or phorbol ester. Uptake of adenosine across the apical membrane displayed characteristics of simple diffusion. Kinetic analysis of basolateral uptake revealed a Na+-independent, nitrobenzylthioinosine (NBTI)-sensitive facilitated-diffusion system with low affinity but high capacity for adenosine. NBTI binding studies indicated a single population of high-affinity binding sites basolaterally. Neither forskolin, 5′-( N-ethylcarboxamido)-adenosine, nor metabolic inhibition significantly altered adenosine transport. However, phorbol 12-myristate 13-acetate significantly reduced both adenosine transport and the number of specific NBTI binding sites, suggesting that transporter number may be decreased through activation of protein kinase C. This basolateral facilitated adenosine transporter may serve a conventional function in nucleoside salvage and a novel function as a regulator of adenosine-dependent Cl− secretory responses and hence diarrheal disorders.

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