Biomedical subjects

L C Wang

Publications and source records attributed to L C Wang.

At least 19 recordsLinked to original sources

PubMed1992

Central role for differential gene expression in mammalian hibernation.

Mammalian hibernators experience dramatic reductions in body temperature, metabolic rate, respiratory rate, and heart rate during hibernation. These changes are precisely controlled and reversible with only internally driven mechanisms, suggesting specific biochemical regulation. We present a model that integrates our observations of differential liver gene expression during preparation for, and maintenance of, the hibernating state, with the known phylogenetic interspersion of hibernating species in several major mammalian lineages. This model predicts a major role for the differential expression of existing mammalian genes in the biochemical regulation of hibernation.

Animals↗

PubMed1992

Specificity of a target cell-derived stop signal for afferent axonal growth.

With a novel model culture system in which afferents are co-cultured with purified populations of target neurons, we have demonstrated that a target cell within the central nervous system (CNS), the cerebellar granule neuron, poses a "stop-growing signal" for its appropriate afferents, the mossy fibers. To ask whether this stop signal is afferent specific, we co-cultured granule neurons with another cerebellar afferent system, the climbing fibers from the inferior olivary nuclei, which normally contact Purkinje neurons, and with retinal ganglion cell afferents, which never enter the cerebellum. Granule neurons do not pose a stop signal to either of these afferents. In contrast to pontine mossy afferents that grow well on laminin and showed reduced outgrowth on granule neurons, both olivary and retinal fibers displayed similar growth on laminin alone or on granule neurons. In addition, each afferent showed different degrees of fasciculation and growth cone morphology on laminin. Thus, the growth arrest signal sent by granule neurons is specifically recognized by their appropriate afferents. Moreover, these three types of afferents exhibit varying growth patterns on the same noncellular and cellular substrates, implicating distinct molecular characteristics of growth regulation for different classes of neurons that would contribute to specificity of synapse formation.

Animals↗

PubMed1992

Decrease in cold tolerance of aged rats caused by the enhanced endogenous adenosine activity.

During severe cold exposure, old rats (24-28 months) were less capable of maintaining their body temperature compared to young rats (3-6 months) due to lower rate of heat production. Single injection of adenosine deaminase (AD) (converts adenosine to inosine) significantly increased thermogenesis in both young and old rats. However, doubling the dose of AD was required for optimal thermogenic response in old rats. In contrast, the similar enhancements in both thermogenesis and cold tolerance were observed in both young and old rats receiving the same optimal doses of specific adenosine receptor antagonists. These results lead to the suggestion that the lower capability of aged rats to withstand cold exposure could be due to an increase in adenosine stimulation because of the decreased endogenous AD activity rather than an increase in adenosine receptor sensitivity. This notion is further supported by the finding that the AD activity in the neck muscle, a key site for shivering thermogenesis, was significantly lower in old rats as compared to their younger counterparts before and after cold exposure.

Adenosine↗

PubMed1992

Prolonged stable hypothermia: effect on blood gases and pH in rats and ground squirrels.

Richardson's ground squirrels [body temperature (Tb) 7 degrees C] survive prolonged stable hypothermia for three times as long as do rats (Tb 19 degrees C) (72 vs. 24 h). We have examined the changes in blood gases and acid-base state to assess whether these contribute to this difference in survival time. None of the variables (measured at ambient temperature of 25 degrees C) differed significantly between rats and ground squirrels before hypothermic induction. During cooling, neither hematocrit nor plasma lactate changed significantly, but arterial and venous PO2 and PCO2 increased and arterial and venous pH decreased in both groups. During prolonged hypothermia, hematocrit increased significantly in rats (58.8 +/- 1.7% at 24 h) but not in ground squirrels (39.1 +/- 1.0% at 72 h). Both species maintained stable arterial blood gases but showed decreased venous PO2; arterial and venous pH decreased significantly with time in both species in conjunction with increased plasma lactate. These patterns of decreased venous PO2 and increased plasma lactate suggest that reduced tissue oxygenation occurs during hypothermia. This happens earlier in rats at a Tb of 19 degrees C than in ground squirrels at a Tb of 7 degrees C, possibly as a result of increased hematocrit in hypothermic rats. Remedial measures directed at improving tissue O2 delivery may therefore prolong the hypothermic survival of rats.

Animals↗

PubMed1992

Temperature dependence of electrophysiological properties of guinea pig and ground squirrel myocytes.

The effects of changing temperature on the electrophysiology of isolated cardiac myocytes of the guinea pig and Richardson's ground squirrel were studied by patch-clamp techniques. In cells from both species, the resting membrane potential declined on cooling from 36 to 12 degrees C by approximately 6 mV. The duration of the plateau of the action potential in guinea pig cells increased monotonically on cooling. In contrast, the action potential of ground squirrel cells showed a biphasic response, increasing in duration from 36 to 24 degrees C and then decreasing on cooling from 24 to 12 degrees C. From voltage-clamp studies, the properties of L-type calcium currents (ICa) on cooling were compared in the two species and were found to be similar: In both cases, ICa decreased in amplitude from approximately 2 nA peak current at 36 degrees C to less than 400 pA at 12 degrees C. The Q10 of both the maximum amplitude and time to peak for ICa in both species was approximately 1.8. The time for half inactivation had a greater Q10 of 2.5-3. It is concluded that, surprisingly, factors affecting the resting membrane potential and properties of L-type calcium channels are not major contributors to cardiac dysfunction on cooling. Rather, it is sarcoplasmic reticulum calcium release and reuptake that are likely to be the most important cold-sensitive processes.

Animals↗

PubMed1992

[The synaptic architecture of propriospinal neurons in cultured mouse spinal cord].

An electron microscopic analysis of the synaptic architecture in propriospinal neurons of cultured fetal mouse spinal cord has been undertaken. The size of the perikarya in the cultured spinal cord represents a range from small- to medium sized neurons, which form many synapses each other. There are many axo-dendritic and axo-somatic synapses in the culture but direct dendro-dendritic apposition is rarely seen. Four morphological types of synaptic boutons, S, F, M and G are classified according to criteria used by previous investigators. The ultrastructural details available suggest that the propriospinal neurons receive synaptic input from propriospinal fibers through simple synapses. It may indicate that their impulses can be controlled only postsynaptically.

Animals↗

PubMed1991

Seasonal changes in methionine-enkephalin immunoreactivity in the brain of a hibernator, Spermophilus columbianus.

To identify the actual location of central endogenous opioid systems which may be involved in regulating the hibernation cycle, differences in the pattern of central methionine-enkephalin (Met-EK) immunoreactivity were compared between hibernating (body temperature, Tb = 7 degrees C) and non-hibernating (Tb = 37 degrees C) Columbian ground squirrels using the peroxidase-antiperoxidase technique. In non-hibernating animals, Met-EK-immunoreactive perikarya were observed in telencephalic (putamen, caudate nucleus, medial septum-diagonal band complex, amygdala) and diencephalic (periventricular hypothalamic nucleus, lateral hypothalamic area) regions, whereas immunoreactive fibers were found in the lateral septum, stria terminalis nucleus, various hypothalamic areas, arcuate nucleus, median eminence, thalamic intralaminar, periventricular nucleus and lateral habenular nucleus. Compared to the non-hibernating animal, a marked increase in the number of Met-EK-immunoreactive fibers was found in the lateral septal nucleus, the periventricular nucleus, the intralaminar thalamus and the paraventricular hypothalamus of hibernating ground squirrels. Since these changes in immunoreactivity were not observed in the artificially induced hypothermic ground squirrels (Tb = 7 degrees C), it is unlikely that the dissimilarity in immunoreactivity between animals from different hibernating phases is due to differences in their Tb. In combination with our previous studies, these results tend to suggest that hibernation may be brought about by an increase in endogenous opioid activity, especially in the lateral septal region.

Animals↗

PubMed1991

Altered properties of calsequestrin and the ryanodine receptor in the cardiac sarcoplasmic reticulum of hibernating mammals.

A novel isoform of calsequestrin was identified in sarcoplasmic reticulum vesicles from myocardial tissue of two species of hibernating ground squirrel. The protein was identified as calsequestrin by its cross-reactivity with antibodies raised against bovine cardiac calsequestrin, its pH-sensitive mobility in sodium dodecylsulphate-polyacrylamide gels, staining blue with the cationic carbocyanine dye 'Stains-All', binding peroxidase-conjugated concanavalin A, its endoglycosidase F sensitivity. Its NH2-terminal amino acid sequence is similar, but not identical, to that already determined for cardiac calsequestrin. Some of the biochemical properties of this protein distinguish it from the other mammalian isoforms. It has a unique electrophoretic mobility in both alkaline and neutral sodium dodecylsulphate-polyacrylamide gel electrophoresis, it appears to have a molecular weight approximately 7% greater than that of cardiac calsequestrin from other mammalian species, and its glycosylation pattern differs. This novel form of calsequestrin is expressed in cardiac SR vesicles which possess an abnormally high number of Ca2(+)-release channel/ryanodine receptor molecules. This ryanodine receptor also shows an altered Ca2(+)-sensitivity of ryanodine binding. The divergent biophysical properties of this novel form of cardiac calsequestrin, together with the apparently atypical ryanodine receptors in the cardiac sarcoplasmic reticulum membranes may have some functional significance in the adaptive mechanisms which allow the heart to function despite the severely reduced body temperatures (to approx. 0 degree C) encountered during hibernation.

Amino Acid Sequence↗

PubMed1991

Seasonal variations in the rate and capacity of cardiac SR calcium accumulation in a hibernating species.

The rate of calcium uptake and the level of calcium accumulation was measured in cardiac muscle SR from hibernating and nonhibernating Richardson's ground squirrels. In whole heart homogenates, the rate of calcium uptake was higher (P less than 0.05) in hibernating animals than it was in active animals. Further purification of homogenates into sacroplasmic reticulum (SR) preparations showed that the hibernating animals had the highest rate of calcium uptake and the greatest level of calcium accumulation. These results could not be explained by variations in non-SR membrane contaminants nor by changes in the maximal activity or total amount of a SR marker enzyme, the Ca(2+)-ATPase. The addition of ryanodine to the calcium uptake medium increased the level of calcium accumulation in all groups by a similar amount. It is concluded that the high rate of calcium uptake by isolated cardiac SR vesicles from hibernating ground squirrels reflects the activity of the organelle in vivo, and that the ability of the ryanodine-insensitive population of SR vesicles to accumulate calcium is affected by hibernation.

Animals↗

PubMed1991

State-dependent variation in the inhibitory effect of [D-Ala2, D-Leu5]-enkephalin on hippocampal serotonin release in ground squirrels.

Accumulated evidence has suggested that increased endogenous opioid activities may facilitate the onset of hibernation either directly or possibly through modulation of other neurotransmitter systems. The seasonal change of [D-Ala2, D-Leu5]-enkephalin (DADLE), a delta receptor agonist, in modulating K+ (35 mM)-induced [3H]-5-hydroxytryptamine (5-HT) release from the hippocampal and hypothalamic slices of euthermic and hibernating Richardsons' ground squirrels was therefore investigated. DADLE (0.1-10 microM) had no effect on 5-HT release in the hypothalamic slices but elicited a dose-related inhibition on [3H]-5-HT release from the hippocampal slices of the euthermic ground squirrel. The inhibitory effect of DADLE was completely reversed by naloxone (10 microM), but not by tetrodotoxin (1 microM). In contrast, DADLE failed to alter the K(+)-induced 5-HT release from the hippocampal slices of the hibernating ground squirrel. This state-dependent reduction in responsiveness to an opioid is consistent with the hypothesis that enhanced endogenous opioid activity in the hibernating phase could lead to down regulation of the opioid receptors and minimize its inhibition on hippocampal serotonergic activity. A high 5-HT activity would inhibit midbrain reticular activating system indirectly through non-serotonergic fibers, which in turn facilitate the onset or maintenance of hibernation.

Animals↗

PubMed1991

Effect of low temperature on the cytosolic free Ca2+ in rat ventricular myocytes.

The effect of low temperature on the cytosolic free Ca2+ [( Ca2+]i) has been investigated in isolated ventricular myocytes from adult rats using the fluorescent probe Indo-1. The distribution of Indo-1 between the mitochondrial and cytoplasmic compartments was first determined in the isolated myocytes using the digitonin and Triton X-100 treatments. By subtracting the mitochondrial [Ca2+]i from the total [Ca2+]i measured with Indo-1, the average cytosolic [Ca2+]i was found to increase significantly (P less than 0.05) from 139 nM to 255 and 297 nM when the temperature was decreased from 37 degrees C to 15 degrees and 5 degrees C, respectively. A marked increase in cytosolic [Ca2+]i to a new steady state level was observed when the membrane of myocytes was depolarized by 60 mM KCI; the average magnitude of increase being 110, 243 and 186 nM, at 37 degrees, 15 degrees and 5 degrees C respectively. Our results support the hypothesis that the cardiac arrhythmia typically observed in the hypothermic rat is due to an increased cytosolic [Ca2+]i with decreasing body temperature.

Animals↗

PubMed1991

Experimental infection routes of Angiostrongylus cantonensis in mice.

Stomach intubation is the most common method used in the experimental infection of animals with Angiostrongylus cantonensis. In order to compare the effectiveness of other possible transmission methods, groups of BALB/c mice were given infective third-stage larvae of A. cantonensis by different routes including intraperitoneal or subcutaneous injections, and penetration of anal mucosa, vaginal mucosa, conjunctival mucosa, lacerated skin, unabraded skin, foot pad and tail skin, while stomach intubation was used as control. Recovery of fifth-stage larvae was higher in mice inoculated with third-stage larvae subcutaneously. Successful infections were established through all experimental transmission routes except tail skin penetration. This study suggests that oral infection may not be the only route for the transmission of human angiostrongyliasis, and subcutaneous infection may be a better method for experimental infection.

Anal Canal↗

PubMed1991

Optimal temperature in short-term hypothermic preservation of rat pancreas.

Rat pancreases were hypothermically preserved for 24 hr in University of Wisconsin preservation solution at specific temperatures (4, 7, 10, and 15 degrees C) to evaluate the effect of preservation temperature on islet yield and islet function. Following hypothermic storage, the loss of recoverable islets was highest in the 15 degrees C-preservation group, followed by that of the 4 degrees C group and significantly less in the 7 and 10 degrees C groups (p less than 0.05). Islet function, as measured by insulin output following glucose challenge in perifusion, was reduced (p less than 0.0001) in all preservation groups. The mean insulin secretion rate was lowest in the 4 degrees C group, highest in the 7 degrees C group, and intermediate in the 10 degrees C group. The islet preservation index, a parameter taking into consideration both islet yield and islet function after preservation, was more than double in the 7 and 10 degrees C preservation groups as compared to the 4 degrees C group (p less than 0.0001). This study shows that significant improvement in islet yield and function occurs when the rat pancreas was preserved at 7-10 degrees C rather than at 4 degrees C.

Animals↗

PubMed1991

Cardiac mechanical restitution in active and hibernating Richardson's ground squirrel.

The cardiac mechanical restitution was compared in papillary muscles between the active and the hibernating Richardson's ground squirrels at 0.1, 2.8, and 5 mM external Ca2+ concentration [( Ca2+]o). The amplitude of the restitution was significantly higher in hibernating animals between 37 and 7 degrees C at all [Ca2+]o. The first postrest contraction (F1) was highest at 20 degrees C and lower at 37 and 7 degrees C in both groups. The pause duration for maximum F1 was 30 s in active but 10 s in hibernating animals at 37 degrees C and increased to 100 s in both groups at 7 degrees C. The postrest potentiation was eliminated by 10(-6) M ryanodine at 20 degrees C in both groups, and this inhibitory effect was more pronounced in the hibernating group. Together, our results suggest that the activator Ca2+ for excitation-contraction coupling is mainly derived from the sarcoplasmic reticulum (SR) pool in both active and hibernating ground squirrel, and the dependence on SR Ca2+ release via ryanodine-sensitive Ca2+ channels is more marked in the hibernating state. Furthermore, there is no significant difference in sensitivity of the cardiac mechanical restitution to [Ca2+]o between the active and the hibernating condition.

Activity Cycles↗

PubMed1991

Plasma glucagon, glucose, and free fatty acid concentrations and secretion during prolonged hypothermia in rats.

Impairment of metabolic substrate mobilization and utilization may be a factor limiting survival in hypothermia. Using a newly developed technique for maintaining stable low body temperature (Tb), substrate profiles and their regulation by glucagon were examined in hypothermic rats (Tb 19 +/- 0.3 degrees C) over 20 h. During cooling, plasma glucagon, glucose, and free fatty acid (FFA) concentrations increased significantly (536 +/- 55 pg/ml, 304 +/- 26 mg/100 ml, and 844 +/- 81 mueq/l, respectively). Plasma glucagon and glucose concentrations continued to increase up to 8 h (peaks 810 +/- 103 pg/ml and 451 +/- 33 mg/100 ml, respectively) and remained high throughout the rest of the hypothermic period. FFA concentrations decreased steadily during the hypothermic period. Exogenous glucagon (20 micrograms/kg) induced significant increases in plasma glucose (+129 +/- 31 mg/100 ml) and FFA concentrations (+351 mueq/l) at 2 h but had no effect at 15 h of hypothermia. In vitro evaluation of pancreatic alpha-cell function indicated that glucagon secretion is independent of temperature between 37 and 19 degrees C. Our data indicate that hypothermia is characterized by a disturbed substrate metabolism, which is likely due to an imbalance in pancreatic alpha- and beta-cell function and a time-dependent decrease in tissue sensitivity to glucagon. These deleterious changes may limit survival in hypothermia.

Animals↗

PubMed1991

Effects of ginsenoside Rb1 on central cholinergic metabolism.

Ginsenosides, the saponins of ginseng, are bioactive ingredients which exert many beneficial effects. One ginsenoside, Rb1, extracted from North American ginseng (Panax quinquefolium L.), partially prevents the memory deficits induced by a cholinergic agent (scopolamine) in rats. In vitro studies show that Rb1 has no effect on quinuclidinyl benzylate binding or on acetylcholinesterase activity, but facilitates the release of acetylcholine (ACh) from hippocampal slices. The increase in ACh release is associated with an increased uptake of choline into nerve endings; however, calcium influx is unaltered. The ability of Rb1 to prevent memory deficits may be related to facilitation of ACh metabolism in the central nervous system.

Acetylcholine↗

PubMed1990

Berberine-induced morphologic differentiation and down-regulation of c-Ki-ras2 protooncogene expression in human teratocarcinoma cells.

A pluripotent human teratocarcinoma cell clone, NT2/D1, which was derived from the Tera-2 cell line, was induced to differentiate into cells with neuronal cell morphology by treatment with berberine. As early as 1 day after a 24-h treatment of cells with berberine at a non-toxic dose of 0.1 mg/ml in culture medium, the cells started to show morphologic changes, developing into terminally differentiated neuronal cells with long, inter-connecting network-like cellular structures. This process is much faster as compared with that induced by treatment with retinoic acid (RA), which took at least several days to develop. Unlike RA, berberine could not induce murine teratocarcinoma cell line, F9, to differentiate into endodermal cells. It was also found that, although the NT2/D1 cell clone exhibited amplification and enhanced mRNA expression of c-Ki-ras2 gene as did the parent cell line, a marked down-regulation of c-Ki-ras2 mRNA expression was observed. However, there was no change in actin mRNA expression even after differentiation had occurred. Thus, morphologic differentiation of teratocarcinoma cells into neuronal cells is found to be associated with down-regulation of a protooncogene which plays some definite role in oncogenesis. The mechanism by which berberine induces differentiation in these cells needs further investigation.

Animals↗

PubMed1990

Improvement of cold tolerance by selective A1 adenosine receptor antagonists in rats.

Previously we have shown that the improvement of cold tolerance by theophylline is due to antagonism at adenosine receptors rather than inhibition of phosphodiesterase. Since theophylline is a nonselective adenosine receptor antagonist for both A1 and A2 receptors, the present study investigated the adenosine receptor subtype involved in theophylline's action. Acute systemic injection of selective A1 receptor antagonists (1,3-dialkyl-8-aryl or 1,3-dialkyl-8-cyclopentyl xanthine derivatives) significantly increased both the total and maximal heat production as well as cold tolerance. In contrast, injection of a relatively selective A2 receptor antagonist, 3,7-dimethyl-1-propargylxanthine (compound No. 19), failed to significantly alter the thermogenic response of the rat under cold exposure. Further, the relative effectiveness of these compounds in increasing total thermogenesis was positively correlated with their potency in blocking the A1 adenosine receptor (r = .52, p less than 0.01), but not in A2 adenosine receptor (r = .20, p less than 0.2). It is likely that the thermally beneficial effects of adenosine A1 antagonists are due to their attenuation of the inhibitory effects of endogenously released adenosine on lipolysis and glucose utilization, resulting in increased substrate mobilization and utilization for enhanced thermogenesis.

Animals↗