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Glucose transport in fish erythrocytes: variable cytochalasin-B-sensitive hexose transport activity in the common eel (Anguilla japonica) and transport deficiency in the paddyfield eel (Monopterus albus) and rainbow trout (Salmo gairdneri).

Erythrocytes from individual common eels (Anguilla japonica Temminck and Schlegel) exhibited widely variable initial rates of cytochalasin-B-sensitive 3-O-methyl-D-glucose (3-OMG) zero-trans influx, in the range of 0-19.5 mmol l-cells-1 h-1 (5 mmol l-1 extracellular concentration at 20 degrees C, 50 animals tested). Storage of cells at 4 degrees C in a glucose-containing medium for up to 72 h had no effect on 3-OMG uptake, and there was no correlation between the sugar permeabilities of erythrocytes from different fish and intracellular ATP levels. Adrenaline and noradrenaline increased cytochalasin-B-sensitive 3-OMG transport activity; half-maximal stimulation occurred at catecholamine concentrations in the region of 1 mumol l-1. This catecholamine-induced stimulation of sugar transport appeared to be independent of the basal cytochalasin-B-sensitive 3-OMG permeability of the cells. Kinetically, catecholamines increased the Vm of transport without changing the apparent Km (approx. 1.4 mmol l-1). Saturable 3-OMG influx was inhibited by phloretin, D-glucose, D-deoxyglucose and D-galactose, but not by D-fructose and L-glucose. Transporter stereoselectivity was confirmed by direct measurements of D- and L-glucose uptake. Erythrocytes from two other fish species, Monopterus albus Richardson (paddyfield eel) and Salmo gairdneri Richardson (rainbow trout), unlike those from the common eel, were uniformly deficient with respect to cytochalasin-B-sensitive 3-OMG and D-glucose transport activity. Catecholamines had no effect on sugar uptake in these species.

3-O-Methylglucose↗

Influence of temperature on hexose transport by round spermatids of rats.

Transport of 3-O-methyl-D-[14C]glucose in vitro by round spermatids was more rapid at 34 than at 37 degrees C and more rapid at 37 than at 40 degrees C. The difference between values at 34 and 37 degrees C was reversed by changing the spermatids from 37 to 34 degrees C at various times during a total incubation of 1 h. The effect of 37 degrees C relative to 34 degrees C was exerted on Km for the transport process; Vmax was unaltered. Inhibition by 37 degrees C relative to 34 degrees C was exerted on influx; efflux was the same at both temperatures. A fraction of cells from rat testis enriched in spermatocytes but free from spermatids showed greater transport at 37 than at 34 degrees C. These results indicate that one metabolic effect of body temperature (37 degrees c) as opposed to scrotal temperature (34 degrees C) is inhibition of glucose transport into round spermatids. This may contribute to the defective spermatogenesis that can occur in abdominal testes.

3-O-Methylglucose↗

Evidence that downregulation of hexose transport limits intracellular glucose in 3T3-L1 fibroblasts.

Measurements of initial glucose entry rate and intracellular glucose concentration in cultured cells are difficult because of rapid transport relative to intracellular volume and a substantial extracellular space from which glucose cannot be completely removed by quick exchanges of medium. In 3T3-L1 cells, we obtained good estimates of initial entry of [14C]methylglucose and D-[14C]glucose with 1) L-[3H]glucose as an extracellular marker together with the [14C]glucose or [14C]methylglucose in the substrate mixture, 2) sampling times as short as 2 s, 3) ice-cold phloretin-containing medium to stop uptake and rinse away the extracellular label, and 4) nonlinear regression of time courses. Methylglucose equilibrated in two phases--the first with a half-time of 1.7 s and the second with a half-time of 23 s; it eventually equilibrated in an intracellular space of 8 microliters/mg protein. Entry of glucose remained almost linear for 10 s, making its transport kinetics easier to study (Km = 5.7 mM, Vmax = 590 nmol.s-1.ml-1 cell water). Steady-state intracellular glucose concentration was 75-90% of extracellular glucose concentration. Cells grown in a high-glucose medium (24 mM) exhibited a 67% reduction of glucose-transport activity and a 50% reduction of steady-state ratio of intracellular glucose to extracellular glucose.

Animals↗

Hexose shunt dehydrogenase activity in leukocytes isolated from bovine milk.

A spectrophotometric method for measuring small changes in light absorption at specific wavelengths from highly turbid suspensions of biological material (dual-beam spectrophotometry) has been used to measure glucose-6-phosphate oxidation and nicotinamide adenine dinucleotide phosphate (oxidized) reduction catalyzed by lysates of milk leukocytes. This reduction was correlated .84 with cellular protein concentration and, therefore, with cell concentration.

Animals↗