NGF mRNA is expressed by GABAergic but not cholinergic neurons in rat basal forebrain

Nerve growth factor (NGF) supports the survival and biosynthetic activities of basal forebrain cholinergic neurons and is expressed by neurons within lateral aspects of this system including the horizontal limb of the diagonal bands and magnocellular preoptic areas. In the present study, colormetric and isotopic in situ hybridization techniques were combined to identify the neurotransmitter phenotype of the NGF-producing cells in these two areas. Adult rat forebrain tissue was processed for the colocalization of mRNA for NGF with mRNA for either choline acetyltransferase, a cholinergic cell marker, or glutamic acid decarboxylase, a GABAergic cell marker. In both regions, many neurons were single-labeled for choline acetyltransferase mRNA, but cells containing both choline acetyltransferase and NGF mRNA were not detected. In these fields, virtually all NGF mRNA-positive neurons contained glutamic acid decarboxylase mRNA. The double-labeled cells comprised a subpopulation of GABAergic neurons; numerous cells labeled with glutamic acid decarboxylase cRNA alone were codistributed with the double-labeled neurons. These data demonstrate that in basal forebrain GABAergic neurons are the principal source of locally produced NGF.     

Repeated cold water swim produces delayed nociceptive responses, but not analgesia, for tonic pain in the rat

Earlier studies have demonstrated that cold water swim (CWS) produces stress-induced analgesia in tests of brief, phasic pain and produces a delayed nociceptive response (DNR) for more prolonged tonic pain. The present study reports the effect of repeated CWS on tonic pain, as measured by the formalin test. One group of rats was exposed to a 3.5-min swim in 2 degrees C water immediately prior to the formalin injection, to a 1.5-min swim at 50 min, and to another 1.5-min swim at 100 min postformalin injection. Compared to the no-swim control group, subjects which received repeated CWS had dramatically altered formalin pain responses. Formalin responses began just over 3 h postformalin injection, peaked at 4 h, and were still present at 5 h. Inspection of individual responses revealed a substantial degree of variability in the onset of responses, although the magnitude and duration of the formalin pain response remained at the same levels as those of control subjects. The lack of a decrease in the magnitude and duration of the delayed formalin responses indicates that repeated CWS does not produce analgesia for tonic pain. The period of stress, therefore, produces pain suppression but not loss of the mechanisms that subsequently underlie the pain. Earlier controls have ruled out peripheral mechanisms (such as retention of the formalin in the paw tissue). Rather, a memory mechanism appears to have been indicated and it is not lost, but persists until it can be manifested. Further research is needed to study the mechanisms responsible for the DNR.     

Effects of reserpine on ECL-cell ultrastructure and histamine compartmentalization in the rat stomach

The histamine-storing ECL cells in the stomach play a key role in the control of acid secretion. They contain granules, secretory vesicles and microvesicles, and sustained gastrin stimulation results in the additional formation of vacuoles and lipofuscin bodies. The cells are rich in the vesicle monoamine transporter type-2 (VMAT-2), which can be inhibited by reserpine. The present study examines the effect of reserpine on ECL-cell ultrastructure and histamine compartmentalization. Rats received reserpine and/or gastrin. Reserpine was given twice by the intraperitoneal route (25 mg/kg once daily). Gastrin-17 was given by subcutaneous infusion (5 nmol/kg/h), starting at the time of the first reserpine injection and continuing for 4 days when the rats were killed. At this stage, histamine in the oxyntic mucosa was unaffected by reserpine but elevated by gastrin. Immunocytochemical analysis (confocal microscopy) showed ECL-cell histamine in control and gastrin-treated rats to be localized in cytoplasmic organelles (e.g., secretory vesicles). After treatment with reserpine alone or reserpine+gastrin, ECL-cell histamine occurred mainly in the cytosol. Planimetric analysis (electron microscopy) of ECL cells showed reserpine to increase the number, size and volume density of the granules and to reduce the size and volume density of the secretory vesicles. Gastrin reduced the number and volume density of granules and secretory vesicles, increased the number and volume density of microvesicles and caused vacuoles and lipofuscin bodies to appear. Reserpine+gastrin increased the number, volume density and size of the granules. Reserpine prevented the effects of gastrin on secretory vesicles, vacuoles and microvesicles, but did not prevent the development of lipofuscin. Our findings are in line with the views: (1) that preformed cytosolic histamine is taken up by granules/secretory vesicles via VMAT-2, that histamine is instrumental in the transformation of granules into secretory vesicles and in their consequent enlargement and (2) that vacuoles are formed by the fusion of large secretory vesicles.     

Metabolic compromise with systemic 3-nitropropionic acid produces striatal apoptosis in Sprague-Dawley rats but not in BALB/c ByJ mice

Metabolic compromise with systemic 3-nitropropionic acid (3-NP) results in the degeneration of striatal cells, mimicking the pathology of Huntington's disease (HD). Here we show that 10-week- and 8-month-old BALB/c ByJ mice show an unexpected striatal resilience to single and multiple systemic injections of 3-NP, while Sprague-Dawley rats are vulnerable, albeit in a variable manner. Identification of lesions was made by staining of DNA fragmentation with terminal deoxytransferase-mediated dUTP-biotin nick-end labeling (TUNEL) and hematoxylin/eosin, 1-10 days after injection. Quantitative imaging of histochemistry for succinate dehydrogenase (SDH) activity, the target of 3-NP inhibition, revealed that vulnerable rats reached maximal inhibition in brain at 1 day after 3-NP, whereas mice and resilient rats took 7 days to reach maximal inhibition. All groups of animals reached similar maximal decreases in SDH activity in striatum and cortex. Remarkably, only the fast decline in SDH activity seen in vulnerable rats was associated with TUNEL labeling. In addition, vulnerable rats developed a region within striatum where SDH activity was fully depleted and a similarly depleted region in CA1 hippocampus. While mice did not develop this region in striatum, some developed one in CA1. These regions of SDH depletion in both structures were associated with widespread TUNEL staining, with maximal labeling at 3 days after 3-NP. The existence of an animal strain resilient to 3-NP suggests that there are mediating factors involved in the preferential vulnerability of striatum to metabolic lesioning. The identification of these factors could provide strategies for therapeutic intervention in HD.     

Valyl-transfer RNA formation stimulated by monovalent cations and spermine in rat liver

The effects of hypothalamic disconnection on body temperature and hypothalamo-pituitary-adrenal activity following acute and repeated exposures to heat were studied. Intact male rats, or animals with complete posterior or anterior hypothalamic disconnection, were exposed to a temperature of 36 degrees C and a relative humidity of 35-45%. In the complete posterior and anterior hypothalamic disconnected rats the basal Tre was higher than that of the intact rats; the rise in Tre following heat exposure was lower in the operated rats than in the intact animals. All the experimental animals, except for those with anterior hypothalamic disconnection, showed a significant inhibition of corticosterone release on exposure to heat for 30 min, but no inhibition was observed in any of the disconnected rats when they were exposed to heat for 120 min. These results suggest that the main stimulus for ACTH release, during the first 30 min of heat exposure, is mediated by a neural input through the posteroir hypothalamus and this is followed by a nerural and/or humoral mechanism which enables the animals to increase their corticosterone secretion.     

Projections from the lateral, basal, and accessory basal nuclei of the amygdala to the hippocampal formation in rat

The amygdaloid complex and hippocampal formation mediate functions involving emotion and memory. To investigate the connections that regulate the interactions between these regions, we injected the anterograde tracer Phaseolus vulgaris-leucoagglutinin into various divisions of the lateral, basal, and accessory basal nuclei of the rat amygdala. The heaviest projection to the entorhinal cortex originates in the medial division of the lateral nucleus which innervates layer III of the ventral intermediate and dorsal intermediate subfields. In the basal nucleus, the heaviest projection arises in the parvicellular division and terminates in layer III of the amygdalo-entorhinal transitional subfield. In the accessory basal nucleus, the parvicellular division heavily innervates layer V of the ventral intermediate subfield. The most substantial projection to the hippocampus originates in the basal nucleus. The caudomedial portion of the parvicellular division projects heavily to the stratum oriens and stratum radiatum of CA3 and CA1. The accessory basal nucleus projects to the stratum lacunosum-moleculare of CA1. The subiculum receives a substantial input from the caudomedial parvicellular division. The parasubiculum receives dense projections from the caudal portion of the medial division of the lateral nucleus, the caudomedial parvicellular division of the basal nucleus, and the parvicellular division of the accessory basal nucleus. Our data show that select nuclear divisions of the amygdala project to the entorhinal cortex, hippocampus, subiculum, and parasubiculum in segregated rather than overlapping terminal fields. These data suggest that the amygdaloid complex is in a position to modulate different stages of information processing within the hippocampal formation.     

Aging stimulates fatty acid oxidation in rat colonocytes but does not influence the response to dietary fiber

Metabolism was studied in colonocytes isolated from young (4 mo) and aged (24 mo) Fischer 344 rats. Animals were fed fiber-free, low-fiber (5% cellulose), or high-fiber (oat bran or NIH 31 stock) diets. Colonocytes isolated from aged animals oxidized both short- and long-chain fatty acids at significantly higher rates than did colonocytes isolated from young animals. No differences between the young and aged were noted for the oxidation to CO2 of glucose and glutamine or for flux of glucose through glycolysis. Net adenosine triphosphate (ATP) production by colonocytes was calculated to be 20% higher for the aged than for the young, although the relative contribution of substrates to net ATP production from exogenous substrates was similar for the young and aged (45-50% from butyrate, 20-25% from glucose, and 30% from other substrates including acetate, propionate, palmitate, and glutamine). Substrate oxidation was generally higher in colonocytes from the oat bran (17% total dietary fiber, highly soluble fiber) versus fiber-free diet.     

LSD produces place preference and flavor avoidance but does not produce flavor aversion in rats.

The hedonic properties of lysergic acid diethylamide (LSD) were assessed using the place conditioning, taste reactivity, and taste avoidance tests. LSD produced a conditioned place preference, but only at the highest dose tested (0.2 mg/kg). A single preexposure to the conditioning chamber (latent inhibition) prevented the establishment of a place preference. When paired with sucrose, doses of 0.05 to 0.2 mg/kg of LSD produced taste avoidance, but no dose of LSD produced an aversion to the taste as assessed by the taste reactivity test. These results suggest that LSD, like other rewarding drugs, produces taste avoidance by a mechanism other than that produced by emetic drugs. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Functional expression of the apical Na+-dependent bile acid transporter in large but not small rat cholangiocytes

BACKGROUND & AIMS: Bile acids interact with cholangiocytes, resulting in cholangiocyte proliferation and increases in ductal bile secretion in large but not small cholangiocytes. It was proposed that for bile acids to exert these effects on cholangiocytes, a specific uptake mechanism must be present in cholangiocytes. The aim of this study was to show the expression of a bile acid transporter in cholangiocytes. METHODS: Small and large cholangiocytes or intrahepatic bile duct units (IBDUs) were isolated from normal rats, and gene expression for the apical Na+-dependent bile acid transporter (ABAT) and the 14-kilodalton ileal cytosolic binding protein (IBABP) was assessed by ribonuclease-protection assays. Tissue and subcellular distribution of bile acid transporters was also studied. [14C]-Taurocholate uptake into cholangiocytes was determined. RESULTS: Both ABAT and IBABP messenger RNAs were detected in large but not small cholangiocytes. By immunohistochemistry, ABAT was present in large but not small cholangiocytes. Immunofluorescence showed ABAT to be present in the apical membrane of large IBDUs. A Na+-dependent saturable uptake of taurocholate was present in large but not small cholangiocytes. CONCLUSIONS: These proteins may mediate bile acid uptake from the duct lumen in large ducts, resulting in modification of canalicular bile secretion and modulation of ductal bile secretion and growth.     

Effects of anaesthetics and trauma on stimulated gastric acid secretion in chronic fistula dogs

1. The present study was undertaken to determine whether various anaesthetic agents affect canine gastric acid secretion independently of other experimental variables. 2. Acid secretory output was determined in dogs with chronic fistulae, by administering sedating doses of anaesthetics commonly used for studying gastric secretory mechanisms in laboratory animals. 3. The anaesthetic agents inhibited gastric acid secretion. As the inhibitory effect of the mixture of anaesthetics was pronounced, an attempt was made to study the effect of each individual anaesthetic agent separately. 4. Acetopromazine was given to sedate dogs. Although it has a long duration of action, it only had a transient inhibitory action on gastric acid secretion of 15-30 min duration. Moreover the drug reduced pentagastrin-stimulated secretion, but had no effect on histamine-stimulated secretion. 5. Thiopentone sodium given with acetopromazine produced a mild inhibitory effect on histamine-stimulated secretion for 45 min, but produced a more pronounced and sustained inhibitory effect on pentagastrin-stimulated secretion. 6. Trilene significantly inhibited both histamine- and pentagastrin-stimulated secretion. The effect on the latter was more pronounced and sustained. 7. Trauma had no significant effect on histamine-stimulated secretion, but showed a slight inhibitory effect on pentagastrin-stimulated secretion. 8. Experiments to study gastric secretory mechanisms and antisecretory drugs should take account of the potential inhibitory effects of anaesthetics. Where possible, studies in conscious dogs with gastric fistulae are preferable to experiments on anaesthetized animals.     

Amyloid beta-peptide and its fragments induce acetylcholine release in in vitro basal forebrain tissue slices of rat brain, but do not affect the choline uptake

OBJECTIVE: Although stressor uncontrollability has been shown to suppress immune responses in animals and for human subjects, the results have been inconsistent. We reanalyzed results of our previous study regarding stress-related immune deviation in man, to establish whether perceived uncontrollability of an acute stressor acts as a co-determinant in the observed changes in immunological parameters. METHOD: Three types of cognitive reactions to an acute interpersonal stressor were assessed: "motivation," "uncontrollability," and "guiltiness." Stress-induced changes in the number of several types of immune cells in peripheral blood and proliferative responses of lymphocytes to antigens and mitogens were assessed. RESULTS: In comparison with control subjects and with subjects perceiving high control over the experimental stress situation, the subject perceiving low control showed a stressor-induced decrease in the number of T helper cells. Reversely, subjects perceiving high control showed an increase in the number of B cells as opposed to the other two groups. The effects of perceived uncontrollability could not be accounted for by mood changes, but they were related to previously experienced life stress. CONCLUSIONS: Perceived uncontrollability of an acute stressor can have immuno-modulating effects over and above those of the stressor per se.     

Evidence for anatomical but not functional asymmetry in the hemidecorticate rat.

Studied 51 male Long-Evans rats with unilateral ablation of the left or right cerebral neocortex or left or right hippocampus using a battery of tests of spatial orientation, motor coordination, and social behavior, including Morris water task, radial arm maze, feeding, narrow beam traversing, puzzle latches, hoarding, grooming, nest building, running wheel activity, male–male interaction, and shock-induced aggression. Comparison of the brains of operated and control Ss confirmed previous suggestions that the right hemisphere of the rat is bigger and may have different connections than the left hemisphere. Despite the morphological asymmetries, comparison of the behavior of Ss with right and left hemidecortication and right and left hippocampal lesions failed to show a single instance of functional asymmetry in the rat brain. These behavioral results contrast with previous reports of functional asymmetry in the control of activity, orientation, and rotation. (32 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Gastric secretion as a function of acute and chronic stress in the gastric fistula rat.

Conducted 2 experiments using a total of 54 male Long-Evans rats chronically implanted with gastric cannulas. In Exp I Ss exposed to signaled and unsignaled grid shock secreted more gastric acid after shock stress (chronic stress) for 8 days compared to the 1st 12 hrs of shock stress (acute stress). However, Exp II indicated that the higher gastric acid values under chronic stress were not significantly greater than prestress baseline values. Results are interpreted to reflect an inhibition of gastric acid secretion as a function of acute stress. During chronic stress this inhibition was followed by an habituation of gastric secretory processes which was observed as a return of secretion volume to baseline levels. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Tau glycation is involved in aggregation of the protein but not in the formation of filaments

A tau peptide, peptide 2R, with capacity for self assembly into filaments was used as a model to test the role of glycation on tau assembly or aggregation. Our results indicate that glycation of that peptide facilitates dimer formation but not assembly into filaments. However, glycation of tau results in the bundling of the tau filaments formed by glycosaminoglycan-induced polymerisation. These results suggest a role of glycation in the formation of covalent links among pre-formed filaments but not in the assembly of those filaments.     

Dietary alpha-linolenic acid increases the biosynthesis of the choline glycerophospholipids from [14C]CDPcholine in rat liver and kidney but not in brain

The effect of feeding rats for 30 days with diets containing high levels of linoleic acid (sunflower oil, SO) or alpha-linolenic acid (perilla oil, PO) was studied in the liver, kidney and brain. The PO group showed a higher labeling of choline glycerophospholipids (CGP) in liver and kidney but no difference with the SO group in ethanolamine glycerophospholipids (EGP) labeling. The brain displayed the lowest incorporation of both precursors and no difference between the two diets. Analyses of brain CGP and EGP fatty acid composition showed that in the PO group the ratio n-6/n-3 was lower than in the SO group, mainly as a consequence of lower levels of n-6 fatty acids. The mole % of docosahexaenoate (DHA) in these lipids was the same for both groups and only triacylglycerols (TAG) displayed a higher DHA. Therefore, at least in the brain, the magnitude of fatty acid changes observed in CGP and EGP for the PO group does not affect the uptake/incorporation of the precursors into phospholipids.     

Estrogen regulates galanin but not tyrosine hydroxylase gene expression in the rat locus ceruleus

The neuropeptide galanin (GAL) is coexpressed by the majority of noradrenergic neurons in the rat locus ceruleus (LC) and may function as an inhibitory modulator of noradrenergic transmission. Because estrogen has been shown to induce GAL expression in other brain regions and modulate noradrenergic transmission, we used in situ hybridization histochemistry to assess the effects of chronic estrogen treatment on GAL and tyrosine hydroxylase (TH) gene expression in the LC of ovariectomized female rats. We found that GAL mRNA levels were significantly elevated in rats implanted with a Silastic capsule containing estradiol compared to sham-implanted controls. Both the average optical density (P < or = 0.05) and the labelling area (P < or = 0.007) differed significantly between the groups. In contrast, TH gene expression measured in alternate brain sections did not differ between the groups. If GAL functions as an inhibitory modulator of noradrenergic transmission as postulated, these findings suggest that chronic estrogen treatment could reduce the noradrenergic tone of the brain in the absence of significant alterations in TH expression by enhancing the level of cosecreted GAL.     

Spontaneous recovery but not reinstatement of the extinguished conditioned eyeblink response in the rat.

Reinstatement—the return of an extinguished conditioned response (CR) after reexposure to the unconditioned stimulus (US)—and spontaneous recovery—the return of an extinguished CR with the passage of time—are 2 of 4 well-established phenomena that demonstrate that extinction does not erase the conditioned stimulus (CS)–US association. However, reinstatement of extinguished eyeblink CRs has never been demonstrated, and spontaneous recovery of extinguished eyeblink CRs has not been systematically demonstrated in rodent eyeblink conditioning. In Experiment 1, US reexposure was administered 24 hr prior to a reinstatement test. In Experiment 2, US reexposure was administered 5 min prior to a reinstatement test. In Experiment 3, a long, discrete cue (a houselight), present in all phases of training and testing, served as a context within which each trial occurred to maximize context processing, which in other preparations has been shown to be required for reinstatement. In Experiment 4, an additional group was included that received footshock exposure, rather than US reexposure, between extinction and test, and contextual freezing was measured prior to test. Spontaneous recovery was robust in Experiments 3 and 4. In Experiment 4, context freezing was strong in a group given footshock exposure but not in a group given eye shock US reexposure. There was no reinstatement observed in any experiment. With stimulus conditions that produce eyeblink conditioning and research designs that produce reinstatement in other forms of classical conditioning, we observed spontaneous recovery but not reinstatement of extinguished eyeblink CRs. This suggests that reinstatement, but not spontaneous recovery, is a preparation- or substrate-dependent phenomenon. (PsycINFO Database Record (c) 2011 APA, all rights reserved)