Regulation of K-Cl cotransport during reticulocyte maturation and erythrocyte aging in normal and sickle erythrocytes.

The age/density-dependent decrease in K-Cl cotransport (KCC), PP1 and PP2A activities in normal and sickle human erythrocytes, and the effect of urea, a known KCC activator, were studied using discontinuous, isotonic gradients. In normal erythrocytes, the densest fraction (d approximately 33.4 g/dl) has only about approximately 5% of the KCC and 4% of the membrane (mb)-PP1 activities of the least-dense fraction (d approximately 24.

Stimulation of human erythrocyte K-Cl cotransport and protein phosphatase type 2A by n-ethylmaleimide: role of intracellular Mg++.

An increase in the activity of membrane-associated protein phosphatase type 1 (mb-PP1) is associated with stimulation of erythrocyte K-Cl cotransport (KCC). We have recently proposed that membrane-associated protein phosphatase type 2A (mb-PP2A) is also involved in KCC regulation by cell swelling (Bize et al., 1999.

Serine/threonine protein phosphatases and regulation of K-Cl cotransport in human erythrocytes.

Activation of K-Cl cotransport is associated with activation of membrane-bound serine/threonine protein phosphatases (S/T-PPases). We characterize red blood cell S/T-PPases and K-Cl cotransport activity regarding protein phosphatase inhibitors and response to changes in ionic strength and cell size. Protein phosphatase type 1 (PP1) activity is highly sensitive to calyculin A (CalA) but not to okadaic acid (OA).

Stimulation of membrane serine-threonine phosphatase in erythrocytes by hydrogen peroxide and staurosporine.

Indirect evidence has suggested that K-Cl cotransport in human and sheep erythrocytes is activated physiologically by a serine-threonine phosphatase. It is activated experimentally by H2O2 and by staurosporine, a kinase inhibitor. Activation by H2O2 and staurosporine is inhibited by serine-threonine phosphatase inhibitors, suggesting that the activators stimulate the phosphatase.

H2O2 activates red blood cell K-Cl cotransport via stimulation of a phosphatase.

K-Cl cotransport is involved in volume regulation in a number of cell types. Cell swelling stimulates K-Cl cotransport, probably by inhibition of a volume-sensitive kinase. K-Cl cotransport can also be activated by oxidants and thiol reagents.

Staurosporine, a protein kinase inhibitor, activates K-Cl cotransport in LK sheep erythrocytes.

K-Cl cotransport can participate in volume regulation in a number of cell types. Swelling activation of K-Cl cotransport in sheep erythrocytes proceeds by a two-step process, A<-->B<-->C (Dunham et al., J.

Hydrogen peroxide is involved in hamster sperm capacitation in vitro.

We have investigated the possibility that the generation of hydrogen peroxide (H2O2) by spermatozoa plays a physiological role during capacitation. Capacitation is defined as the incubation period required for fertilization in mammals. Capacitation culminates in an exocytotic event, the acrosome reaction (AR).

Studies of stallion sperm survival: preservation of progressive motility of stallion spermatozoa by low ionic strength media.

Preservation of stallion sperm forward motility was studied using a video recording system in semen diluted with media of different ionic strength and sodium content. After 8 hr of incubation at room temperature, semen diluted in a low ionic strength media containing sucrose displayed 65 +/- 9% motility with 68 +/- 3% of the motile sperm showing forward motility (diameter of head trajectory greater than or equal to 80 microns). In contrast, sperm populations diluted and incubated with a normal ionic strength media containing sodium had 56 +/- 7% motile sperm of which only 36 +/- 7% displayed forward progression.

Na-K-C1 cotransport and Na pump in red cells of young blacks and blood pressure response to salt loading.

The functional operation of Na and K fluxes, mediated by the furosemide-sensitive (FS) Na-K-C1 cotransport and the ouabain-sensitive Na pump, was studied in a representative young (18-22 years) black population. Cation fluxes (mumol/L cell X hour = FU) were studied prior to salt loading; results were compared with the blood pressure response to chronic oral salt loading. All subjects (n = 26) took 10 g/day NaC1 plus their usual diets for 14 days.

Epinephrine decreases the potassium requirements of hamster sperm capacitation: furosemide blocks the effect of epinephrine.

We have investigated the possibility of an interaction between epinephrine and external potassium in the stimulation of the hamster sperm acrosome reaction and in vitro fertilizing ability. We have found that: 1) K+ is required late in capacitation and/or the acrosome reaction, but is not required for penetration of the zona pellucidae, 2) the presence of epinephrine (50 microM) in the capacitation medium lowers the K+ requirements of these processes and 3) furosemide, a cotransport inhibitor, blocks the acrosome reaction, in vitro fertilization, and inhibits the stimulation by epinephrine. These results support the idea that there is an interaction between epinephrine as a stimulus to the secretion of the acrosomal content, and K+ as a mediator of the stimulus signal.

Cotransport of lithium and potassium in human red cells.

This paper reports the presence of human red cells of an additional ouabain-insensitive transport pathway for lithium ions, the Li-K cotransport. Several kinds of observations support this conclusion. Cells loaded to contain only K, Na, or Li do not exhibit furosemide-sensitive efflux.

Ki+-Nao+ countertransport in erythrocytes of patients with essential hypertension.

1. We describe in this paper a new ouabain-insensitive pathway for Na+ and K+ in human erythrocytes. K+ efflux was measured in cells loaded by the p-chloromercuribenzene-sulphonate (PCMBS) procedure to contain approximately equal amounts of Na+ and K+.

Na countertransport and cotransport in human red cells: function, dysfunction, and genes in essential hypertension.

We describe in this paper studies on the modes of operation of ouabain-insensitive sodium transport systems in red cells of normotensive and hypertensive patients. We have extensively investigated the properties of Na countertransport and cotransport in order to clarify whether they are two different proteins or one transport protein with two modes of operation. Several criteria of discrimination between the two pathways are described: They differ in their affinity for Na and Li, sensitivity to several inhibitors, changes in cell volume, and chloride replacement by nitrate.

Superoxide dismutase and superoxide radical in Morris hepatomas.

Total superoxide dismutase (SOD) and manganese superoxide dismutase (Mn SOD) specific activities were measured in tissue homogenates and in isolated mitochondria from normal rat liver and three Morris hepatomas of different growth rates. Total SOD and Mn SOD specific activities were decreased in all tumor homogenates when compared to normal liver; the lowest activity was associated with the fastest growing tumor. These results are consistent with the hypothesis that total Mn SOD specific activity is decreased in all tumors.

Modification of ethanol intoxication by dopamine agonists and antagonists.

The effects of ethanol on locomotor activity and rotarod performance were measured in the rat. Apomorphine (4 mg/kg s.c.

Superoxide dismutase activity of normal murine liver, regenerating liver, and H6 hepatoma.

By means of both direct assay and gel electrophoresis, normal A/J mouse liver was shown to possess both Cu-Zn and Mn superoxide dismutase (SD) activity. H6 hepatoma cells contained Cu-Zn SD activity, but no Mn SD activity was detectable. Isolated mitochondria from normal liver contained both forms of the enzyme, but isolated mitochondria from H6 hepatoma cells contained no SD activity.