Biomedical subjects

M J Glass

Publications and source records attributed to M J Glass.

At least 19 recordsLinked to original sources

PubMed2006

Subcellular localization of nicotinamide adenine dinucleotide phosphate oxidase subunits in neurons and astroglia of the rat medial nucleus tractus solitarius: relationship with tyrosine hydroxylase immunoreactive neurons.

Superoxide produced by the enzyme nicotinamide adenine dinucleotide phosphate (NADPH) oxidase mediates crucial intracellular signaling cascades in the medial nucleus of the solitary tract (mNTS), a brain region populated by catecholaminergic neurons, as well as astroglia that play an important role in autonomic function. The mechanisms mediating NADPH oxidase (phagocyte oxidase) activity in the neural regulation of cardiovascular processes are incompletely understood, however the subcellular localization of superoxide produced by the enzyme is likely to be an important regulatory factor. We used immunogold electron microscopy to determine the phenotypic and subcellular localization of the NADPH oxidase subunits p47(phox), gp91(phox,) and p22(phox) in the mNTS in rats. The mNTS contains a large population of neurons that synthesize catecholamines. Significantly, catecholaminergic signaling can be modulated by redox reactions. Therefore, the relationship of NADPH oxidase subunit labeled neurons or glia with respect to catecholaminergic neurons was also determined by dual labeling for the superoxide producing enzyme and tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine biosynthesis. In the mNTS, NADPH oxidase subunits were present primarily in somatodendritic processes and astrocytes, some of which also contained TH, or were contacted by TH-labeled axons, respectively. Immunogold quantification of NADPH oxidase subunit localization showed that p47(phox) and gp91(phox) were present on the surface membrane, as well as vesicular organelles characteristic of calcium storing smooth endoplasmic reticula in dendritic and astroglial processes. These results indicate that NADPH oxidase assembly and consequent superoxide formation are likely to occur near the plasmalemma, as well as on vesicular organelles associated with intracellular calcium storage within mNTS neurons and glia. Thus, NADPH oxidase-derived superoxide may participate in intracellular signaling pathways linked to calcium regulation in diverse mNTS cell types. Moreover, NADPH oxidase-derived superoxide in neurons and glia may directly or indirectly modulate catecholaminergic neuron activity in the mNTS.

Animals↗

PubMed2005

Angiotensin II AT-1A receptor immunolabeling in rat medial nucleus tractus solitarius neurons: subcellular targeting and relationships with catecholamines.

The angiotensin II AT-1A receptor (AT-1A) is the major mediator of the hypertensive actions of angiotensin II (ANG II) in the medial nucleus of the solitary tract (mNTS). The localization of the AT-1A receptor at surface or intracellular sites is an important determinant of its signaling properties, including intercellular or intracrine communication. However, the spatial localization of this protein, particularly within small distal or intermediate size dendrites of mNTS neurons, is unknown. Within the mNTS, ANG II and catecholamines interact in the regulation of autonomic function; however, it is unknown if AT-1A receptors are present at functional sites in catecholamine containing dendrites, or are contacted by catecholamine containing axon terminals. We compared surface and intracellular distributions of the AT-1A receptor in dendritic processes from the mNTS using immunogold electron microscopy in conjunction with immunoperoxidase labeling for tyrosine hydroxylase (TH) and morphometric analysis. Collapsed across all AT-1A-labeled dendritic profiles, immunogold labeling was more frequent in intracellular sites as compared with the plasma membrane. Small (<0.6 microm) dendritic profiles contained a higher ratio of particles associated with the surface membrane when compared with larger profiles. Approximately 27% of all AT-1A receptor-labeled dendritic profiles also contained labeling for TH. Approximately 12% of dendritic profiles single labeled for the AT-1A receptor were contacted by TH containing axons or axon terminals. The present results provide the first quantitative demonstration of select plasmalemmal and intracellular localizations of AT-1A receptors in dendritic processes of mNTS neurons, including those containing TH, or contacted by catecholaminergic axon terminals. These results suggest that AT-1A receptors are positioned for modulation of catecholamine signaling in the mNTS.

Animals↗

PubMed2001

Subcellular localization of alpha-2A-adrenergic receptors in the rat medial nucleus tractus solitarius: regional targeting and relationship with catecholamine neurons.

alpha-2A-adrenergic receptor (alpha2A-AR) agonists modulate diverse autonomic functions. These actions are believed to involve functionally specialized, second-order neurons in catecholamine-containing portions of the medial nucleus tractus solitarius (mNTS) at both intermediate (NTSi) and caudal (NTSc) levels. However, the cellular mechanisms subserving alpha2A-AR-mediated actions within the mNTS have yet to be established. Immunocytochemistry was employed to examine the subcellular distribution of alpha2A-AR in both the intermediate and caudal mNTS and its association with cells containing the catecholamine-synthesizing enzyme tyrosine hydroxylase (TH). Quantitative regional comparison using immunogold showed that this receptor was distributed differentially to dendrites (NTSi, 46%; NTSc, 31%) and glia (NTSi, 29%; NTSc, 48%) at different levels of the NTS. Somata, axons, and terminals less frequently contained alpha2A-AR. The subcellular distribution of alpha2A-AR relative to catecholaminergic neurons also was similar within both subregions. Approximately 50% of alpha2A-AR-labeled somata also contained TH. In somatic profiles, alpha2A-AR labeling was often found in the cytosol and in association with endoplasmic reticulum and Golgi complexes, sites of receptor synthesis and trafficking. Approximately 20% of alpha2A-AR-immunoreactive dendrites also contained TH, where the receptor was often found on extrasynaptic portions of the plasma membrane near unlabeled terminals, some of which made symmetric contacts. However, TH-labeled terminals and dendrites usually were detected in the neuropil at a short distance (<10 microm) from alpha2A-AR-labeled neurons. alpha2A-AR-labeled glia frequently apposed unlabeled dendrites and terminals and were often located near TH-immunoreactive dendrites. These results indicate that, within the mNTS, alpha2A-AR is involved in a variety of autonomic processes, including postsynaptic modulation of mostly noncatecholaminergic dendrites, as well as influencing glia functions.

Animals↗

PubMed2001

Feeding inhibition by urocortin in the rat hypothalamic paraventricular nucleus.

Ventricular administration of urocortin (UCN) inhibits feeding, but specific site(s) of UCN action are unknown. In the current studies we examined the effect of UCN in the hypothalamic paraventricular nucleus (PVN) on feeding. We tested UCN administered into the PVN in several paradigms: deprivation-induced, nocturnal, and neuropeptide Y (NPY)-induced feeding. We compared the effect of equimolar doses of UCN and corticotrophin releasing hormone (CRH) on NPY-induced and nocturnal feeding, determined whether UCN in the PVN produced a conditioned taste aversion (CTA) and induced changes in c-Fos immunoreactivity (c-Fos-ir) after UCN and NPY administration in the PVN. UCN in the PVN significantly decreased NPY and nocturnal and deprivation-induced feeding at doses of 1, 10, and 100 pmol, respectively. UCN anorectic effects lasted longer than those attributed to CRH. Ten and thirty picomoles UCN did not induce a CTA, whereas 100 pmol UCN produced a CTA. UCN (100 pmol) in the PVN neither increased c-Fos-ir in any brain region assayed nor altered c-Fos-ir patterns resulting from PVN NPY administration. These data suggest the hypothalamic PVN as a site of UCN action.

Animals↗

PubMed2001

Naloxone's effect on meal microstructure of sucrose and cornstarch diets.

The opioid receptor antagonist naloxone decreases consumption of high-sucrose diets but does not reduce cornstarch diet intake in energy-restricted rats. Sucrose-fed rats eat at a much higher rate, consuming more food than cornstarch-fed rats. We examined meal microstructure using an automated weighing system in food-restricted rats eating either a high-sucrose or high-cornstarch diet. Sucrose-fed rats exhibited a higher rate of eating during their first meal compared with cornstarch-fed rats (0.34 vs. 0.20 g/min, respectively). However, naloxone did not reduce eating rate in either group. Naloxone decreased the size of the first meal in both diet groups by shortening the length of the meal. Naloxone's anorectic effect was more potent in the sucrose-fed rats. These results indicate that naloxone's heightened anorectic effect on sucrose diet consumption is not "rate dependent." Naloxone's anorectic actions may be modulated by two conditions, the sensory properties of food and the energy state of the animal. Thus the elevated anorectic potency of naloxone in energy-restricted sucrose-fed rats may reflect actions on neural systems that mediate orosensory and/or postingestive signals.

Animals↗

PubMed2000

Regional effect of naltrexone in the nucleus of the solitary tract in blockade of NPY-induced feeding.

Naltrexone (NLTX) in the nucleus of the solitary tract (NTS) decreases feeding induced by neuropeptide Y (NPY) in the paraventricular nucleus (PVN). We sought to determine the NTS region most sensitive to NLTX blockade of PVN NPY-induced feeding. Male Sprague-Dawley rats were fitted with two cannulas; one in the PVN and one in a hindbrain region: caudal, medial, or rostral NTS or 1 mm outside the NTS. Animals received NLTX (0, 1, 3, 10, and 30 microg in 0.3 microl) into the hindbrain region just prior to PVN NPY (0.5 microg, 0.3 microl) or artificial cerebrospinal fluid (0.3 microl). Food intake was measured at 2 h following injection. PVN NPY stimulated feeding, and NLTX in the medial NTS significantly decreased NPY-induced feeding at 2 h, whereas administration of NLTX in the other hindbrain regions did not significantly influence PVN NPY induced feeding. These data suggest that opioid receptors in the medial NTS are most responsive to feeding signals originating in the PVN after NPY stimulation.

Animals↗

PubMed2000

Naltrexone administered to central nucleus of amygdala or PVN: neural dissociation of diet and energy.

There is evidence that opioids may affect food consumption through mechanisms as diverse as reward or energy metabolism. However, these hypotheses are derived from studies employing peripheral or, more rarely, intracerebroventricular administration of drugs. Opioid receptors have a wide distribution in the central nervous system and include a number of regions implicated in food intake such as the hypothalamic paraventricular nucleus (PVN) and the central nucleus of the amygdala (ACe). It is not known whether local opioid receptor blockade in either of these regions will produce similar effects on food intake. To examine this issue, a chronic cannula was aimed at either the PVN or ACe of rats that were fed a choice of a high-fat and high-carbohydrate diet, which allows for the measurement of both preference and total energy consumption. Naltrexone influenced preferred and nonpreferred food consumption, depending on the site of administration. Consumption of both preferred and nonpreferred diets was suppressed after PVN naltrexone administration, whereas only preferred diet intake was reduced after ACe injection of naltrexone. The present evidence indicates that direct stimulation of different brain regions with naltrexone may be associated with diverse effects on diet selection, which may be accounted for by manipulation of specific functional neural circuitry.

Amygdala↗

PubMed1999

The effect of naloxone on food-motivated behavior in the obese Zucker rat.

We assessed differences in food reinforced behavior between obese and lean Zucker rats with a progressive ratio schedule 3 (PR3) in which a subject emitted three additional lever-presses each time a reinforcer was delivered. The number of responses required for a reinforcer eventually exceeded its value, termed the "break point", a sensitive measure of food motivated behavior. Break points were higher in obese rats than lean controls for grain pellets (27.5 versus 9.5, P = 0.01) but not for sweet pellets (51.6 versus 38.5, P = 0.31). We determined if naloxone (0.01-3.0 mg/kg, SC), which reduces free food intake in obese Zucker rats, affects food motivated behavior in obese Zuckers and lean controls. Naloxone reduced break points in both obese and lean rats to a similar extent when working for either grain pellets or sweet pellets. Under free-access feeding conditions, naloxone again decreased pellet intake similarly in the obese and lean Zucker rats. Naloxone appeared to decrease free-access pellet consumption to a greater extent than break point in both groups. These results show that (1) obese rats exhibit higher levels of performance for food than lean rats only when working for the less valued grain pellet, (2) naloxone reduces both break points and free-access pellet consumption independent of genotype, and (3) naloxone appears to decrease food more effectively in rats given free access to food than in rats working for food.

Animals↗

PubMed1999

Opioids and food intake: distributed functional neural pathways?

Agonists of the mu, delta, kappa and ORL(1)opioid receptors increase food intake while opioid receptor blockade decreases food intake. The majority of the collected data related to opioids and feeding has led to the speculation that opioids are involved in meal maintenance and orosensory reward; however, some data suggest that opioids may impact feeding associated with energy needs. Based on the wide distribution of CNS opioid receptors and the presence of other neuropeptides in the vicinity of opioidergic pathways, it seems likely that opioids affect multiple feeding systems. For example, opioids in the hindbrain might be involved in both sensory and metabolic aspects of food intake, those in the amygdala in processing of 'emotional' properties of foods, and those in the hypothalamus in energy needs. In this review we present data which support functional diversity of opioids in feeding behavior.

Animals↗

PubMed1999

Role of lipid type on morphine-stimulated diet selection in rats.

Administration of morphine is said to increase fat consumption among rats allowed to self-select nutrients. However, fats represent a diverse group of molecules, differing in metabolic and sensory properties. Despite this, lipid has yet to be manipulated as a variable in drug-stimulated nutrient selection studies. To determine whether lipid source can impact daily and morphine-stimulated (1, 3, and 10 mg/kg) diet intake, rats were provided with a choice between a high-fat and high-carbohydrate diet in three regimens in which the source of fat was varied between vegetable shortening, lard, or corn oil. Daily and morphine-stimulated diet selections were determined under all conditions. Under daily feeding conditions, rats ate more of the high-lipid diet compared with the high-carbohydrate diet when vegetable shortening or lard was the main lipid alternative, but lipid and carbohydrate intake did not differ when corn oil was the main lipid alternative. When rats were stimulated with morphine, the percentage of lipid increased relative to baseline intake only when the lipid diets were the preferred alternatives (i.e., vegetable shortening or lard). When preference between lipid and carbohydrate diets was neutral (i.e., corn oil condition), morphine did not enhance lipid consumption. These results indicate that morphine increases consumption of total energy or preferred diets and not lipid per se.

Animals↗

PubMed1997

Role of carbohydrate type on diet selection in neuropeptide Y-stimulated rats.

We tested whether carbohydrate source (corn starch, sucrose, Polycose) influences the choice between a high-fat and high-carbohydrate diet in spontaneously feeding rats and in rats stimulated to eat by neuropeptide Y (NPY) administration or food deprivation. Rats were tested under three diet options: 1) a high-fat diet versus a high-corn starch diet; 2) a high-fat diet versus a high-sucrose diet, and 3) a high-fat diet versus a high-Polycose diet. During daily and stimulated feeding rats ate more of the high-carbohydrate diet than the fat diet when the source of carbohydrate was sucrose or Polycose; however, when corn starch was provided as the carbohydrate source rats ate more of the high-fat diet. Food-deprived rats increased intake of both the high-fat and the high-carbohydrate diets, with the proportion of energy ingested from each of the diets resembling that noted during 3 days of spontaneous feeding. NPY-injected rats ate more of both the high-fat and high-carbohydrate diets during diet options 1 and 3, but not during option 2 when the high-sucrose and high-fat diets were offered concurrently. In that case, rats did not significantly increase their intake of the high-fat diet. Although carbohydrate source and NPY administration each influenced diet selection, altering the source of carbohydrate had a more marked effect.

Animals↗

PubMed1996

Potency of naloxone's anorectic effect in rats is dependent on diet preference.

Modulation of feeding behavior by neuropeptide Y (NPY) and opioids is well established, but the possibility that these neural influences provoke specific appetites, NPY for carbohydrate and opioids for fat, has also been considered. In other studies, intake of standard chow after NPY stimulation can be blocked by naloxone, indicating an interaction between these systems in the regulation of feeding. The present experiments examined the nature of NPY-opioid interactions in diet selection. Rats were administered NPY and naloxone concurrently, then chose between high-fat and high-carbohydrate diets. Subcutaneous administration of naloxone (0.01-0.3 mg/kg) potently reduced intake of the preferred diet, but not the nonpreferred diet. A similar pattern of selection was seen in a separate experiment where the same doses of naloxone were administered after 24-h food deprivation. These data support the idea that the opioid system mediates the "rewarding" aspects of feeding.

Animals↗

PubMed1995

General, mu and kappa opioid antagonists in the nucleus accumbens alter food intake under deprivation, glucoprivic and palatable conditions.

Ventricular microinjection studies found that whereas mu (beta-funaltrexamine, B-FNA), mu1 (naloxonazine) and kappa (nor-binaltorphamine, Nor-BNI) opioid receptor antagonists, but not delta antagonists, reduce deprivation-induced intake, kappa and mu, but not mu1 or delta antagonists reduce both 2-deoxy-D-glucose (2DG) hyperphagia and sucrose intake. Since opioid agonists stimulate spontaneous food intake in the accumbens, the present study examined whether administration of either naltrexone, B-FNA or Nor-BNI in the accumbens altered intake under deprivation (24 h), glucoprivic (2DG: 500 mg/kg, i.p.) or palatable sucrose (10%) conditions. Naloxonazine's effects in the accumbens were also evaluated for deprivation-induced intake. Deprivation-induced intake was significantly decreased over 4 h by naltrexone (5-20 micrograms, 44%), B-FNA (1-4 micrograms, 55%) and Nor-BNI (4 micrograms, 31%) but not naloxonazine (10 micrograms) in the accumbens. 2DG hyperphagia was significantly decreased by naltrexone (10-20 microgram, 79%), B-FNA (1-4 micrograms, 100%) and NOR-BNI (104 micrograms, 75%) in the accumbens. Sucrose intake was significantly decreased by naltrexone (50 micrograms, 27%) and B-FNA (1-4 micrograms, 37%), but not NOR-BNI in the accumbens. These data suggest that mu receptors, and particularly the mu2 binding site in the accumbens are responsible for the opioid modulation of these forms of intake in this nucleus, and that this control may be acting upon the amount of intake per se.

Animals↗

PubMed1995

Alterations in deprivation, glucoprivic and sucrose intake following general, mu and kappa opioid antagonists in the hypothalamic paraventricular nucleus of rats.

While opioid agonists administered into the hypothalamic paraventricular nucleus increase food intake in rats, naloxone reduces deprivation-induced intake. Ventricular administration of either mu (beta-funaltrexamine) or kappa (nor-binaltorphamine) opioid antagonists reduces spontaneous, deprivation, glucoprivic and palatable intake. The present study assessed whether microinjections of either general, mu or kappa opioid antagonists into the paraventricular nucleus altered either deprivation (24 h) intake, 2-deoxy-D-glucose hyperphagia or sucrose intake in rats. Deprivation intake was significantly reduced by nor-binaltorphamine (5 micrograms, 68 nmol, 30-33%), beta-funaltrexamine (5 micrograms, 100 nmol, 26-29%) or naltrexone (10 micrograms, 260 nmol, 26%) in the paraventricular nucleus. 2-Deoxy-D-glucose hyperphagia was significantly reduced only after 2 h by naltrexone (10 micrograms, 260 nmol, 69%), norbinaltorphamine (20 micrograms, 272 nmol, 69%) or beta-funaltrexamine (20 micrograms, 400 nmol, 83%) in the paraventricular nucleus. Sucrose intake was significantly reduced by nor-binaltorphamine (5 micrograms, 68 nmol, 27-36%), naltrexone (5-10 micrograms, 130-260 nmol, 18-31%) and beta-funaltrexamine (5 micrograms, 100 nmol, 20%) in the paraventricular nucleus. These data indicate that general, mu and kappa opioid antagonists administered into the hypothalamic paraventricular nucleus produce similar patterns of effects upon different forms of food intake as did ventricular administration, implicating this nucleus as part of the circuitry underlying opioid mediation of ingestion.

Animals↗

PubMed1995

Analysis of central opioid receptor subtype antagonism of hypotonic and hypertonic saline intake in water-deprived rats.

Intake of either hypotonic or hypertonic saline solutions is modulated in part by the endogenous opioid system. Morphine and selective mu and delta opioid agonists increase saline intake, while general opioid antagonists reduce saline intake in rats. The present study evaluated whether intracerebroventricular administration of general (naltrexone) and selective mu (beta-funaltrexamine, 5-20 micrograms), mu, (naloxonazine, 50 micrograms), kappa (nor-binaltorphamine, 5-20 micrograms), delta (naltrindole, 20 micrograms), or delta 1 (DALCE, 40 micrograms) opioid receptor subtype antagonists altered water intake and either hypotonic (0.6%) or hypertonic (1.7%) saline intake in water-deprived (24 h) rats over a 3-h time course in a two-bottle choice test. Whereas peripheral naltrexone (0.5-2.5 mg/kg) significantly reduced water intake and hypertonic saline intake, central naltrexone (1-50 micrograms) significantly reduced water intake and hypotonic saline intake. Water intake was significantly reduced following mu and kappa receptor antagonism, but not following mu 1, delta, or delta 1 receptor antagonism. In contrast, neither hypotonic nor hypertonic saline intake was significantly altered by any selective antagonist. These data are discussed in terms of opioid receptor subtype control over saline intake relative to the animal's hydrational state and the roles of palatability and/or salt appetite.

Animals↗

PubMed1994

Central opioid receptor subtype mediation of isoproterenol-induced drinking in rats.

Opioid receptor subtype antagonists differentially alter different types of water intake such that mu2 receptors modulate deprivation-induced water intake, kappa receptors modulate hypertonic saline-induced water intake, and mu2, delta1 and kappa receptors modulate water intake following Angiotensin II (ANG II). Water intake stimulated by peripheral administration of the beta-adrenergic agonist, isoproterenol is attenuated by naloxone and is thought to be mediated by release of renin and production of ANG II. The present study examined whether systemic and i.c.v. administration of general opioid antagonists and central administration of specific opioid receptor subtype antagonists would selectively alter water intake following isoproterenol in rats. Both systemic (1 mg/kg s.c.) and central (1-20 micrograms) naltrexone reduced water intake induced by isoproterenol (25 micrograms/kg s.c.) over a 2-h period. The mu receptor antagonist, beta-funaltrexamine (B-FNA: 1-20 micrograms), but not the mu1 antagonist, naloxonazine (50 micrograms), dose-dependently reduced isoproterenol drinking. Both the kappa antagonist, nor-binaltorphamine (Nor-BNI, 5-20 micrograms) and the delta1 antagonist, [D-Ala2, Leu5, Cys6]-enkephalin (DALCE, 1-40 micrograms) also dose-dependently reduced isoproterenol drinking. These data implicate mu2, kappa and delta1 sites in the opioid modulation of isoproterenol drinking.

Animals↗

PubMed1993

Identification of the hepatitis A virus internal ribosome entry site: in vivo and in vitro analysis of bicistronic RNAs containing the HAV 5' noncoding region.

Hepatitis A virus (HAV), a RNA virus of positive polarity, contains a long 5' noncoding region (5'NCR) that lacks the characteristic m7GpppN cap group of most eukaryotic messages. By creating bicistronic constructs that contain the bacterial chloramphenicol acetyltransferase gene followed by the HAV 5'NCR and the luciferase gene we have demonstrated by assaying in vitro and in vivo that ribosome entry for translation initiation occurs via binding to sequences within the HAV 5'NCR. Using mutations created within this region we have identified that the HAV internal ribosome entry site (IRES) is located downstream of nucleotide 45 and including sequences up to nucleotide 734 of the HAV 5'NCR. Translation of a number of mutant constructs both in vitro in a rabbit reticulocyte lysate and in vivo by transfection of the cDNAs into BS-C-1 cells in the presence of the recombinant vaccinia virus, vTF7-3, gave similar results. However, a 4-nucleotide insertion at base 628 showed an increased activity over wild-type when transfected into BS-C-1 cells that was not seen in vitro. This increase in activity correlated with an increase in luciferase gene product as assayed by immunoprecipitations of [35S]methionine radiolabeled cells. Comparison of mono- and bicistronic RNAs that were synthesized with or without a m7GpppG cap group showed a competition for ribosome binding when translated in a rabbit reticulocyte lysate system. The presence of the cap group on the RNA 5'terminus of the RNA led to a greater ability of this RNA to translate than the RNA containing the HAV IRES.

Animals↗

PubMed1993

Identification of a trans-acting activity from liver that stimulates hepatitis A virus translation in vitro.

Hepatitis A virus (HAV), a picornavirus, is the causative agent of infectious hepatitis, generally a self-limiting disease of the liver. Recently, sequences within the 5' noncoding region that affect the translation of the viral genome have been identified using in vitro systems. In this report we demonstrate that extracts prepared from mouse liver cytoplasm specifically stimulate HAV RNA translation in a rabbit reticulocyte lysate in vitro. This activity appears to act specifically on HAV sequences and is not found in other mouse tissue and several cell lines of tissue culture origin.

Animals↗