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Y X Li

Publications and source records attributed to Y X Li.

At least 91 records · Page 5Linked to original sources
PubMed1996

GABA stimulates chemotaxis and chemokinesis of embryonic cortical neurons via calcium-dependent mechanisms.

During rat cortical development, when neurons migrate from the ventricular zone to the cortical plate, GABA localizes within the target destinations of migratory neurons. At this time, cells in germinal zones and along migratory pathways express GABA receptor subunit transcripts, implying that in vivo, GABA may be a chemoattractant. We used an in vitro strategy to study putative chemotropic effects of GABA on embryonic rat cortical cells. GABA stimulated neuronal migration in vitro at embryonic day 15 (E15). From E16 onward, two concentration ranges (fM and microM) induced motility. Femtomolar GABA primarily stimulated chemotaxis (migration along a chemical gradient), whereas micromolar GABA predominantly initiated chemokinesis (increased random movement). These effects were mimicked by structural analogs of GABA with relative specificity at GABAA (muscimol), GABAB (R-baclofen), and GABAC (trans- or cis-4-aminocrotonic acid) receptors. Antagonists of GABAB (saclofen) and GABAC (picrotoxin) receptors partially inhibited responses to both femto- and micromolar GABA; however, only responses to femtomolar GABA were partially blocked by bicuculline, a well established antagonist of GABA at GABAA receptors. Hence, chemotactic responses to femtomolar GABA seem to involve all three classes of GABA receptor proteins, whereas chemokinetic responses to micromolar GABA involve GABAB and GABAC receptor proteins. GABA-induced motility was blocked by loading the cells with the Ca(2+)-chelating molecule bis(2-aminophenoxy)ethane-N,N,N',N'-tetra-acetic acid, suggesting that intracellular Ca2+ mediates GABA-induced cell movement. Optical recordings of cells loaded with Ca2+ indicator dye revealed that both femto- and micromolar GABA evoked increases in intracellular Ca2+. Thus, GABA-stimulated increases in intracellular Ca2+ may mediate both chemotactic and chemokinetic responses in embryonic cortical cells.

Animals↗
PubMed1996

Modeling N-methyl-D-aspartate-induced bursting in dopamine neurons.

Burst firing of dopaminergic neurons of the substantia nigra pars compacta can be induced in vitro by the glutamate agonist N-methyl-D-aspartate. It has been suggested that the interburst hyperpolarization is due to Na+ extrusion by a ouabain-sensitive pump [Johnson et al. (1992) Science 258, 665-667]. We formulate and explore a theoretical model, with a minimal number of currents, for this novel mechanism of burst generation. This minimal model is further developed into a more elaborate model based on observations of additional currents and hypotheses about their spatial distribution in dopaminergic neurons [Hounsgaard (1992) Neuroscience 50, 513-518; Llinás et al. (1984) Brain Res. 294, 127-132]. Using the minimal model, we confirm that interaction between the regenerative, inward N-methyl-D-aspartate-mediated current and the outward Na(+)-pump current is sufficient to generate the slow oscillation (approximately 0.5 Hz) underlying the burst. The negative-slope region of the N-methyl-D-aspartate channel's current-voltage relation is indispensable for this slow rhythm generation. The time-scale of Na(+)-handling determines the burst's slow frequency. Moreover, we show that, given the constraints of sodium handling, such bursting is best explained mechanistically by using at least two spatial, cable-like compartments: a soma where action potentials are produced and a dendritic compartment where the slow rhythm is generated. Our result is consistent with recent experimental evidence that burst generation originates in distal dendrites [Seutin et al. (1994) Neuroscience 58, 201-206]. Responses of the model to a number of electrophysiological and pharmacological stimuli are consistent with known responses observed under similar conditions. These include the persistence of the slow rhythm when the tetrodotoxin-sensitive Na+ channel is blocked and when the soma is voltage-clamped at -60 mV. Using our more elaborate model, we account for details of the observed frequency adaptation in N-methyl-D-aspartate-induced bursting, the origin of multiple spiking and bursting mechanisms, and the interaction between two different bursting mechanisms. Besides reproducing several well established firing patterns, this model also suggests that new firing modes, not yet recorded, might also occur in dopaminergic neurons. This model provides mechanistic insights and explanations into the origin of a variety of experimentally observed membrane potential firing patterns in dopaminergic neurons, including N-methyl-D-aspartate-induced bursting and its dendritic origin. Such a model, capable of reproducing a number of realistic behaviors of dopaminergic neurons, could be useful in further studies of the basal ganglia-thalamocortical motor circuit. It may also shed light on bursting that involves N-methyl-D-aspartate channel activity in other neuron types.

Algorithms↗
PubMed1996

[Changes in myosin isoenzymes composition and the maximum shortening velocity in hypertrophic left ventricular muscle of rats].

Changes in unloaded maximum shortening velocity (Vmax) and myosin isoenzymes (MI) composition of rat left ventricular muscle were examined in the 8-week or 16-week Goldblatt hypertensive (H8, H16) and hypertension-regressive rats (R8). The Vmax was estimated by extrapolation to zero afterload from the tension-velocity curve of left ventricular papillary muscle, while the MI composition (V1, V2 and V3) was separated by polyacrylamide gel electrophoresis and determined by densitometry. The results showed that: (1) A slow age-dependent shift to V3 and a decrease in Vmax were observed in 16- and 24-week-old rats (S8, S16), in which V1/V3 ratio was decreased respectively by 38.9% and 61.0% and Vmax was decreased respectively by 8.3% and 13.3% when compared with that of the 8-week-old rats (S0). (2) There was a significant decrease in V1/V3 ratio and Vmax in 8-week (H8) and 16-week (H16) hypertension induced hypertrophic left ventricular muscle as evidenced by the fact that the V1/V3 ratio decreased by 84.4% and 93.5% and Vmax decreased by 33.3% and 48.3% in H8 and H16 as compared with that of the control rats (S0). (3) There was a partial recovery in Vmax and V1/V3 ratio in (R8) group rats. (4) The Vmax was positively correlated with the level of V1 (r = 0.9215, P < 0.01) and negatively with the level of V3 (r = 0.9071, P < 0.01) as analyzed in all the six experimental groups of a total of 48 rats (S0, S8, S16, H8, H16, R8). In conclusion, a significant shift of the myosin isoenzymes towards low ATPase activity V3 might be the biochemical mechanism responsible, at least in part, for the decrease in maximum shortening velocity in the hypertrophic left ventricular muscle induced by pressure overload.

Animals↗
PubMed1995

Spontaneous electrical and calcium oscillations in unstimulated pituitary gonadotrophs.

Single pituitary cells often fire spontaneous action potentials (APs), which are believed to underlie spiking fluctuations in cytosolic calcium concentration ([Ca2+]i). To address how these basal [Ca2+]i fluctuations depend on changes in plasma membrane voltage (V), simultaneous measurements of V and [Ca2+]i were performed in rat pituitary gonadotrophs. The data show that each [Ca2+]i spike is produced by the Ca2+ entry during a single AP. Using these and previously obtained patch-clamp data, we develop a quantitative mathematical model of this plasma membrane oscillator and the accompanying spatiotemporal [Ca2+]i oscillations. The model demonstrates that AP-induced [Ca2+]i spiking is prominent only in a thin shell layer neighboring the cell surface. This localized [Ca2+]i spike transiently activates the Ca2(+)- dependent K+ current resulting in a sharp afterhyperpolarization following each voltage spike. In accord with experimental observations, the model shows that the frequency and amplitude of the voltage spikes are highly sensitive to current injection and to the blocking of the Ca(2+)-sensitive current. Computations also predict that leaving the membrane channels intact, the firing rate can be modified by changing the Ca2+ handling parameters: the Ca2+ diffusion rate, the Ca2+ buffering capacity, and the plasma membrane Ca2+ pump rate. Finally, the model suggests reasons that spontaneous APs were seen in some gonadotrophs but not in others. This model provides a basis for further exploring how plasma membrane electrical activity is involved in the control of cytosolic calcium level in unstimulated as well as agonist-stimulated gonadotrophs.

Action Potentials↗
PubMed1995

InsP3-induced Ca2+ excitability of the endoplasmic reticulum.

Oscillations in intracellular Ca2+ can be induced by a variety of cellular signalling processes (Woods et al., 1986; Berridge 1988; Jacob et al., 1988) and appear to play a role in secretion (Stojilković et al., 1994), fertilization (Miyazaki et al., 1993), and smooth muscle contraction (Iino and Tsukioka, 1994). Recently, great progress has been made in understanding the mechanisms involved in a particular class of Ca2+ oscillation, associated with the second messenger inositol 1,4,5-trisphosphate (InsP3) (Berridge, 1993). Working in concert with intracellular Ca2+, InsP3 controls Ca2+ release via the InsP3 receptor in the endoplasmic reticulum (ER) (Berridge and Irvine, 1989). The IP3 receptor is regulated by its coagonists InsP3 and Ca2+, which both activate and inhibit Ca2+ release (Finch et al., 1991; Bezprozvanny et al., 1991; De Young and Keizer, 1992). These processes, together with the periodic activation of Ca2+ uptake into the ER, have been identified as key features in the mechanism of InsP3-induced Ca2+ oscillations in pituitary gonadotrophs (Li et al., 1994), Xenopus laevis oocytes (Lechleiter and Clapham, 1992; Atri et al., 1993), and other cell types (Keizer and De Young, 1993). Earlier discussions and models of InsP3-induced Ca2+ oscillations focused on the nature and number of internal releasable pools of Ca2+ (Goldbeter et al., 1990; Swillens and Mercan, 1990; Somogyi and Stucki, 1991), the importance of oscillations in InsP3 (Meyer and Stryer, 1988), and other issues not based on detailed experimental findings in specific cells types.

Animals↗
PubMed1995

Ca2+ excitability of the ER membrane: an explanation for IP3-induced Ca2+ oscillations.

Recent research dealing with experiments and theoretical models of Ca2+ excitability of the endoplasmic reticulum (ER) membrane induced by inositol 1,4,5-trisphosphate (IP3) is reviewed. Ca2+ excitability refers to the ability of a small increment of cytoplasmic Ca2+ concentration ([Ca2+]i) to trigger a large [Ca2+]i pulse or oscillations. Such nonlinear regenerative behavior is conferred by the existence of IP3 channels and Ca(2+)-ATPase transporters on the ER membrane, which extends throughout the cytoplasm. Ca2+ excitability resembles the plasma membrane electrical excitability of neurons and other cells: it is driven by the ionic concentration gradient across the ER membrane (higher Ca2+ concentration inside the ER); each [Ca2+]i spike partially consumes the prestored energy that is reestablished through ATP-dependent active transport; and [Ca2+]i, the excitation variable, controls the nonlinear dynamic release rate of ER Ca2+. This review focuses on the kinetic models based on these features and on experiments dealing with the kinetic properties of [Ca2+]i-dependent gating of the IP3 receptor channel. We summarize evidence in favor of two roles for [Ca2+]i in gating the channel's opening: activation at a rapid time scale and inactivation on a slower time scale. Exploiting an analogy to the well-known Hodgkin-Huxley model for neuronal electrical excitability, we show how Ca2+ excitability of the ER membrane can be explained by these gating properties combined with the ER Ca2+ pump activity. The theory's ability to predict is illustrated by comparing calculated with experimental [Ca2+]i responses for pituitary gonadotrophs under various stimulus conditions.

Animals↗
PubMed1994

Equations for InsP3 receptor-mediated [Ca2+]i oscillations derived from a detailed kinetic model: a Hodgkin-Huxley like formalism.

The nine-variable De Young-Keizer model (1992) for [Ca2+]i oscillations mediated by InsP3 receptor channels in endoplasmic reticulum (ER) membrane is analyzed and reduced to a two-variable system. The different time scales in the three basic channel gating processes, namely InsP3 regulation, Ca2+ activation, and Ca2+ inactivation, are revealed and characterized. The method of multiple scales is used in solving the equations on a succession of faster time scales and reducing them to a 2D system. The reduced system, (Vcy/fcy) dC/dt = -P1P3Rm3 infinity h3(C-C0)-PL(C-C0)-Jpump(C); dh/dt = (h infinity-h)/tau h, is analogous in form to the Hodgkin-Huxley equations for plasma membrane electrical excitability. [Ca2+]i dynamics in this model thus involve ER membrane-associated excitability. The reduced system has a bifurcation diagram almost identical to that of the original system and retains the most important dynamic features of the latter. The analysis also shows that the reduced system becomes simpler when the different gating processes are more independent from each other, i.e. when the rates for Ca2+ binding at the site associated with one gating process are independent of occupancy at the other two binding sites. Assuming further that binding of InsP3 does not depend on Ca2+ occupancy at the inactivation site, we obtain a "minimal" form yet retain significant ability to reproduce experimental observations.

Calcium↗
PubMed1994

Calcium oscillations in pituitary gonadotrophs: comparison of experiment and theory.

We have developed a mathematical model that describes several aspects of agonist-induced Ca2+ signaling in single pituitary gonadotrophs. Our model is based on fast activation of the inositol 1,4,5-trisphosphate (InsP3) receptor Ca2+ channels at low free cytosolic Ca2+ concentration ([Ca2+]i) and slow inactivation at high [Ca2+]i. Previous work has shown that these gating properties, when combined with a Ca(2+)-ATPase, are sufficient to generate simulated Ca2+ oscillations. The Hodgkin-Huxley-like description we formulate here incorporates these different gating properties explicitly and renders their effects transparent and easy to modulate. We introduce regulatory mechanisms of channel opening which enable the model, both in the absence and in the presence of Ca2+ entry, to give responses to a wide range of agonist doses that are in good agreement with experimental findings, including subthreshold responses, superthreshold oscillations with frequency determined by [InsP3], and nonoscillatory "biphasic" responses followed occasionally by small-amplitude oscillations. A particular added feature of our model, enhanced channel opening by reduced concentration of Ca2+ in the lumen of the endoplasmic reticulum, allows oscillations to continue during pool depletion. The model predicts that ionomycin and thapsigargin can induce oscillations with basal [InsP3] and zero Ca2+ entry, while Ca2+ injection cannot. Responses to specific pairings of sub- or superthreshold stimuli of agonist, ionomycin, and thapsigargin are also correctly predicted. Since this model encompasses a wide range of observed dynamic behaviors within a single framework, based on well-established mechanisms, its relevance should not be restricted to gonadotrophs.

Calcium↗
PubMed1994

[The roles of protein kinase C and Na(+)-H+ exchange in the angiotensin II-induced hypertrophic response].

In order to study the roles of protein kinase C (PKC) and Na(+)-H+ exchange in [Sar1] ANG II-induced hypertrophic response, 3H-Leucine incorporation into serum-free cultured neonatal rat cardiomyocytes were interfered with staurosporine, phorbol 12-myristate 13-acetate (PMA) or amiloride. With [Sar1] ANG II, a dose dependent augmentation of protein synthesis was observed. Activator of PKC (PMA) could also accelerate the rate of protein synthesis to a significant extent, while pretreatment of cardiac myocytes with PKC inhibitor (staurosporine) or Na(+)-H+ exchanger inhibitor (amiloride) would block [Sar1] ANG II-induced increase 3H-Leucine incorporation. These results demonstrated that activation of PKC and Na(+)-H+ exchange might mediate the hypertrophic response initiated by [Sar1] ANG II. However, inhibition of cellular Na(+)-H+ exchange did not affect the increase in protein synthesis due to activation of PKC, a finding indicating that the two resorts to regulate cell growth are mediated by independent pathways.

Alkaloids↗
PubMed1994

The role of monosialoganglioside GM1 in LTP-induction in rat hippocampal slices.

The effect of monosialoganglioside GM1 of different doses on the long-term potentiation (LTP) of synaptic transmission has been studied in the CA1 region of rat hippocampal slices, and the possible role that calcium ion and NMDA receptor play has also been investigated. The results reveal that larger magnitude of LTP is induced in hippocampal slices pre-incubated with GM1. The dose-response curve appears in diphase, and the largest magnitude of LTP has been obtained at the GM1 concentration of 50 mg/L in incubation ACSF. Moreover, the magnitude of LTP induced from the slices pre-incubated with GM1 at lower calcium ion concentration is similar to that obtained from the control slices at normal calcium ion concentration. Under higher calcium ion concentration, the enhancing effect of GM1 on LTP seems relatively feeble. After NMDA receptors were blocked, no enhancing effect of GM1 was observed. The mechanism of GM1 action on LTP is discussed.

2-Amino-5-phosphonovalerate↗
PubMed1994

[Experimental study on huogu granule in treating aseptic necrosis of femur head].

The pharmacological activities and acute toxicological activities of Huogu Granule (HGG) on the basis of clinical treatment of aseptic necrosis of head of femur were studied. It was observed that medium and high dose of HGG could reduce the inflammatory reaction, inhibit the enhancement of the capillary permeability and decrease the frequency of body twisting obviously in mice. All of these were significantly better than that of normal saline control group (P < 0.05, P < 0.01). It showed that HGG had a noticeable anti-inflammatory and analgesic effect. Also HGG was found to improve the absorption of serum Ca, P and the development of femoral growth in chicken embryo. LD50 could not be determined in acute toxicity test, although the dose of 50g/kg.24hr was about 166.7 times of the dosage used for human adult clinically. It demonstrated that the acute toxicity of HGG is negligible.

Animals↗
PubMed1994

Gene transplantation: combined antisense inhibition and gene replacement strategies.

Optimal gene replacement protocols would include both inhibition of the endogenous gene and overexpression of the preferred (or mutant) gene. We have developed a novel gene transfer method to test whether antisense-resistant genes (designed by deletion of antisense RNA target sequences) can replace the function of endogenous genes. Immunoprecipitation studies demonstrated that inducible anti-fos RNA (antisense directed against the c-fos gene) reduces endogenous c-fos expression by 90%, but did not affect the transfected antisense-resistant mutant c-fos genes. Cell growth studies demonstrated that full-length and minimally truncated c-fos expression vectors could restore serum-induced DNA synthesis but that C-terminally truncated Fos mutants including FBR v-fos could not. Transcriptional studies demonstrate that the endogenous c-fos protein contributes to AP-1 activity and normally suppresses regulated SRE (serum response element) activity. This "gene transplant" method for inhibition of endogenous genes and replacement with preferred genes has implications for gene therapy of hereditary hematologic disorders and for the correction or "repair" of oncogenes or tumor suppressor genes in leukemias and lymphomas.

Blood Physiological Phenomena↗
PubMed1993

Subcellular distribution of Ca2+ release channels underlying Ca2+ waves and oscillations in exocrine pancreas.

Agonists trigger Ca2+ waves and oscillations in exocrine gland cells. Our confocal Ca2+ imaging revealed three distinct phases during the Ca2+ waves in the rat pancreatic acinar cell. Rises in Ca2+ concentration were initiated at a small trigger zone, or T zone, in the granular area; then, Ca2+ waves rapidly spread within the area and, at high agonist concentrations, propagated slowly toward the basal pole. Injection of inositol 1,4,5-trisphosphate (IP3) or Ca2+ from patch pipettes demonstrated the presence of high sensitivity IP3 receptors at the T zone, Ca(2+)-induced Ca2+ release channels in the granular area, and low sensitivity IP3 receptors in the basal area. The IP3 receptors at the T zone appeared to generate autonomous Ca2+ spikes and to initiate patterned Ca2+ oscillations. Thus, heterogeneous cytosolic localization of Ca2+ release channels plays a key role in Ca2+ waves and oscillations.

Acetylcholine↗
PubMed1993

Agonist-induced localized Ca2+ spikes directly triggering exocytotic secretion in exocrine pancreas.

We investigated how agonist-induced patterned rises in cytosolic Ca2+ concentration ([Ca2+]i) regulate exocytotic secretion in the rat pancreatic acinar cell. The distribution of [Ca2+]i was visualized with a confocal microscope, which revealed that a Ca2+ ionophore, A23187, induced slow and homogeneous [Ca2+]i rises, while acetylcholine (ACh) always triggered primary Ca2+ spikes at the granular area which bears secretory granules. Secretion was monitored by measuring capacitance with the patch clamp method. Errors in the estimates of membrane capacitance (C) due to changes in conductance (G) were experimentally as well as theoretically evaluated to be one-tenth of the actual signals. We found that A23187 raised G without changing C at a low concentration, while it triggered asynchronous rises in G and C with lags in C, at a high concentration. By contrast, ACh triggered simultaneous rapid rises in G and C. Our results support the hypothesis that exocytotic secretion is less sensitive to Ca2+ than to ion channels and is directly caused by agonist-induced primary Ca2+ spikes at the granular area. It is therefore suggested that spatio-temporal patterns of Ca2+ oscillations could play a key role in exocytotic secretion from the exocrine acinar cell.

Acetylcholine↗
PubMed1993

Relationship between the characteristics of immunopathological expression of hepatitis C virus antigen and hepatocytic injury.

Biopsied liver tissues from 352 cases were tested for hepatitis C virus (HCVAg) with improved PAP immunohistologic chemical method. Furthermore, corresponding seroantibody to hepatitis C virus was also tested. The total HCVAg positive rate was 9.1%. The HCVAg positive rate in chronic persistent hepatitis (CPH) was 5%. The HCVAg positive rate in chronic active hepatitis (CAH) was 11.2%. The HCVAg positive rate raised gradually along with the severity of hepatocytic injury. HCVAg may be seen in necrotic liver cells exfoliating into the liver sinus, indicating a close relationship between HCVAg and hepatocytic injury. Expression of HCVAg was mostly of the nucleus type in CPH cases and was mostly of the plasma type in CAH cases. The periphery of nucleus type-expressed positive cells generally had no marked inflammatory cell infiltration. The periphery of plasma type-expressed positive cells had a certain amount of inflammatory cell infiltration. Along with the severity of hepatocytic injury, HCVAg expressed itself in a positive correlation according to the nucleus and plasma types. The HCVAg positive cells were located mostly in the lobular peripheral band and rarely located in the venoperipheral band. It was possible that this had some relation with the lobular microcirculation of blood and blood supply. In this study, there was no obvious correlation between the HCVAg positive rate in hepatic tissues and the anti-HCV positive rate in sera. Neither the patients with HCVAg positive liver tissues nor the patients with seropositive anti-HCV had any history of blood transfusion and the use of blood products.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1993

[Defects in insulin binding, internalization and degradation of monocytes from patients with non-insulin-dependent diabetes mellitus].

A14-125I-monoiodoinsulin binding to, internalization into, and degradation inside human circulating monocytes, a cell type widely used for insulin receptor studied, from normal(32) and weight, age-matched NIDDM(31) subject were studied. The specific insulin binding, internalization and degradation were decreased in the cells from NIDDM compared to that in the cells from normal subjects. Scatchard plot analysis of these group data reveal that cells from the normal control subject contain 28,000 receptor sites per cell, while monocytes from the NIDDM patients contain 15,000 sites per cell. These results indicate that the decreased abilities of insulin binding, internalization and degradation of target cells may play a role in the cellular resistance to insulin that occurs in the NIDDM.

Adult↗