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H Pasantes-Morales

Publications and source records attributed to H Pasantes-Morales.

At least 55 records · Page 3Linked to original sources
PubMed1995

Taurine content in breast milk of Mexican women from urban and rural areas.

The concentration of the most abundant free amino acids in breast milk, taurine, glutamic acid, glycine + threonine, alanine and glycine was measured in breast milk samples obtained from Mexican women from an urban population (38 samples) and from a rural population (106 samples). Free amino acid levels including taurine in the urban group were essentially similar to those reported in samples from American and Canadian women (taurine concentration: 332-357 nmol/ml) but significantly lower content of taurine (237-259 nmol/ml) was found in the rural group. The only other significant difference between the two groups was a 32% higher concentration of alanine in the rural group as compared to the urban group. The observed differences may result from a lower dietary intake of taurine containing food in the rural group as this group reported a restricted consumption of meat, the most abundant dietary source of taurine. Because taurine synthesis is low in primates including human, a decreased external supply of taurine may explain the observed reduction in the taurine content of milk. The increase in alanine may represent a compensatory mechanism for the decrease in taurine.

Amino Acids↗
PubMed1994

Sulfhydryl groups essential for the volume-sensitive release of taurine from astrocytes.

The volume-sensitive [3H]taurine efflux from cultured astrocytes was found to be strongly inhibited by the sulfhydryl group-modifying reagent N-ethylmaleimide (NEM). This maleimide inhibited taurine efflux evoked by 50% hyposmotic solutions with an IC50 of 54 microM. The inhibition by NEM followed pseudo-first order reaction kinetics. A double log plot of the pseudo-first order constant against NEM concentration gave a linear relationship with a slope of 1.2. The data are consistent with a simple bimolecular reaction mechanism in which one molecule of NEM reacts with one sulfhydryl group per transport unit. The membrane-impermeant maleimide derivative 5-eosin maleimide did not affect the volume-stimulated taurine efflux. The sulfhydryl-modifying mercurial reagents mersalyl and p-chloro mercuribenzenesulfonate (0.5-1 mM) increased [3H]taurine efflux under isosmotic conditions and concomitantly decreased the hyposmolarity-evoked efflux. The results demonstrate an essential requirement for sulfhydryl groups for the volume-sensitive taurine efflux.

Animals↗
PubMed1994

Properties of osmolyte fluxes activated during regulatory volume decrease in cultured cerebellar granule neurons.

Efflux pathways for amino acids, K, and Cl activated during regulatory volume decrease (RVD) were characterized in cultured cerebellar granule neurons exposed to hyposmotic conditions. Results of this study favor diffusion pores (presumably channels) over energy-dependent transporters as the mechanisms responsible for the efflux of these osmolytes. The selectivity of osmolyte pathways activated by RVD was assessed by increasing the extracellular concentrations of cations, anions, and amino acids to such an extent that upon opening of the pathway, a permeable compound will enter the cell and block RVD by reducing the efflux of water carried by the exit of intracellular osmolytes. The cationic pathway was found selective for K (and Rb), whereas the anionic pathway was rather unselective being permeable to Cl, nitrate, iodine, benzoate, thiocyanate, and sulfate but impermeable to gluconate. Glutamate and aspartate as K but not as Na salts were permeable through the anion channel. RVD was slightly inhibited by quinidine but otherwise was insensitive to known K channel blockers. RVD was inhibited by 4,4'-diisothiocyanostilbene-2-2'-disulfonic acid (DIDS), niflumic acid, and dipyridamole. Gramicidin did not affect cell volume in isosmotic conditions but greatly accelerated RVD, suggesting that cell permeability to Cl is low in isosmotic conditions but increases markedly during RVD making K permeability the rate limit of the process. The permeability pathway for amino acids activated during RVD as permeable to short chain alpha- and beta-amino acids, but excluded glutamine and basic amino acids.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1994

Impaired cell volume regulation in taurine deficient cultured astrocytes.

Taurine concentration was reduced by 40 and 65%, respectively in rat cerebellar astrocytes grown in a chemically defined medium or in culture medium containing a blocker of taurine transport (GES). Cell volume in these taurine deficient cells was 10%-16% higher than in controls. When challenged by hyposmotic conditions, astrocytes release taurine and this efflux contributes to the volume regulatory decrease observed in these cells. Taurine deficient astrocytes showed a less efficient volume recovery as compared to controls with normal taurine levels. Exposed to 50% hyposmotic medium, astrocytes with normal taurine concentration recovered 60% of their original volume whereas taurine deficient cells recovered only 30-35%. Similarly, in 30% hyposmotic medium, taurine deficient astrocytes recovered only 40% as compared to 75% in controls. No compensatory increases in the efflux of other osmolytes (free amino acids or potassium) were observed during regulatory volume decrease in taurine deficient astrocytes.

Amino Acids↗
PubMed1994

Regulatory volume decrease in cultured astrocytes. I. Potassium- and chloride-activated permeability.

Regulatory volume decrease (RVD) in detached cerebellar astrocytes in culture after acute exposure to hyposmolarity was characterized in this and the accompanying paper [H. Pasantes-Morales, R. A. Murray, R. Sanches-Olea, and J. Moran. Am. J. Physiol. 266 (Cell Physiol. 35): C172-C178, 1994]. RVD was independent of extracellular calcium, was accelerated at pH 8-9 and retarded at pH 6, and was reduced at temperatures < 18 degrees C. The cationic pathway activated by hyposmolarity was specific for K+ and Rb+, since RVD was abolished and secondary swelling occurred when these ions replaced Na+. However, Li+, choline, tris(hydroxymethyl)aminomethane, and glucosamine, all as Cl- salts, did not affect RVD. The anion pathway was unselective, since RVD was inhibited when NaCl was replaced by anion K+ salts with a permeability rank of SCN- = I- > NO3- > Cl- > benzoate > acetate >> SO3- > gluconate. RVD was unaffected by bumetanide (50 microM) and weakly inhibited by furosemide (2 mM). Quinidine but not other K+ channel blockers inhibited RVD, and its effect was reversed by gramicidin. RVD was inhibited by 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid and dipyridamole but not by diphenylamine-2-carboxylate or anthracene-9-carboxylate. These results suggest that diffusion possibly via channels rather than cotransporters is involved in the swelling-activated K+ and Cl- fluxes. Gramicidin did not change astrocyte volume in isosmotic conditions, but greatly accelerated RVD, suggesting that low Cl- permeability in isosmotic conditions markedly increases by swelling, thus making K+ permeability the rate-limiting step for RVD.

Animals↗
PubMed1994

Regulatory volume decrease in cultured astrocytes. II. Permeability pathway to amino acids and polyols.

The permeability of the hyposmolarity-activated pathway to amino acids and polyols in cultured astrocytes was examined following the change in rate and direction of regulatory volume decrease (RVD) when the extracellular concentration of the osmolytes was increased to reverse their intracellular-extracellular concentration gradient. Activation of the pathway by swelling would allow those permeable osmolytes to enter the cell and inhibit RVD. The pathway was found to be permeable to neutral amino acids, with beta-amino acids (beta-alanine = taurine > gamma-aminobutyric acid) more permeable than alpha-amino acids. Glycine, alanine, threonine, phenylalanine, and asparagine, but not glutamine, were permeable through this pathway. Aspartate was more permeable than glutamate, and K+ and not Na+ must be the accompanying cation. Basic amino acids were excluded. The dimension of the amino acid pore activated by hyposmolarity seems to be at the limit of glutamate-glutamine size. Influx rather than efflux of amino acids was observed when extracellular concentration was greater than intracellular concentration, with differences in the amount accumulated by cells correlating with their efficiency as RVD blockers. Influx of taurine (as representative of permeable amino acids) was inhibited by the Cl- channel blockers/exchangers 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (40%) and dipyridamole (85%) , and it is suggested that amino acids permeate through an anion channel. Sorbitol and mannitol, but not inositol, exhibited a small inhibitory effect on the later phase of RVD, whereas inositol slightly accelerated RVD.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1993

Inhibition of volume regulation and efflux of osmoregulatory amino acids by blockers of Cl- transport in cultured astrocytes.

Regulatory volume decrease (RVD) in astrocytes was inhibited by the Cl- exchanger blockers 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), dipyridamole and niflumic acid but not by the Cl- channel inhibitors diphenylamine-2-carboxylate (DPC) and anthracene-9-carboxylate (9-AC). The volume activated efflux of [3H]taurine and [3H]D-aspartate (as marker for glutamate) was similarly affected by these compounds. However, neither RVD nor osmolyte fluxes were significantly reduced by removal of external Cl-, suggesting that an anion exchanger activity is not required for the volume regulatory process. Alternatively, these results suggest that the anion exchanger molecule may function as an unidirectional Cl- channel possibly permeable also to amino acids.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1993

Cell volume regulation in cultured cerebellar granule neurons.

Cultured rat cerebellar granule neurons exposed to solutions of reduced osmolarity, responded initially by swelling followed by a regulatory volume decrease (RVD) which is completed within 15 min. Increasing external osmolarity lead to cell shrinking but no evidence of volume regulation was observed within 1 hr. Replacing Na+ by choline did not affect RVD whereas N-methyl-D-glucamine accelerated the volume recovery and K+ suppressed it completely. The blockade of RVD in high extracellular K+ was only observed when chloride and nitrate but not sulfate or gluconate were the accompanying anions. Replacing intracellular Cl-, by long incubations with gluconate, markedly inhibited RVD. Removal of extracellular Ca2+ or addition of dantrolene which blocks Ca2+ released from intracellular stores had no effect on RVD. Increasing extracellular taurine prevented RVD. These results indicate that membrane permeability to K+, Cl-, and taurine is increased by hyposmolarity and suggest the involvement of these molecules in RVD in granule neurons.

Animals↗
PubMed1993

Characterization of stretch-activated ion channels in cultured astrocytes.

The presence of a stretch-activated channel in rat cerebellar astrocytes in culture is described. This stretch-sensitive channel is K(+)-selective and its open probability increases with suction following a Boltzmann-like distribution with half activation at 45 mm Hg. Kinetic analysis of the single-channel data indicated that there are two open and two closed states and that the shortest time constants of both open and closed states are the most sensitive to suction. A symmetrical two-barrier-one-site permeation model can quantitatively describe the inward rectification of the single-channel current to voltage relations. It is suggested that this stretch-activated channel plays a role in the regulatory volume response of astrocytes to hyposmotic conditions.

Animals↗
PubMed1993

Contribution of organic and inorganic osmolytes to volume regulation in rat brain cells in culture.

In this work we examined the time course and the amount released, by hyposmolarity, for the most abundant free amino acids (FAA) in rat brain cortex astrocytes and neurons in culture. The aim was to evaluate their contribution to the process of cell volume regulation. Taurine, glutamate, and D-aspartate in the two types of cells, beta-alanine in astrocytes and GABA in neurons were promptly released by hyposmolarity, reaching a maximum within 1-2 min. after an osmolarity change. A substantial amount of the intracellular pool of these amino acids was mobilized in response to hyposmolarity. The amount released in media with osmolarity reduced from 300 mOsm to 150 mOsm or 210 mOsm, represented 50%-65% and 13%-31%, respectively, of the total amino acid content in cells. In both astrocytes and neurons, the efflux of glutamine and alanine was higher under isosmotic conditions and increased only marginally during hyposmotic conditions. 86Rb+, used as tracer for K+, was released from astrocytes, 30% and 11%, respectively, in hyposmotic media of 150 mOsm or 210 mOsm but was not transported in neurons. From these results it was calculated that FAA contribute 54% and inorganic ions 46% to the process of volume regulation in astrocytes exposed to a 150 mOsm hyposmotic medium. This contribution was 55% for FAA and 45% for K+ and Cl- in cells exposed to 210 mOsm hyposmotic solutions. These results indicate that the contribution of FAA to the process of cell volume regulation is higher in astrocytes than in other cell types including renal and blood cells.

Amino Acids↗
PubMed1993

Neurons respond to hyposmotic conditions by an increase in intracellular free calcium.

The effect of hyposmotic conditions on the concentration of intracellular free calcium ([Ca2+]i) was studied in cultured cerebellar granule cells and cerebral cortical neurons after loading of the cells with the fluorescent Ca2+ chelator Fluo-3. It was found that in both types of neurons exposure to media with a decrease in osmolarity of 20 to 50% of the osmolarity in the isosmotic medium (320 mOsm) led to a dose dependent increase in [Ca2+]i with a time course showing the highest value at the earliest measured time point, i.e. 40 s after exposure to the hyposmotic media and a subsequent decline towards the basal level during the following 320 s. The response in the cortical neurons was larger than in the granule cells but both types of neurons exhibited a similar increase in [Ca2+]i after exposure to 50 mM K+ which was of the same magnitude as the increase in [Ca2+]i observed in the cortical neurons exposed for 40 s to a medium with a 50% reduction in osmolarity. In both types of neurons the blocker of voltage gated Ca2+ channels verapamil had no effect on the hyposmolarity induced increase in [Ca2+]i. On the contrary, this increase in [Ca2+]i was dependent upon external calcium and could be inhibited partly or completely by the inorganic blockers of Ca2+ channels Mg2+ and La3+. Dantrolene which prevents release of Ca2+ from internal stores had no effect.(ABSTRACT TRUNCATED AT 250 WORDS)

Aniline Compounds↗
PubMed1993

Volume-activated Rb+ transport in astrocytes in culture.

The involvement of K+ on the volume regulatory process in astrocytes was investigated by characterizing the hyposmolarity-induced efflux of K+ using 86Rb as a tracer. About 70 and 30% of the intracellular content of 86Rb was released after reductions in osmolarity from 320 to 160 or 220 mosM, respectively, during the time in which cells exhibit a volume regulatory response subsequent to swelling. No significant increase in 86Rb efflux was observed with lower reductions in osmolarity. The 86Rb efflux was Ca2+ independent and insensitive to temperature. It was inhibited by furosemide but not by bumetanide and was unaffected when nitrate, but not gluconate, replaced intracellular Cl-. The efflux was markedly inhibited by quinidine and by 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid. Quinidine also prevented the volume regulatory decrease of cells, and this effect was overcome when a large cation permeability was imposed by gramicidin. In isosmotic conditions 86Rb efflux was not activated by N-ethylmaleimide, but this drug strongly inhibited the hyposmolarity-activated release. These findings suggest that 86Rb efflux from astrocytes associated to cell swelling is not mediated by an electroneutral cotransporter and rather favor the implication of a conductive exit pathway that may be a Ca(2+)-independent K+ channel.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1992

Changes in taurine transport evoked by hyperosmolarity in cultured astrocytes.

Cultured astrocytes grown chronically (1-3 days) in medium made hyperosmotic (450 mOsm) with NaCl or sucrose showed an increase in taurine concentration from 294 to 501 nmol/mg protein in NaCl and to 382 nmol/mg protein in sucrose. The effect of hyperosmolarity on taurine uptake and release was examined to investigate whether or not changes in these processes may account for the increase observed in cell taurine content. Hyperosmolarity significantly affected the two components of taurine uptake (i.e., the Na(+)-dependent and the diffusional component). The Vmax of the Na(+)-dependent, active transport increased 50%, whereas no change was observed in the Km. The diffusion coefficient was markedly decreased by hyperosmolarity, being 2.2 x 10(-4) and 6.6 x 10(-6) ml/min/mg protein in isosmotic and hyperosmotic conditions, respectively, indicating a blockade of the leak pathway. These changes in the active and passive components of taurine transport were opposite to those induced by hyperosmolarity. The effect of hyperosmolarity increasing cell taurine content was insensitive to cycloheximide and colchicine. The basal efflux of taurine from astrocytes also decreased in cells exposed to hyperosmotic medium, indicating that alterations in both influx and efflux of taurine are involved in the mechanism responsible for the increase in taurine levels induced by hyperosmolarity in astrocytes.

Amino Acids↗