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Adenosine uptake in cultured astrocytes is dependent on various ions and energy metabolism. The Na(+)-gradient plays an important role, since nigericin, ouabain, amiloride and substitution of Na+ with choline inhibited adenosine uptake. The proton-gradient was of importance, since carbonylcyanide m-chlorophenylhydrozone (CCCP) and omeprazole also inhibited adenosine uptake. Furthermore, adenosine uptake was dependent on Cl- anion. Substitution of Cl- with isethionate, as well as DIDS or furosemide inhibited adenosine uptake. Adenosine uptake was also sensitive to Ca2+ gradient, removal of extracellular Ca2+ and calcimycin inhibited adenosine uptake. Adenosine uptake was not dependent on extracellular K+ and was not affected by valinomycin. Although, K(+)-channel openers (BRL 34195 and nicorandil) as well as the K(+)-channel antagonist, glyburide, inhibited adenosine uptake, the inhibitory effect of BRL 34915 was not antagonized by glyburide. Rotenone and 2,4-dinitrophenol also inhibited adenosine uptake. Ionic dependence and metabolic energy dependence of adenosine uptake suggest that uptake is primarily an active process.

Potassium is tightly regulated within the extracellular compartment of the brain. Nonetheless, it can increase 3- to 4-fold during periods of intense seizure activity and 10- to 20-fold under certain pathological conditions such as spreading depression. Within the central nervous system, neurons and astrocytes are both affected by shifts in the extracellular concentration of potassium. Elevated potassium can lead to a redistribution of other ions (e.g., calcium, sodium, chloride, hydrogen, etc.) within the cellular compartment of the brain. Small shifts in the extracellular potassium concentration can markedly affect acid-based homeostasis, energy metabolism, and volume regulation of these two brain cells. Since normal neuronal function is tightly coupled to the ability of the surrounding glial cells to regulate ionic shifts within the brain and since both cell types can be affected by shifts in the extracellular potassium, it is important to characterize their individual response to an elevation of this ion. This review describes the results of side-by-side studies conducted on cortical neurons and astrocytes, which assessed the effect of elevated potassium on their resting membrane potential, intracellular volume, and their intracellular concentration of potassium, sodium, and chloride. The results obtained from these studies suggest that there exists a marked cellular heterogeneity between neurons and astrocytes in their response to an elevation in the extracellular potassium concentration.

In astrocytes, as [K+]o was increased from 1.2 to 10 mM, [K+]i and [Cl-]i were increased, whereas [Na+]i was decreased. As [K+]o was increased from 10 to 60 mM, intracellular concentration of these three ions showed no significant change. When [K+]o was increased from 60 to 122 mM, an increase in [K+]i and [Cl-]i and a decrease in [Na+]i were observed. In neurons, as [K+]o was increased from 1.2 to 2.8 mM, [Na+]i and [Cl-]i were decreased, whereas [K+]i was increased. As [K+]o was increased from 2.8 to 30 mM, [K+]i, [Na+]i and [Cl-]i showed no significant change. When [K+]o was increased from 30 to 122 mM, [K+]i and [Cl-]i were increased, whereas [Na+]i was decreased. In astrocytes, pHi increased when [K+]o was increased. In neurons, there was a biphasic change in pHi. In lower [K+]o (1.2-2.8 mM) pHi decreased as [K+]o increased, whereas in higher [K+]o (2.8-122 mM) pHi was directly related to [K+]o. In both astrocytes and neurons, changes in [K+]o did not affect the extracellular water content, whereas the intracellular water content increased as the [K+]o increased. Transmembrane potential (Em) as measured with Tl-204 was inversely related to [K+]o between 1.2 and 90 mM, a ten-fold increase in [K+]o depolarized the astrocytes by about 56 mV and the neurons about 52 mV. The Em values measured with Tl-204 were close to the potassium equilibrium potential (Ek) except those in neurons at lower [K+]o. However, they were not equal to the chloride equilibrium potential (ECl) at [K+]o lower than 30 mM in both astrocytes and neurons.(ABSTRACT TRUNCATED AT 250 WORDS)

Water and electrolyte contents, cell pH, membrane potential and 125I- uptake were determined in cultured follicular cells of turtle thyroid. The Na+, K+ and Cl- concentrations in the cultured thyroid cells were 59.2, 119.0 and 50.9 mmol/l cell water respectively. Treatment with TSH (10 mu./ml for 24 h) increased the K+ and Cl- and decreased the Na+ concentrations in cells. The water and protein contents of these cells were 81.6 and 8.7 g/100 g cells respectively. The cell pH was 6.91. With glass micro-electrodes, the resting membrane potential of thyroid cells cultured in Medium 199 averaged 33.9 +/- 0.63 mV which is slightly higher than 29.8 +/- 1.6 mV as calculated from the data on the uptakes of [14C]methyltriphenylphosphonium and 3H2O by the cells. The potential varied linearly with the log of external K+ concentration (between 15 and 120 mmol/l) with a slope of about 24 mV per tenfold change in K+ concentration. Both TSH and cyclic AMP depolarized the cell membrane. Calculations based on the values for the electrolyte concentrations in cells and in culture medium indicated that Na+, K+ and Cl- were not distributed according to their electrochemical gradients across the cell membrane. Na+ was actively transported out of the cells and K+ and Cl- into the cells. Follicular cells of turtle thyroid cultured in the medium without addition of TSH formed a monolayer. Their iodide-concentrating ability was low and they did not respond to TSH with an increase in iodide uptake. In contrast, cells cultured in medium containing TSH tended to aggregate and organize to form follicles. They had higher ability to concentrate iodide and respond to TSH.

1. Choroid plexus from the lateral ventricle in the adult rat was found to contain approximately 54 m-equiv Na(+) and 89 m-equiv K(+) per kg wet tissue.2. The total water (79%), the extracellular space (21%) and the red blood cell volume (8-9%) in choroid plexus were quantified separately by analysing the distribution of [(14)C]antipyrine, [(14)C]inulin and (51)Cr-tagged erythrocytes, respectively, between this tissue and plasma water.3. The tissue electrolyte data together with the compartmental (space) data were used to calculate an average concentration of Na(+) (39 m-equiv/kg cell H(2)O) and of K(+) (144) in the choroid cell.4. Under various experimental conditions known to stimulate or inhibit the Na(+)-K(+) transport system in other tissues, there were significant changes (10-40 m-equiv/kg cell H(2)O) in the concentrations of both these cations in the plexus epithelial cells.5. Choroid cell K(+) was not independent of the concentration of K(+) in plasma since substantial fluctuations in cell K(+) occurred in rats rendered either hypo- or hyperkalaemic; also, the choroid cell apparently cannot maintain a constant gradient between itself and c.s.f. in the face of kalaemic disturbances.6. Evidence is offered to support the hypothesis that the choroid plexus of the lateral ventricle has a Na(+)-K(+) pump, the operation of which contributes to the maintenance of K(+) homoeostasis in the C.N.S.

Chloride and potassium activities inside the lumen and the transepithelial potentials of control and drug-treated turtle thyroid follicles were determined simultaneously by ion-selective and conventional 3 M-KCl micro-electrodes, respectively. Water and electrolyte contents of these thyroid tissues were determined chemically after each experiment. Cellular and luminal concentrations of Cl- and K+ in control and drug-treated thyroid glands were derived from the data obtained. Both Cl- and K+ equilibrium potentials across the follicular cell membranes calculated from their concentration gradients are not identical to their corresponding membrane potentials measured directly. Thus, transport of both ions occurs. Thyrotrophin, SITS and ouabain increased both cellular and luminal Cl- concentrations. Acetazolamide increased the cellular but did not alter the concentrations. Furosemide and perchlorate markedly increased the cellular but decreased the luminal Cl- concentrations. There is a discrepancy in the intracellular K+ concentration between values derived from electrometric data as measured by ion-selective micro-electrodes and those calculated from the chemical analyses. The electrometrically determined intracellular K+ concentration is smaller. Intracellular Cl- concentrations derived from measurements with the Cl- ion-selective micro-electrodes were in the same range as those calculated from chemical analyses. Thus, either method provides accurate values for intracellular Cl- concentrations.

Elevation of the extracellular potassium concentration above its "resting" level of 5.4 mM stimulated uptake of 45Ca2+ in primary cultures of astrocytes. This effect was only observed when cells were exposed to excess potassium shortly after their exposure to 45Ca2+ and was potently inhibited (IC50 congruent to 3 nM) by the calcium channel blocker nimodipine. In contrast, nimodipine exerted little effect on unstimulated basal uptake of 45Ca2+. These findings suggest that the therapeutic benefit of calcium channel blockers in epilepsy may result in part from the ability of these drugs to prevent calcium entry into astrocytes during seizures when the extracellular potassium is elevated four- to fivefold above normal.

Cl(-) and K(+) activities in the follicular lumen and the intraluminal potential of the same lumen were measured simultaneously with specific liquid ion-exchanger and 3 M KCl microelectrodes, respectively, in turtle thyroid glands. The Cl(-) equilibrium potential between the thyroid interstitium and the lumen calculated from the measured Cl(-) activities was higher than the directly measured intraluminal potential. These data indicate that Cl(-) is actively transported out of the follicular lumen in turtle thyroid gland. On the other hand, the calculated K(+) equilibrium potential was not different from the directly measured potential, which indicates that K(+) is probably distributed passively according to the electrochemical gradients between the interstitial and the luminal compartments in turtle thyroid gland. Results obtained from the specific liquid ion-exchanger microelectrode studies correlate well with those obtained from previous radioautographic observations and chemical determinations.