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Thiamine transport in the central nervous system.

Total thiamine (free thiamine and thiamine phosphates) transport into the cerebrospinal fluid (CSF), brain, and choroid plexus and out of the CSF was measured in rabbits. In vivo, total thiamine transport into CSF, choroid plexus, and brain was saturable. At the normal plasma total thiamine concentration, less than 5% of total thiamine entry into CSF, choroid plexus, and brain was by simple diffusion. The relative turnovers of total thiamine in choroid plexus, whole brain, and CSF were 5, 2, and 14% per h, respectively, when measured by the penetration of 35S-labeled thiamine injected into blood. From the CSF, clearance of [35S]thiamine relative to mannitol was not saturable after the intraventricular injection of various concentrations of thiamine. However, a portion of the [35S]thiamine cleared from the CSF entered brain by a saturable mechanism. In vitro, choroid plexuses, isolated from rabbits and incubated in artificial CSF, accumulated [35S]thiamine against a concentration gradient by an active saturable process that did not depend on pyrophosphorylation of the [35S]thiamine. The [35S]thiamine accumulated within the choroid plexus in vitro was readily released. These results were interpreted as showing that the entry of total thiamine into the brain and CSF from blood is regulated by a saturable transport system, and that the locus of this system may be, in part, in the choroid plexus.

Animals↗

Release of vasopressin by angiotensin II.

To test the hypothesis that angiotensin II releases antidiuretic hormone (ADH) after injection into ventricular cerebrospinal fluid, conscious adult male Sprague-Dawley rats with a lateral cerebroventricular cannula received an intraventricular injection of 0, 10, 50, or 100 ng angiotensin II. Trunk blood was collected 90 seconds later for radioimmunoassay of ADH. Plasma ADH, pg/ml (mean plus or minus S. E.), for the four dose levels were 2.8 plus or minus 0.7, 9.6 plus or minus 2.5, 22.6 plus or minus 5.6 and 25.0 plus or minus 5.0, respectively. The increases produced by angiotensin were statistically significant (p smaller than 0.05). Plasma ADH of the 10 ng group was intermediate between control and the two highest angiotensin doses (p smaller than 0.05), suggesting a dose-response relationship. These data provide direct evidence that angiotensin releases ADH by central mechanisms.

Angiotensin II↗

beta-Endorphin and met-enkephalins: their distribution, modulation by estrogens and haloperidol, and role in neuroendocrine control.

A single dose of 0.5 micrograms beta-endorphin injected intraventricularly in unanesthetized male rats bearing chronic intraventricular and intrajugular cannulas led to a sevenfold stimulation of plasma prolactin (PRL) levels 10 to 20 min after injection of the peptide, while a dose of 2 micrograms of beta-endorphin led to a comparable stimulation of plasma growth hormone (GH) concentration. Met-Enkephalin was much less potent than beta-endorphin in stimulating PRL and GH release. Naloxone, a specific opiate antagonist, completely blocked the stimulation of GH and PRL release at the doses of 0.5 and 12.5 mg/kg, respectively. beta-Endorphin and Met-enkephalin from 41 discrete brain nuclei were measured by radioimmunoassay (RIA). beta-Endorphin was found in the hypothalamus medial preoptic nucleus, nucleus interstitialis striae terminalis (NIST), nucleus medialis thalami, and periaqueductal gray. Met-Enkephalin was found predominantly in the globus pallidus, NIST, medial preoptic nucleus, nucleus amygdaloideus centralis, and nucleus lateralis hypothalami. Treatment with both estrogens and haloperidol led to differential effects on Met-enkephalin content in various brain nuclei. Estrogen treatment increased Met-enkephalin levels in globus pallidus, NIST, medial and lateral preoptic nuclei, and periaqueductal gray, while a decrease of the Met-enkephalin content in the nucleus amygdaloideus centralis was found. Haloperidol treatment led to a stimulatory effect in striatum, medial and lateral preoptic nuclei, and interpeduncular nucleus.

Animals↗

On the possible role of brain protein synthesis in functional barbiturate tolerance.

Pentobarbital pellet implantation increased more than 200% the ED50 dose of pentobarbital required to induce loss of the righting reflex within 2 min of i.p. injection and increased the onset of barbital-induced sleep. Both tests of functional barbiturate tolerance were blocked by the intraventricular injection of cycloheximide. The effects of acute (45 mg/kg i.p.) and chronic (pellet implantation) pentobarbital treatment on the incorporation of 3/-lysine into the protein of various subcellular fractions of the cortex and subcortex were studied. In the subcortex, chronic pentobarbital treatment significantly stimulated protein synthesis 40-50% in the microsomal, soluble and mitochondrial fractions. Both acute and chronic pentobarbital treatments significantly increased (3H-lys)-protein accumulation in a fraction of synaptic plasma membranes derived from a population of nerve ending particles (NEP) enriched in gamma-aminobutyric acid (GABA). The possible significance of these data to pentobarbital tolerance and dependence development is discussed.

Animals↗

Ultrastructural localization and afferent sources of substance P in the rat parabrachial region.

The ultrastructural morphology and afferent sources of terminals containing substance P-like immunoreactivity were examined in the rat parabrachial region. In the first portion of the study, a polyclonal antiserum to substance P was localized in the ventrolateral parabrachial region using the peroxidase-antiperoxidase labeling technique combined with electron microscopy. The antiserum was tested for cross-reaction with substance P, physalaemin, substance K and neuromedins B, C and K. Cross-reactivity was most intense with substance P. However, substance K, neuromedin K and physalaemin also exhibited limited cross-reactions with the antiserum. In the ventrolateral parabrachial region of untreated adult animals, substance P-like immunoreactivity was localized in axon terminals containing numerous small (40-60 nm) clear vesicle and 1-3 large (90-120 nm) dense-core vesicles. At least 54% of the labeled terminals formed asymmetric synapses with unlabeled dendrites; and at least 30% of the recipient dendrites received more than one labeled axon terminal. In addition, the labeled terminals were associated less frequently with other unlabeled soma, axon terminals and blood vessels. In the second part of the study, we examined whether or not perikarya in various extrinsic regions contributed to the substance P-like immunoreactivity in axon terminals in the parabrachial region. Wheat-germ agglutinin conjugated horseradish peroxidase was injected unilaterally into the parabrachial region of adult rats two days prior to being killed and one day prior to intraventricular injection of colchicine (100 micrograms in 7.5 microliter saline) which enhanced the detection of immunoreactivity in perikarya. Sections were first processed by a tetramethylbenzidine reaction stabilized with cobalt-diaminobenzidine for demonstration of the transported peroxidase then were immunocytochemically labeled for substance P. Perikarya containing both the black granular retrograde labeling and brown peroxidase-immunoreactivity were found in the nuclei of the solitary tracts, the caudal ventrolateral reticular formation, the lateral dorsal tegmental nucleus and the paraventricular, dorsomedial and lateral hypothalamic nuclei. The projections were largely, but not exclusively, from perikarya located on the same side as the parabrachial injection. We conclude that substance P, or a closely related tachykinin, is a putative transmitter or modulator within a number of pathways to the parabrachial region and that these afferents act primarily through axodendritic synapses with intrinsic neurons.

Animals↗

Dual contribution theory of regulation of CSF HCO3 in respiratory acidosis.

Regulation of CSF HCO3-in respiratory acidosis was studied in light of the "dual contribution theory," which proposed that there were two sources for the CSF HCO3-increase: 1) HCO3-by diffusion from plasma and 2) HCO3-generated in the CNS and catalyzed by the local carbonic anhydrase (J. Appl. Physiol. 38: 504-512, 1975). In anesthetized dogs with an increase in Paco2 of 30 mmHg for 4 h the plasma HCO3 increased 2 meq/1 and CSF 6 meq/1. In combined respiratory and metabolic acidosis, plasma HCO3-did not increase but CSF HCO3-increased 6 meq/1. In combined acidosis and intraventricular injections of acetazolamide no increase in plasma or CSF HCO3-occurred. In combined respiratory acidosis and metabolic alkalosis and intraventricular acetazolamide, plasma HCO3-increased 15 meq/1 but CSF HCO3-increased 6 meq/1. Brain and CSF ammonia increased linearly and selectively with the increase in the relative contribution of CNS HCO3-increase. Therefore regulation of CSF HCO3-in respiratory acidosis depends on both components of the dual contribution theory, where each component can provide the total CSF HCO3-increase under appropriate experimental conditions. The control mechanism may be sensitive to changes in [H+] on the brain side of the blood-brain barrier.

Acid-Base Equilibrium↗

Role of brain histamine H1- and H2-receptors in neostigmine-induced hyperglycemia in rats.

We previously reported that when neostigmine, an inhibitor of acetylcholine esterase, was injected into the third cerebral ventricle, the concentration of hepatic venous plasma glucose was increased via central muscarinic receptors in anesthetized rats. To determine whether brain histamine receptors are involved in cholinergic system transmission with regard to central nervous system (CNS)-mediated glucoregulation, we examined the effects of the H1 receptor antagonist pyrilamine and the H2 receptor antagonist ranitidine on neostigmine-induced hyperglycemia in anesthetized rats. The injection of pyrilamine (5 x 10(-9)-5 x 10(-7) mol) into the third cerebral ventricle suppressed hyperglycemia induced by intraventricular injection of neostigmine (1 x 10(-9) mol) in a dose-dependent manner. Injection of ranitidine (5 x 10(-9)-5 x 10(-7) mol) into the third cerebral ventricle did not suppress the hyperglycemia induced by neostigmine, but enhanced it in a dose-dependent manner. These findings suggest that neostigmine-induced CNS-mediated hyperglycemia is transmitted by not only brain cholinergic muscarinic receptors but also in part by histamine H1 receptors.

Animals↗

Co-induction of p75(NTR) and the associated death executor NADE in degenerating hippocampal neurons after kainate-induced seizures in the rat.

Zinc induces in cultured cortical neurons both p75(NTR) and p75(NTR)-associated death executor (NADE), which together contribute to caspase-dependent neuronal apoptosis. Since zinc neurotoxicity may contribute to neuronal death following seizures, we examined whether p75(NTR) and NADE are co-induced also in rat hippocampal neurons degenerating after seizures. Staining of brain sections with a zinc-specific fluorescent dye (N-(6-methoxy-8-quinolyl)-p-carboxybenzoylsulphonamide) and acid fuchsin revealed zinc accumulation in degenerating neuronal cell bodies in CA1 and CA3 of hippocampus 24 h after kainate injection. Both anti-p75(NTR) and anti-NADE immunoreactivities appeared in zinc-accumulating/degenerating neurons in both areas. Intraventricular injection of CaEDTA, without altering the severity or time course of kainate-induced seizures, markedly attenuated the induction of p75(NTR)/NADE in hippocampus, which correlated with the decrease of caspase-3 activation and zinc accumulation/cell death. The present study has demonstrated that p75(NTR) and NADE are co-induced in neurons degenerating after kainate-induced seizures in rats, likely in a zinc-dependent manner.

Animals↗

Selective hippocampal lesions and behavior: effects of kainic acid lesions on performance of place and cue tasks.

Kainic acid (KA) lesions were used to study the effects of damage to the CA3 cell field and subiculum on performance of complex place and cue tasks. In Experiment 1, neuroanatomical techniques were employed to determine the selectivity of the lesions. In a within-subjects design, rats in Experiment 2 were trained before the operations to run on an eight-arm radial maze with procedures that involved two kinds of learning (place and cue) and two memory functions (reference memory and working memory). Interrupting the intrahippocampal circuit by damaging the CA3 cell field with KA had minimal effects on performance; injections into subiculum and complete aspiration lesions of hippocampus resulted in impairments on the place but not the cue task. Only intraventricular injections of KA affected performance on both tasks. These results fail to support either the cognitive map or the working memory theory of hippocampal function. The presence of distant damage beyond the immediate area of injection complicates interpretation of the results and may serve to limit the usefulness of KA as a neurotoxin in behavioral investigations.

Animals↗

Increased fucosylation of chick brain proteins following training: effects of cycloheximide.

When chicks are trained to avoid pecking a bead coated with methylanthranilate in a one-trial passive avoidance task there is an increase in fucose incorporation in vivo and in vitro in the right forebrain base of methylanthranilate (M)-trained compared to water (W)-trained chicks. The relation of this increase to de novo protein synthesis in vivo and in vitro has been examined. Cycloheximide (Cx), 1 mM, inhibited in vitro fucosylation of chick brain slices by 60% after 3 h. However, the training-related increase in in vitro fucosylation still persisted. When Cx was injected intraventricularly 10 min before training, the subsequent increase in in vitro fucosylation due to training was still apparent. When Cx was injected and [14C]leucine and [3H]fucose incorporation studied in vivo in M-trained and W-trained chicks, there was no increase in fucosylation due to training in the Cx-treated M-trained over the W-trained chicks. These results are taken to indicate that in vitro fucosylation and its increase subsequent to training is not protein synthesis-dependent, but that both in vivo and in vitro there are interactions between Cx and fucosylation steps that are independent of Cx's effects on protein synthesis.

Animals↗

Responses of rostral hypothalamic neurones to peripheral temperature and to amines.

1. Five-barrelled micropipettes have been used to record extracellularly the activity of neurones in the rostro-medial hypothalamus of methoxyflurane-anaesthetized cats, and to apply acetylcholine (ACh), noradrenaline (NA) and 5-hydroxytryptamine (5-HT) by micro-iontophoresis to the vicinity of each neurone encountered. Peripheral thermal stimulation was achieved by blowing warm (42 degrees C) and cold (4 degrees C) air in the face of the animal, and thermoresponsiveness was compared with amine responsiveness.2. One hundred and twenty-two neurones were obtained from ten cats. Eleven (9%) were warm-responsive and sixteen (13%) were cold-responsive. The rest did not respond to facial warming or cooling.3. No consistent relationship was observed between amine responses and responsiveness to facial temperature. Warm-responsive neurones were mainly depressed or unaffected by amines. Cool-responsive neurones were excited, depressed or unaffected by amines with the exception that no 5-HT excitations were seen. Thermoresponsive neurones were more likely to be amine depressed than non-thermoresponsive neurones.4. Six thermoresponsive neurones responded to peripheral temperature and to amines in a way which fitted the amine model of Myers (1971). Fifteen thermoresponsive neurones fitted the model of Bligh, Cottle & Maskrey (1971), according to the same criteria.5. The results lend little support to the amine model, as predicted from amine micro-injection and release studies in primates, but support more strongly the model of Bligh et al. (1971) which is based on intraventricular injections of amines in sheep, goats and rabbits. On the basis of the latter model, functional identification was possible in 63% of the thermoresponsive rostral hypothalamic neurones tested.

Acetylcholine↗

Blockade of brain M2 muscarinic receptors lowers blood pressure in spontaneously hypertensive rats.

Injections of the M2 muscarinic receptor antagonist 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP; 1.5-40 micrograms) into the cerebral ventricles of urethane-anesthetized rats caused a dose-related inhibition of the pressor response to intravenously injected physostigmine. A similar reduction was obtained with 1/80th the dose of methylatropine, but not with the selective M1 antagonist pirenzepine. Intraventricular injection of 4-DAMP (6.25-25 micrograms) caused a dose-related reduction in blood pressure in unanesthetized spontaneously hypertensive rats (SHR), but not in normotensive controls. Systolic pressure fell 42 +/- 6 mm Hg at the 25-micrograms dose. Pirenzepine did not lower blood pressure in SHR and inhibited the antihypertensive effect of 4-DAMP.

Animals↗

Role of brain catecholaminergic system in hypothalamic obesity in rats.

Distribution of labelled catecholamines (CA) was studied after intraventricular injection of 3H-norepinephrine (NE) and 3H-dihydrozyphenylalanine (DOPA) in several brain regions of ventromedial hypothalamic lesioned female rats. After intraventricular infection of 3H-NE a decrease in endogenous NE content and a marked increase in labelled NE concentration were observed in the preoptic-basal cortical area and lateral hypothalamus, while no change was found in these parameters in brain areas caudal to the lesion. The 3H-DOPA uptake of preoptic-basal cortical area and also the lateral hypothalamus was lower. No change was found in the uptake of 3H-DOPA in the corpus striatum. These results support the hypothesis that the decreased function of noradrenergic pathways in the medial forebrain bundle plays a role in the development of hypothalamic obesity without any concomitant functional changes in the striatal dopaminergic system.

Animals↗

Influence of ambient temperature on the thermoregulatory responses to 5-hydroxytryptamine, noradrenaline and acetylcholine injected into the lateral cerebral ventricles of sheep, goats and rabbits.

1. The influences of ambient temperature (T(a)) on the thermoregulatory effector activities and the body temperature (T(b)) of intraventricular injections into the sheep, goat and rabbit of 5-hydroxytryptamine (5-HT), noradrenaline (NA), acetylcholine (ACh), carbachol and eserine, have been interpreted in terms of a simple neuronal model of the pathways between thermosensors and thermoregulatory effectors.2. In all three species 5-HT in minimal doses caused a rise in respiratory frequency (RF) and a fall in T(b) at high T(a), and a reduction in EMG activity and a fall in T(b) at low T(a). These effects could be interpreted as those of an excitatory transmitter acting on the warm receptor-heat loss pathway.3. In all three species NA caused a reduction in RF and a rise in T(b) at high T(a), and a reduction in EMG activity and a fall in T(b) at low T(a). These effects are interpreted as those of an inhibitory transmitter acting both on the warm sensor-heat loss pathways and on the cold sensor-heat production pathway.4. The effects of ACh and the cholinomimetic substances carbachol and eserine are complex and more difficult to interpret. In small doses the effects on the sheep and goat are those of an excitatory transmitter on the cold sensor-heat production pathway. There was an increase in EMG activity and a rise in T(b) at low T(a), and a reduction in RF and a rise in T(b) at high T(a). At higher dose levels in the goat and at all dose levels in the rabbit these substances had the reverse effects which are attributed to a synaptic block due to the excess of the excitatory substance.5. The effects of ambient temperature and injected substances upon ear temperature are consistent with the predictions of the model if it is assumed (a) that at high and low ambient temperatures direct thermal effects on ear vessels dominate those of the sympathetic innervation, and (b) that the warm sensor influence is to lower peripheral vasomotor tone, and the cold sensor influence is to increase it.6. The conclusion reached is that when consideration is given to species differences in the thermoneutral ambient temperature and to the possibility that excitatory substances have reversed effects at high dose levels, the effects of 5-HT, NA and ACh in the control of body temperature are very similar in the sheep, goat and rabbit: 5-HT is excitatory on the heat loss pathway, ACh is excitatory on the heat production pathway and NA has an inhibitory influence on both pathways.

Acetylcholine↗

[Distribution, uptake and hydrolysis of cholesterol oleate (-1-14C) in brain tissue after its injection in the cerebral ventricles of the conscious cat].

Distribution, uptake and hydrolysys of cholesteryl oleate (-1-14C) in vivo in the cat's brain were investigated. Four hours following intraventricular injection of cholesteryl oleate (-1-14C), about one third of the injected radioactivity was taken up and about 50 % of that radioactive material was hydrolysed by brain tissue. Even more, the labelled material was found in the brain tissue surrounding the cerebral ventricles as well as in the cerebral cortex, but in varying amounts: the smallest amounts were found in the cerebral cortex, while the highest were in the hypothalamus. Radioactive material was also detected in the peripheral venous blood.

Animals↗

Substance P-like immunoreactivity in the trigeminal ganglion. A fluorescence, light and electron microscope study.

The trigeminal ganglion of rat and guinea pig was studied for the presence of immunoreactive substance-P using fluorescence, light and electronmicroscopy. In untreated animals substance P containing cells, with a diameter of 15 to 50 micron, were distributed throughout the ganglion and comprised 10-30% of all ganglion cells. Colchicine, injected intraventricularly to inhibit intra-axonal transport, had no effect on the number of substance P cells; but when the drug was injected directly into the posterior root of the ganglion, the proportion of these cells increased to as much as 50%. In the electron microscope, immunoreactive substance-P was confined to ganglion cells classified as B type according to the arrangement of subcellular organelles, and to unmyelinated nerve fibers. Subcellularly the immunoreactivity appeared in cytoplasmic vesicles, as well as dispersed in the nerve fibers and the perikarya of neurons. The great number of substance P immunoreactive ganglion cells suggests that they do not comprise a well defined subpopulation of the B-cells. However, the immunoreactivity was restricted to a distinct ultrastructural type of neurons with unmyelinated nerve fibers, suggesting that they also may share some distinct functions.

Animals↗

New method of obtaining cinchophen ulcer in dogs.

Cinchophen injected into the cerebral ventricle reacted with the hypothalamus somewhere in the vicinity ventricle, and produced a release of ACTH as indicated by a rise in peripheral plasma 11-OHCS concentration in the dog. The amount of cinchophen injected into the ventricle corresponds to about 1/600 of the dose required to produce the same effect by the intravenous route. Repeated intraventricular injections of cinchophen were effective in producing gastric erosions and ulcers at smaller doses than the intravenous one (1/100 and 1/400). Damage to the gastric wall might be due to a mechanism involving a localized portion of the hypothalamus and/or via a cerebral ventricle. Bilateral truncal vagotomy did not have any effect on chronic ulcer production. The cinchophen ulcerations were accompanied by an increase in plasma corticosteroids. The new experimental technique for producing peptic ulcer in dogs via a central neurohumoral mechanism is described.

11-Hydroxycorticosteroids↗

Effects of 6-hydroxydopamine and pimozide on temperature maintenance by the chicken.

Intraventricular implants of pimozide in adult white leghorn hens were used to block dopamine (DA) receptors, and 6-hydroxydopamine (6-OHDA) was injected intraventricularly to destroy the noradrenergic system locally. The hens were exposed to ambient temperatures of 5 and 35 degrees C, and their core temperature was measured. One hundred micrograms of 6-OHDA significantly reduced the norepinephrine (NE) but not the DA content of the hypothalamus and reduced the uptake of [3H]NE but not of [3H]DA by synaptosomes in vitro. Neither of the drug treatments nor their combination affected average core body temperature (Tb) at either 5 or 35 degrees C. Pimozide treatment caused a lower maximum Tb at 35 degrees C and a higher maximum Tb at 5 degrees C than the control treatment. No evidence was obtained that 6-OHDA treatment affected body temperature regulation. It is concluded that neither the DA nor the NE system is essential for normal temperature maintenance in the hen exposed to either 5 or 35 degrees C.

Animals↗