Biomedical subjects

K Pacak

Publications and source records attributed to K Pacak.

65 records · Page 4Linked to original sources

PubMed1994

Modification of cerebral cortical noradrenaline release by chronic inhibition of MAO-A.

Chronic treatment of rats with clorgyline (1 mg/kg i.p. daily for 21 days) caused a highly significant increase in the concentration of noradrenaline in microdialysate from the frontal cortex of the awake animal. Acute (one injection, 2 mg/kg) or subacute (1 mg/kg daily for 3 days) treatment did not lead to a significant increase in microdialysate noradrenaline. Concentrations of deaminated metabolites (DPHG, MHPG, DOPAC) in the microdialysate decreased with time of treatment, reaching a minimum after 21 days.

Animals↗

PubMed1994

Endogenous serotonin stimulates striatal dopamine release in conscious rats.

Serotonin (5-HT) and dopamine (DA) are established neurotransmitters in the brain. This study examined whether, in conscious, free-moving rats, increased concentrations of endogenous 5-HT in extracellular fluid of the corpus striatum affect local release of endogenous DA. Administration of the 5-HT reuptake blocker alaproclate via a microdialysis probe increased striatal dialysate levels of DA and its metabolites dihydroxyphenylacetic acid and homovanillic acid as well as levels of 5-HT and the 5-HT metabolite 5-hydroxyindoleacetic acid. Whereas DA reuptake blockade with GBR-12909 did not prevent these effects of alaproclate, serotonergic ablation by i.c.v. administration of 5,7-dihydroxytryptamine markedly decreased basal levels of 5-HT and 5-hydroxyindoleacetic acid and abolished the effects of alaproclate on dialysate levels of DA, 5-HT and their metabolites. The results are consistent with a stimulatory action of endogenous 5-HT on striatal DA release in conscious animals.

5,7-Dihydroxytryptamine↗

PubMed1993

Chronic inhibition of monoamine oxidase type A increases noradrenaline release in rat frontal cortex.

Chronic but not acute treatment of rats with MAO inhibitors, as with other antidepressant drugs, has been shown to down-regulate the number of cerebro-cortical beta-adrenoceptors. In order to establish whether this effect is associated with an increase in cortical noradrenaline release, rats were treated for 1, 3 or 21 days with clorgyline (2 mg/kg i.p. single injection; 1 mg/kg i.p. repeated injections), and the frontal cortex was then perfused by microdialysis in the awake animal. Control animals were injected with saline. The concentration of noradrenaline in the microdialysate increased only slightly after 1 or 3 days of clorgyline treatment but increased fourfold over control levels after 21 days treatment. Yohimbine (20 mumol/l) added to the perfusing solution caused a similar degree of enhancement in microdialysate noradrenaline concentration in all groups of rats. Tetrodotoxin (10 mumol/l) reduced noradrenaline concentration to low levels in all groups of animals, but noradrenaline was still detectable in the microdialysate in rats treated with clorgyline for 21 days. Concentrations of the deaminated metabolites dihydroxyphenylacetic acid, dihydroxyphenylglycol and methoxy-hydroxyphenylglycol were lowest after the 21 day clorgyline treatment. Determination of enzyme activity ex vivo showed that MAO-A was inhibited more than 95% by all clorgyline treatments with less than 10% inhibition of MAO-B. The results indicate that cerebrocortical noradrenaline release increases gradually during chronic MAO inhibition. This may be the result of more complete inhibition of the enzyme with time, not detectable by the ex vivo assay, but shown by the progressive reduction in metabolite levels.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1993

Role of CRH in glucopenia-induced adrenomedullary activation in rats.

Acute glucoprivation profoundly stimulated hypothalamic-pituitary-adrenocortical (HPA) and adrenomedullary outflows. Whether these responses reflect a single central mechanism regulated by corticotropin-releasing hormone (CRH) has been unclear. This study examined the role of endogenous CRH in HPA and adrenomedullary responses to hypoglycemia in Sprague-Dawley rats, by using anti-CRH immune serum or a CRH antagonist (alpha-helical h/r CRH9-41, and in Lewis rats, a strain characterized by deficient hypothalamic CRH responses during stress. In conscious Sprague-Dawley rats with indwelling arterial and venous cannulas, insulin (0.3 U/kg was injected iv, and responses of serum glucose concentrations and plasma levels of corticotropin (ACTH) and catechols (including epinephrine, EPI; norepinephrine, NE; dihydroxyphenylalanine, DOPA; dihydroxyphenylglycol, DHPG; and dihydroxyphenylacetic acid, DOPAC) were assessed, with or without pretreatment with anti-CRH immune serum (0.5 or 1.0 ml iv or 10 microl icv) or alpha-helical h/r CRH9-41 (130 nmol iv or 13 nmol icv). Responses to insulin (1.0 U/kg iv) were also measured in conscious juvenile Lewis and Fischer 344/N rats. Insulin-induced hypoglycemia markedly increased plasma levels of EPI and ACTH in all groups. Pretreatment iv with 1/0 ml of anti-CRH immune serum blocked the ACTH response to insulin but failed to attenuate the EPI response. alpha-helical h/r CRH9-41, whether given iv or icv, failed to alter ACTH or EPI responses to insulin, although the antagonist did block EPI responses to icv CRH. Hypoglycemia elicited similar increments in ACTH levels in Lewis rats and Fischer 344/N control rats; and although Lewis rats had lower baseline EPI and smaller responses of NE, DHPG, DOPA, and DOPAC levels, the groups did not differ in proportionate increments in EPI levels. The results indicate that the ACTH response to hypoglycemia depends on availability of CRH outside the blood-brain barrier--presumably in the pituitary gland. The findings with icv alpha-helical h/r CRH9-41 can be explained by failure of the antagonist to reach effective concentrations at central sites of action of endogenous CRH, or by mechanisms other than CRH release determining the adrenomedullary response to hypoglycemia. Lewis rats seem to have less adrenomedullary secretion at baseline and smaller responses of NE synthesis and release during hypoglycemia than do Fischer 344/N rats. Neurochemical evidence for differential adrenomedullary and sympathoneural responses during hypoglycemia in all three rat strains is inconsistent with Cannon's view of a functionally unitary sympathoadrenal system. Lewis rats have deficient CRH responses to some stressors but not to others, or else pituitary-adrenomedullary responses in this setting depend on mechanisms other than CRH release in the brain. Both explanations are inconsistent with the doctrine of non-specificity, the main tenet of Selye's stress theory.

Adrenal Medulla↗

PubMed1993

Glycine stimulates striatal dopamine release in conscious rats.

1. Glycine is an inhibitory neurotransmitter in the spinal cord and brainstem. The mechanism of this inhibition is via binding of glycine to specific receptors, increasing transmembrane Cl- conductance and hyperpolarizing neurones. Strychnine selectively antagonizes these effects. The role of glycinergic neurones in supraspinal regions is poorly understood. 2. Effects of glycine on release of catecholamines in the striatum were examined by microdialysis in freely-moving rats. Transcription of the genes encoding strychnine-sensitive glycine receptors was assessed in the striatum and substantia nigra, by use of reverse transcription followed by the polymerase chain reaction. 3. Glycine administered via the microdialysis probe dose-dependently increased concentrations of dopamine and its metabolites, dihydroxyphenylacetic acid and homovanillic acid, in the perfusate, indicating increased local release and metabolism of dopamine. Strychnine markedly attenuated these responses. Whereas striatal tissue did not contain mRNA for either the adult or neonatal form of strychnine-sensitive glycine receptor, nigral tissue contained a message for the adult form. 4. The results suggest that dopaminergic cells in the substantia nigra synthesize strychnine-sensitive glycine receptors and transport the receptors to terminals in the striatum. Occupation of the glycine receptors then exerts a net stimulatory effect on striatal dopamine release in vivo.

Animals↗

PubMed1993

Modified microdialysis probe for sampling extracellular fluid and administering drugs in vivo.

In vivo microdialysis provides an important new tool for investigating changes in extracellular fluid levels of endogenous compounds in vivo. Delivery of drugs via the microdialysis probe can be used to study local release and metabolism of neurotransmitters, but the dialysis membrane limits diffusion of substances between the perfusate and the extracellular fluid. Thus there may be considerable delay in responses, drug concentrations at the effector sites are less than those in the probe, and high-molecular-weight substances cannot traverse the membrane at all. This report describes a simple modification of commercially available microdialysis probes. A cannula is glued to the external surface of the probe. When glycine was administered via the cannula into the striatum of conscious rats, increments in microdialysate concentrations of dopamine were at least 10 times greater than when glycine was administered via the dialysis fluid in the probe. The threshold glycine dose for behavioral (turning) effects was also decreased by approximately 60-fold, and the time to the peak neurochemical and behavioral effects was markedly decreased. The modified probe did not destroy local catecholaminergic cells, as indicated by tyrosine hydroxylase immunofluorescence. Use of the modified microdialysis probe should facilitate pharmacological and neuroendocrine studies in behaving animals.

Animals↗

PubMed1993

Extraneuronal metabolism of endogenous and exogenous norepinephrine and epinephrine in rats.

Normetanephrine (NMN) and metanephrine (MN) are produced by the actions of catechol-O-methyltransferase on norepinephrine (NE) and epinephrine (E). Because catechol-O-methyltransferase is not present appreciably in neurons, plasma concentrations of NMN and MN provide a marker of extraneuronal catecholamine metabolism. In the present study, plasma NMN and MN were examined in rats before and during immobilization stress or i.v. infusion of NE and E to compare the extraneuronal metabolism of circulating and neuronally released catecholamines. At rest, plasma concentrations of NMN (0.44 +/- 0.03 pmol/ml) were 34% those of NE (1.40 +/- 0.11 pmol/ml) and MN concentrations (0.15 +/- 0.02 pmol/ml) were 33% those of E (0.48 +/- 0.11 pmol/ml). Immobilization and catecholamine infusion both increased plasma NMN and MN in parallel with increases in the precursor amines. Relative to increases in precursor amines, the MN response to infusion of catecholamines was 50% greater than the NMN response. Increases in plasma MN, relative to those in E, were similar during infusion of catecholamines and immobilization stress. In contrast, the NMN response to immobilization, relative to the NE response, was 3-fold larger than the NMN response to catecholamine infusion. Larger MN responses than NMN responses to infusion of catecholamines may reflect preferential extraneuronal uptake or O-methylation of E over NE or more efficient metabolism of NMN than MN. The larger NMN response to release of endogenous stores than to infusion of exogenous amine indicated that 30% of NMN is derived from circulating NE and 70% from NE in transit from sites of release to the circulation.

Adrenal Medulla↗

PubMed1992

Noradrenergic activation in the paraventricular nucleus during acute and chronic immobilization stress in rats: an in vivo microdialysis study.

In vivo microdialysis was used to study the effects of single (2 h) or repeated (2 h for 7 consecutive days) immobilization (IMMO) stress on extracellular fluid concentrations of norepinephrine (NE) and the deaminated metabolites of NE and dopamine, dihydroxyphenylglycol (DHPG) and dihydroxyphenylacetic acid (DOPAC) in the paraventricular nucleus of conscious rats. During IMMO, NE, DHPG, and DOPAC levels increased markedly, with similar peak values and time courses in the repeatedly stressed and previously unstressed groups. NE levels during a 2-h baseline period were lower in the repeatedly stressed group than in the unstressed group (99 +/- 9 pg/ml vs. 167 +/- 13 pg/ml, P less than 0.05), whereas DHPG (1,697 +/- 263 pg/ml vs. 1,424 +/- 194 pg/ml) and DOPAC (5,989 +/- 863 pg/ml vs. 4,428 +/- 1150 pg/ml) levels tended to be higher, so that the NE/DHPG ratio at baseline was significantly lower in the repeatedly stressed group (P less than 0.05). The results indicate that IMMO stress enhances NE release, reuptake, metabolism, and synthesis in the PVN. Repeated exposure to IMMO may decrease the microdialysate NE/DHPG ratio by inhibiting exocytotic release or enhancing neuronal reuptake of NE. In either case, the results suggest that repeated exposure to stress alters the release and disposition of NE in the PVN of conscious animals.

3,4-Dihydroxyphenylacetic Acid↗

PubMed1992

Hypercortisolemia inhibits yohimbine-induced release of norepinephrine in the posterolateral hypothalamus of conscious rats.

Chronic hypercortisolemia attenuates yohimbine (YOH)-induced increments in plasma levels of the sympathetic neurotransmitter norepinephrine (NE). The present study used in vivo microdialysis to study the effects of hypercortisolemia on YOH-induced release of NE in the brain. Cortisol (25 mg/kg.day) or saline was infused sc into rats for 7 days via an osmotic minipump. Microdialysate and plasma concentrations of NE and its metabolites dihydroxyphenylglycol and methoxyhydroxyphenylglycol were measured before and after YOH (1 mg/kg, iv) administration in conscious animals, with microdialysate and plasma collections beginning 20-24 h after probe implantation. Chronic cortisol treatment resulted in attenuated NE, dihydroxyphenylglycol, and methoxyhydroxyphenylglycol responses in both microdialysate and plasma. The results indicate that YOH increases central neural as well as peripheral release, reuptake, turnover, and metabolism of NE and that hypercortisolemia suppresses these responses.

3,4-Dihydroxyphenylacetic Acid↗

PubMed1991

Sympathoneural and skeletal muscle contributions to plasma dopa responses in pithed rats.

Dihydroxyphenylalanine (DOPA) in plasma has been thought to originate from sympathetic nerve endings and to reflect catecholamine biosynthesis, because changes in DOPA levels follow pharmacologically- or environmentally-induced manipulations that alter turnover of the sympathetic neurotransmitter, norepinephrine (NE). Skeletal muscle may be an additional, non-neural source of circulating DOPA. In the present study we examined sympathoneural and skeletal muscle contributions to DOPA in arterial plasma in pithed rats. Electrical stimulation of the spinal cord causes discharges of sympathetic post-ganglionic neurons, with attendant release of NE into the bloodstream, and discharges of spinal motoneurons, which causes diffuse contraction of skeletal muscle. Stimulation of the spinal cord rapidly elevated arterial plasma concentrations of NE, dihydroxyphenylglycol (DHPG), and DOPA. Pre-treatment with curare, a skeletal muscle relaxant, did not affect the NE and DHPG responses but attenuated the DOPA responses by about 50%. Administration of chlorisondamine, a ganglionic blocker, abolished NE and DHPG responses to cord stimulation, and DOPA responses were decreased by about 90%. Adrenal-demedullation did not affect the stimulation-induced DOPA responses. The results demonstrate that in pithed rats undergoing spinal cord stimulation, DOPA is released into the bloodstream. Since this response is markedly inhibited after ganglionic blockade and also attenuated after skeletal muscle paralysis, the results provide indirect evidence that DOPA formed in sympathetic neurons can be stored in a non-neuronal pool and released during skeletal muscle contraction.

Animals↗

PubMedDate not supplied

Disprocynium24, a novel inhibitor of the extraneuronal monoamine transporter, has potent effects on the inactivation of circulating noradrenaline and adrenaline in conscious rat.

The role of extraneuronal uptake in terminating the actions of catecholamines has been difficult to evaluate in vivo, largely because of lack of suitable inhibitors. The compound, 1,1'-diisopropyl-2,4'-cyanine iodide or disprocynium24 (D24), is a novel inhibitor of extraneuronal uptake with a high degree of potency in vitro. This study examined the actions of D24 on the inactivation and metabolism of circulating noradrenaline and adrenaline in conscious rats. Animals received i.v. infusions of 3H-labelled noradrenaline and adrenaline, and their extraneuronal O-methylated metabolites, normetanephrine and metanephrine. Plasma concentrations of endogeneous and 3H-labelled catecholamines and metanephrines were measured before and after D24. D24 caused large increases in plasma concentrations of noradrenaline and adrenaline, effects due to both decreases in their plasma clearances and increases in their rates of release into plasma. Plasma concentrations of normetanephrine and metanephrine also increased due to their decreased clearance from plasma. Increased release of normetanephrine into plasma did not contribute to increased plasma concentrations of normetanephrine. In fact, the contribution of extraneuronal O-methylation to noradrenaline clearance decreased substantially after D24. The data indicate that D24 is a potent inhibitor of the extraneuronal catecholamine transporter in vivo and that this process contributes importantly to the removal of circulating catecholamines and their O-methylated amine metabolites. Increased release of noradrenaline into plasma may reflect an increase in the proportion of transmitter that escapes from sites of release into the circulation. However, increased adrenaline release indicates that the drug also causes sympathoadrenal activation.

Adrenergic alpha-Agonists↗