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S L Xia

Publications and source records attributed to S L Xia.

17 recordsLinked to original sources
PubMed2001

Apical membrane of native OMCD(i) cells has nonselective cation channels.

The purpose of this study was to examine cation channel activity in the apical membrane of the outer medullary collecting duct of the inner stripe (OMCD(i)) using the patch-clamp technique. In freshly isolated and lumen-opened rabbit OMCD(i), we have observed a single channel conductance of 23.3 +/- 0.6 pS (n = 17) in cell-attached (c/a) patches with high KCl in the bath and in the pipette at room temperature. Channel open probability varied among patches from 0.06 +/- 0.01 at -60 mV (n = 5) to 0.31 +/- 0.04 at 60 mV (n = 6) and consistently increased upon membrane depolarization. In inside-out (i/o) patches with symmetrical KCl solutions, the channel conductance (22.8 +/- 0.8 pS; n = 10) was similar as in the c/a configuration. Substitution of the majority of Cl- with gluconate from KCl solution in the pipette and bath did not significantly alter reversal potential (E(rev)) or the channel conductance (19.7 +/- 1.1 pS in asymmetrical potassium gluconate, n = 4; 21.4 +/- 0.5 pS in symmetrical potassium gluconate, n = 3). Experiments with 10-fold lower KCl concentration in bath solution in i/o patches shifted E(rev) to near the E(rev) of K+. The estimated permeability of K+ vs. Cl- was over 10, and the conductance was 13.4 +/- 0.1 pS (n = 3). The channel did not discriminate between K+ and Na+, as evidenced by a lack of a shift in the E(rev) with different K+ and Na+ concentration solutions in i/o patches (n = 3). The current studies demonstrate the presence of cation channels in the apical membrane of native OMCD(i) cells that could participate in K+ secretion or Na+ absorption.

Animals↗
PubMed2001

Increased CO(2) stimulates K/Rb reabsorption mediated by H-K-ATPase in CCD of potassium-restricted rabbit.

Apical H-K-ATPase in the cortical collecting duct (CCD) plays an important role in urinary acidification and K reabsorption. Our previous studies demonstrated that an H-K-ATPase mediates, in part, Rb reabsorption in rabbit CCD (Zhou X and Wingo CS. Am J Physiol Renal Fluid Electrolyte Physiol 263: F1134-F1141, 1992). The purpose of these experiments was to examine using in vitro microperfused CCD from K-restricted rabbits 1) whether an acute increase in PCO(2) and, presumably, intracellular acidosis stimulate K absorptive flux; and 2) whether this stimulation was dependent on the presence of a functional H-K-ATPase. Rb reabsorption was significantly increased after exposure to 10% CO(2) in CCD, and this effect was persistent for the entire 10% CO(2) period, whereas 10 microM SCH-28080 in the perfusate totally abolished the stimulation of Rb reabsorption by 10% CO(2). After stimulation of Rb reabsorption by 10% CO(2), subsequent addition of 0.1 mM methazolamide, an inhibitor of carbonic anhydrase, failed to affect Rb reabsorption. However, simultaneous exposure to 10% CO(2) and methazolamide prevented the stimulation of Rb reabsorption. Treatment with the intracellular calcium chelator MAPTAM (0.5 microM) inhibited the stimulation of Rb reabsorption by 10% CO(2). Similar inhibition was also observed in the presence of either a calmodulin inhibitor, W-7 (0.5 microM), or colchicine (0.5 mM), an inhibitor of tubulin polymerization. In time control studies, the perfusion time did not significantly affect Rb reabsorption. We conclude the following: 1) stimulation of Rb reabsorption on exposure to 10% CO(2) is dependent on the presence of a functional H-K-ATPase and appears to be regulated in part by the insertion of this enzyme into the apical plasma membrane by exocytosis; 2) insertion of H-K-ATPase requires changes in intracellular pH and needs a basal level of intracellular calcium concentration; and 3) H-K-ATPase insertion occurs by a microtubule-dependent process.

Absorption↗
PubMed2000

Activation of H(+)-K(+)-ATPase by CO(2) requires a basolateral Ba(2+)-sensitive pathway during K restriction.

We studied the activation of H(+)-K(+)-ATPase by CO(2) in the renal cortical collecting duct (CCD) of K-restricted animals. Exposure of microperfused CCD to 10% CO(2) increased net total CO(2) flux (J(t CO(2))) from 4.9 +/- 2.1 to 14.7 +/- 4 pmol. mm(-1). min(-1) (P < 0. 05), and this effect was blocked by luminal application of the H(+)-K(+)-ATPase inhibitor Sch-28080. In the presence of luminal Ba, a K channel blocker, exposure to CO(2) still stimulated J(t CO(2)) from 6.0 +/- 1.0 to 16.8 +/- 2.8 pmol. mm(-1). min(-1) (P < 0.01), but peritubular application of Ba inhibited the stimulation. CO(2) substantially increased (86)Rb efflux (a K tracer marker) from 93.1 +/- 23.8 to 249 +/- 60.2 nm/s (P < 0.05). These observations suggest that during K restriction 1) the enhanced H(+)-K(+)-ATPase-mediated acidification after exposure to CO(2) is dependent on a basolateral Ba-sensitive mechanism, which is different from the response of rabbits fed a normal-K diet, where activation of the H(+)-K(+)-ATPase by exposure to CO(2) is dependent on an apical Ba-sensitive pathway; and 2) K/Rb absorption via the apical H(+)-K(+)-ATPase exits through a basolateral Ba-sensitive pathway. Together, these data are consistent with the hypothesis of cooperation between H(+)-K(+)-ATPase-mediated acidification and K exit pathways in the CCD that regulate K homeostasis.

Animals↗
PubMed1999

Cation channels of the rabbit outer medullary collecting duct.

The outer medullary collecting duct (OMCD) produces high rates of proton secretion, and previous data show a significant role for an H+,K+-ATPase in luminal acidification. This mechanism of acidification requires proton secretion to occur in exchange for potassium absorption. Because net transport of potassium is small in the OMCD, pathways for potassium secretion would appear essential to explain the contribution of an H+,K+-ATPase under potassium-replete conditions. A significant issue is whether a potassium exit pathway (i.e., cation channel) is present at the apical and/or the basolateral membrane. In this report, using patch-clamp techniques to examine single channel conductances from the native rabbit OMCD, we show that the apical membrane has potassium permeable cation channels.

Animals↗
PubMed1998

Chloride transport by the rabbit cortical collecting duct: dependence on H,K-ATPase.

The rabbit cortical collecting duct (CCD) exhibits the capacity for active chloride absorption when basolateral Na-K-ATPase is inhibited by ouabain. The present studies examine the contribution of H,K-ATPase to this ouabain-insensitive Cl absorption and to related ion fluxes. Rabbits were fed a KCl-rich diet with no measurable Na for 4 to 13 d before isolation of the CCD for microperfusion. Application of peritubular ouabain (0.1 mM) significantly increased (P < 0.001) net luminal absorptive chloride flux (J(N)Cl) without an effect on lumen-to-bath isotopic 36Cl flux (J(lb)Cl). The H,K-ATPase inhibitor Sch 28080 (1 to 10 microM) abolished ouabain-insensitive J(N)Cl, but transepithelial voltage (V(T)) was not significantly affected. The contribution of H,K-ATPase activity on active Cl flux (J(A)Cl) and passive Cl flux (J(P)Cl) was also assessed. Ouabain significantly increased J(A)Cl and Sch 28080 inhibited J(A)Cl, but J(P)Cl was not affected by Sch 28080. To assess the contribution of changes in net bicarbonate flux (JtCO2) to changes in J(N)Cl, JtCO2 was measured under identical conditions as for J(N)Cl. Ouabain significantly increased JtCO2, and this ouabain-insensitive bicarbonate flux was inhibited by Sch 28080 without significantly affecting V(T). To assess the possibility that the CCD may possess mechanisms for neutral salt absorption, lumen-to-bath 86Rb efflux (K(Rb)), and 22Na efflux (K(Na)) were also measured. Ouabain significantly increased K(Rb), and Sch 28080 inhibited this ouabain-insensitive K(Rb). Furthermore, Sch 28080 and A80915a (a structurally distinct H,K-ATPase inhibitor) significantly inhibited K(Na) in the presence of 1 mM luminal amiloride. These observations suggest that, in addition to potassium, sodium can be transported via the H,K-ATPase. Although the CCD contains more than one cell population, the data could be fitted very well to the function of the B-type intercalated cell. A cell model is proposed for the hypothesis that ouabain-insensitive chloride absorption is mediated by the parallel operation of an apical H,K-ATPase with an apical Cl-HCO3 exchanger and that the H,K-ATPase can function, under certain conditions, as a mechanism of Na absorption.

Animals↗
PubMed1998

Synergistic receptor-activated calcium increases in single nonpigmented epithelial cells.

PURPOSE: To determine whether single nonpigmented ciliary body cells contain the signaling mechanism to produce synergistic drug-activated increases in Ca2+, or whether these responses are produced cooperatively by interaction among groups of cells. METHODS: Suspensions of single nonpigmented cells were plated onto soft collagen gels. Fura-2 fluorescence ratio imaging was used to examine receptor-evoked changes in intracellular Ca2+ concentration. RESULTS: Nonpigmented cells plated on soft collagen gels retained a rounded shape with membrane evaginations visible on their surface. Application of acetylcholine (10 microM) or epinephrine (1 microM) each produced small increases in intracellular Ca2+, but in combination they produced a Ca2+ increase of more than 10-fold. This synergistic Ca2+increase was a result of activation of muscarinic and alpha2-adrenergic receptors because a specific alpha2-adrenergic agonist could substitute for epinephrine in producing the response. The response could be blocked by a specific alpha2-antagonist and a muscarinic antagonist. An alpha1-agonist could not substitute for epinephrine in producing a synergistic increase nor could the synergism be blocked by alpha1- or beta-antagonists. The Ca2+ increase was largely produced by release from internal stores, because the peak amplitude of the response was nearly the same in the external solution containing a low Ca2+ concentration; however, the influx of Ca2+ into the cell was responsible for maintenance of a steady component of the Ca2+ increase during maintained drug stimulation and for refilling the internal stores. CONCLUSIONS: Single nonpigmented cells can produce synergistic increases in Ca2+ on multiple receptor activation, indicating that the mechanism of synergism does not require the interaction of multiple cells. The Ca2+ increase is a result of release from internal stores and Ca2+ entry through an as yet undefined conductance or transport system in the plasma membrane.

Acetylcholine↗
PubMed1997

Synergistic rise in Ca2+ produced by somatostatin and acetylcholine in ciliary body epithelial cells.

The purpose of these experiments was to demonstrate the presence of somatostatin receptors on the nonpigmented epithelial cells of the rabbit ciliary body and their link with intracellular Ca2+ homeostasis. Freshly excised rabbit ciliary processes and nonpigmented cell layer, explants were loaded with the fluorescent dye fura-2, and free-Ca2+ concentration ([Ca2+]i) in the nonpigmented cells was measured with fluorescence ratio imaging. The cells were continuously perfused, and drugs were added to the perfusate. Somatostatin-14 (SS14, 0.1-1.0 microM) or acetylcholine (ACh, 10 microM) applied alone produced small increases in [Ca2+]i. However, SS14 (0.1 microM) in combination with ACh (10 microM) induced a massive increase in [Ca2+]i (25.7 +/- 3.3 times the baseline level, n = 28). The dose-response curve for SS14 (in the presence of 10 microM ACh) was sigmoidal with an EC50 of 3.9 nM and Hill coefficient of 2.5, indicating the requirement for multiple SS receptor activation. Somatostatin-28 could mimic the effect of SS14, although a much higher concentration was required. Shifting the SS14 dose-response curve to the right by about two-orders of magnitude resulted in a fit to the SS28 data. The response to ACh + SS14 could not be blocked by the alpha 2-adrenergic blocker yohimbine (Yoh, 10 microM) or the A1-specific adenosinergic antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX, 1 microM). Incubation of the tissue with pertussis toxin (PTx, 1 microgram ml-1) did not alter the response to ACh alone but eliminated the synergistic effect of somatostatin. We conclude that nonpigmented epithelial cells of the rabbit ciliary body possess a novel somatostatin receptor whose activation can synergistically potentiate the rise in [Ca2+]i produced by ACh. This potentiation appears to occur via a pertussis-toxin-sensitive pathway, perhaps through Gi.

Acetylcholine↗
PubMed1996

Localized calcium signaling in multinucleated osteoclasts.

Localized intracellular Ca2+ ([Ca2+]i) pulses, fluctuations, and repetitive spikes were detected in multinucleated rabbit osteoclasts in the presence of serum and in response to calcitonin using the fluorescent calcium indicator fluo-3 and a laser scanning microscope. We observed that these [Ca2+], changes were often restricted within a region of the cell body or propagated from the initial region of occurrence to other parts of the cell body but not to all parts. These observations suggest the existence of significant barriers to Ca2+ transport between different cytoplasmic regions of the osteoclast. To further investigate this phenomenon, we mechanically perturbed different cellular regions by touching locally with a micropipette. This usually induced a local increase in cytosolic and nuclear free [Ca2+]i. In some cases there was propagation of the [Ca2+]i increase to other regions but with part of the cell body not affected. Those regions of the cell body to which the [Ca2+]i increase did not propagate had a [Ca2+]i response to a direct mechanical perturbation. Our data show that osteoclasts can have different [Ca2+]i activities in apparently equivalent cellular regions, no matter how generated. This suggests that there can be a number of spatially separate Ca2+ regulatory systems within an osteoclast cell body.

Animals↗
PubMed1995

Calcium signal induced by mechanical perturbation of osteoclasts.

Multinucleated osteoclasts from rabbit long bone, 1-6 days in culture, respond to mechanical perturbation with a transient increase of intracellular calcium concentration ([Ca2+]i), as measured with the fluorescent indicator fluo-3 on a confocal laser scanning microscope. In experiments with different extracellular calcium concentrations (from 11.8 mM to calcium-free), the incidence, the magnitude, and the duration of [Ca2+]i responses decreases with decreasing bathing [Ca2+]. Following mechanical perturbation, a thapsigargin-induced [Ca2+]i response has a lower magnitude than the thapsigargin-induced response without mechanical perturbation. In thapsigargin-pretreated osteoclasts the mechanical perturbation-induced rise in [Ca2+]i is larger and longer than in control cells. Ni2+ inhibits the incidence and decreases both the magnitude and the duration of the responses, while nifedipine, verapamil, and Gd3+ have no effect. These measurements show that rabbit osteoclasts transduce a mechanical perturbation of the cell membrane into a [Ca2+]i signal via both a calcium influx and an internal calcium release.

Animals↗
PubMed1995

[The induction and differentiation of a human teratocarcinoma cell line (PA-1) in vitro].

PA-1 cell line is derived from human ovary teratocarcinoma. When it grows in 10% fetal calf serum, it can be induced to differentiate by 10(-5) mol/L retinoic acid (RA). Some morphological changes can be observed after RA induction. By immunostaining of cultured cells, we found that the expression and distribution pattern of desmin and some extracellular matrix molecules, such as fibronectin, laminin and tenascin, had been changed after induction, and these changes were associated with the morphological changes. Cell growth study showed that RA treatment had no effects on growth and autocrine activities of PA-1 cells. These results suggest that some PA-1 cells were induced to differentiate along muscle cells pathway by RA.

Cell Transformation, Neoplastic↗
PubMed1994

Displacement and translocation of osteoblast-like cells by osteoclasts.

Rabbit osteoclasts and rabbit osteoblast-like stroma cells (OB cells) were placed onto plastic surfaces and the migration patterns of individual osteoclasts and osteoclast-OB interactions were analyzed with time-lapse recording. To induce directed migration, the cultures were exposed to an electrical field of 0.01 or 0.1 V/mm. At 0.1 V/mm, osteoclasts moved directly toward the anode in some cases, clearing OB cells from their path of migration. In other cases, osteoclasts migrated toward the anode for part of the time but then changed direction and moved toward groups of OB cells. Observations were made on osteoclasts interacting with single OB cells or small colonies and on osteoclasts interacting with OB monolayers, at both field strengths; the results were independent of field strength. There were several characteristic behaviors. With single OB cells and small OB colonies, retraction of OB cells upon contact with the osteoclast was the predominant mechanism whereby these cells begin to move out of the path of the osteoclast. A pronounced ruffling or blebbing of the OB cell membrane often followed retraction. When osteoclasts displaced OB cells that were part of a monolayer, extension of an osteoclast lamellipodium underneath the edge of the OB cell layer generally preceded partial retraction of the OB cells involved. It sometimes appeared as if the detached or partially detached OB cells were "pushed" by the osteoclast, which in some cases resulted in OB cells being moved hundreds of microns in a period of a few hours, at rates comparable to the normal speed for osteoclast migration (congruent to 100 microns/h), much faster than the normal speed for OB cells (congruent to 10 microns/h).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Propagation of a calcium pulse between osteoblastic cells.

Using rat calvaria cells in primary culture monolayers and bone-like nodules, and isolated rat osteosarcoma cells, we show via laser scanning confocal microscopy and fluorescent indicator fluo-3/AM, that mechanical perturbation of a cell results in a transient increase (pulse) of measured intracellular calcium concentration that propagates from cell to cell, even between cells connected only by a thin process. The calcium pulse does not occur in the mechanically perturbed cell in calcium-free bathing medium, nor is there pulse propagation under this condition. Halothane, which blocks gap junctions, inhibits propagation. Propagation velocity does not decrease with successive cell to cell steps. These observations suggest the existence of a self-regenerating calcium signaling mechanism that may be based on a form of calcium-induced calcium release.

Aniline Compounds↗
PubMed1992

Hormone responses of in vitro bone nodule cells: studies on changes of intracellular calcium and membrane potential in response to parathyroid hormone and calcitonin.

We used two techniques to study the responses of individual in vitro bone nodule cells to parathyroid hormone (PTH) and calcitonin (CT). These techniques are laser scanning confocal imaging with a fluorescent indicator to measure intracellular free [Ca2+], and microelectrode impalement to measure the electrical potential difference across the cell membrane. We applied these measurement techniques to cells in the top cellular layer of nodules that form in vitro in cultures of cells obtained from fetal rat calvaria. Our measurements showed a transient increase in intracellular free [Ca2+] following application of PTH or CT. The duration of the increase in fluorescent intensity following PTH application varied from about 100 to more than 300 s, and the duration following CT application was from 30 to 80 s. In some measurements we applied both hormones in sequence, and observed that some cells showed an intracellular [Ca2+] response to both hormones, while other cells apparently responded to only one or the other of the hormones, or to neither. We also observed membrane potential changes in response to PTH and to CT. The membrane potential response to CT was quite small. The time courses of these membrane potential changes consisted of a depolarizing phase lasting about 100 s (with both hormones) followed by a hyperpolarizing phase (with PTH). Control measurements using only the vehicle solutions were carried out with both techniques, producing negligible responses.

Animals↗
PubMed1991

Ba(2+)-induced action potentials in osteoblastic cells.

Trains of long-duration "action potentials" were induced by Ba2+ in osteoblast-like rat osteosarcoma cells (ROS 17/2.8), under current clamp and voltage clamp. Large depolarizing pulses were seen in microelectrode measurements at 37 degrees C following the addition of 10 or 20 mM Ba2+ to physiological bathing medium. Application of BAY K 8644 resulted in the onset of the pulses at earlier times and at more negative potentials. The pulses were blocked by nifedipine and Cd2+, but not by Ni2+. Large inward current pulses were seen in whole-cell patch technique voltage-clamp measurements at 37 degrees C in the presence of from 10 to 110 mM Ba2+ in the bathing medium. The current pulses were not seen at 22 degrees C in the presence of 110 mM Ba2+, but could be induced by BAY K 8644. These pulses were not blocked by TTX, but were blocked by nifedipine, Cd2+, Zn2+, Co2+, and by an increase in bathing [Ca2+]. The shape and frequency of the current pulses were the same as for voltage pulses under current clamp. A model that can explain these observations involves opening of L-type Ca2+ channels in a voltage-independent manner by cytosolic Ba2+ via a screening of Ca2+ from sites that produce either inactivation or a lower probability of opening in the activated state. There would be a closing of these channels at higher [Ba2+] as Ba2+ is forced onto these sites. A refractory period is also required to give repeated pulses of openings.

Action Potentials↗