Search PubMed⌕ Search

Biomedical subjects

M Fillenz

Publications and source records attributed to M Fillenz.

At least 91 records · Page 5Linked to original sources

The development of linear sweep voltammetry with carbon paste electrodes in vivo.

A number of advances have been made in the application of linear sweep voltammetry with carbon paste electrodes to the monitoring of ascorbate and monoamine neuroregulator metabolites in the brain of the freely-moving rat. These new measures are: the reduction of the sweep rate of 5 mV/s; the elimination of semidifferentiation in chronic recordings; the measurement in situ and subtraction of the background current for each electrode; the reduction, to practically zero levels, of the ascorbate signal in alternate scans; and corrections for circadian changes. These innovations enable one to monitor changes in the monoamine metabolites, even when there are simultaneous, large changes in ascorbate.

Animals↗

The monitoring of ascorbate and monoamine transmitter metabolites in the striatum of unanaesthetised rats using microprocessor-based voltammetry.

Voltammetry in vivo has suffered from the limitation that electrodes which give high resolution of the various electroactive compounds present in brain tissue have a short life span, and, conversely, that stable electrodes give poor resolution. Carbon paste electrodes and linear sweep voltammetry with semidifferentiation can resolve changes in the concentration of ascorbate, deaminated and methylated metabolites of catecholamines, and the 5-hydroxyindoles in vivo and give stable recordings over many weeks. However, these changes in concentration are difficult to quantify. We have now developed the method further with the application of microprocessor technology. Microprocessor-based voltammetry and data analysis enable one to quantify changes in the concentration of electroactive species in complex mixtures where there are no separate oxidation peaks for the components; this has been achieved both in vitro and in vivo. Changes in dopamine release from the striatum have been monitored continuously over 48 h in unanaesthetised rats after the intraperitoneal administration of haloperidol. Carbon paste electrodes and microprocessor-controlled linear sweep voltammetry facilitates the quantitative, continuous monitoring of the extracellular concentration of ascorbate, the deaminated and methylated metabolites of catecholamines, and the 5-hydroxyindoles in the brain of unrestrained animals.

3,4-Dihydroxyphenylacetic Acid↗

The effects of footshock and handling on tyrosine hydroxylase activity in synaptosomes and solubilised preparations from rat brain.

The effects of mild noxious stimulation on central noradrenergic activity were investigated in rats, using low-intensity electrical footshock as the noxious stimulus. Synaptosomes were prepared from brain regions of control and experimental rats and the rate of tyrosine hydroxylation was assayed: this is a measure of transmitter synthesis in the isolated noradrenergic terminals. Single footshocks were found to elevate the synthesis rate in the hippocampus, the extent of the elevation depending on the baseline rate. This elevation was not significant in hypothalamus or cerebellum. Repeated handling over a period of 14 days was also found to elevate synthesis rate in hippocampus; but rats subjected to daily footshock for a week, in addition to handling, had synthesis rates similar to those of controls. Maximal tyrosine hydroxylase activity was measured by solubilizing the enzyme and assaying at saturating concentrations of cofactor and substrate: this is a measure of the amount of enzyme in the nerve terminals. No significant changes in maximal tyrosine hydroxylase activity were found in the brains of handled or shocked animals. These results suggest that the assay of tyrosine hydroxylase activity in a suspension of synaptosomes provides a sensitive indicator of changes in noradrenergic activity related to mild noxious stimulation. The biochemical responses are more pronounced in the hippocampus than in other noradrenergic terminal areas; this suggests that the synthesis rate of noradrenaline in terminals of central noradrenergic neurones could be regulated by local mechanisms at the terminal as well as by impulses transmitted from the cell body.

Animals↗

Circadian changes in homovanillic acid and ascorbate levels in the rat striatum using microprocessor-controlled voltammetry.

Circadian variations in the extracellular concentration of ascorbate and homovanillic acid in the striatum of unrestrained rats were measured simultaneously at 12 min intervals over periods of up to 48 h using carbon paste electrodes and microprocessor-controlled linear sweep voltammetry. Both compounds show a similar time course over a 24 h period, reaching a peak at 04.00 h and falling to a minimum at around 16.00 h. The changes in homovanillic acid probably reflect changes in dopamine release associated with circadian variations in the level of motor activity, while the origin and function of the ascorbate changes are unknown.

Animals↗

Lesions of the dorsal noradrenergic bundle and rewarded running: the role of pretraining.

Local injection of 6-hydroxydopamine was used to selectively destroy the dorsal ascending noradrenergic bundle (DB), producing 75% loss of hippocampal noradrenaline. Lesioned and control rats were trained to run in a straight alley for food reward with or without pretraining (handling and habituation to the apparatus). Lesioned rats ran more slowly than controls only if they had not been pretrained. This result may explain previous discrepancies in the literature; it is discussed in relation to existing hypotheses of DB function.

Animals↗

Acquisition and extinction of continuously and partially reinforced running in rats with lesions of the dorsal noradrenergic bundle.

Local injection of 6-hydroxydopamine was used to selectively destroy the dorsal ascending noradrenergic bundle (DB) in rats. Two lesion procedures were used, differing in the extent of depletion of forebrain noradrenaline they produced (greater than 90% or 77%). In Experiments 1-3 the rats were run in a straight alley for food reward on continuous (CR) or partial (PR) reinforcement schedules. The smaller lesion reduced and the larger lesion eliminated the partial reinforcement acquisition effect (i.e. the faster start and run speeds produced by PR during training) and the partial reinforcement extinction effect (PREE, i.e. the greater resistance to extinction produced by PR training); these changes were due to altered performance only in the PR condition. Abolition of the PREE by the larger DB lesion occurred with 50 acquisition trials, but with 100 trials the lesion had no effect. In Experiment 4 rats were run in a double runway with food reward on CR in the second goal box, and on CR, PR or without reinforcement in the first. The larger lesion again eliminated the PREE in the first runway, but did not block the frustration effect in the second runway (i.e. the faster speeds observed in the PR condition after non-reward than after reward in the first goal box). These results are consistent with the hypothesis that DB lesions alter behavioural responses to signals of non-reward, but not to non-reward itself. They cannot be predicted from two other hypotheses: that the DB mediates responses to reward or that it subserves selective attention. Since septal and hippocampal, but not amygdalar, lesions have been reported to produced similar behavioural changes, it is proposed that the critical DB projection for the effects observed in these experiments is to the septo-hippocampal system.

Animals↗

Linear sweep voltammetry with carbon paste electrodes in the rat striatum.

Voltammetry has been widely used in attempts to measure catecholamine release in vivo. The voltammogram recorded in the rat striatum using carbon paste electrodes and linear sweep voltammetry with semidifferentiation consists of a number of separate peaks; changes in the height of the first of these peaks have been attributed to changes in catecholamine release. We have found that ascorbate, either microinjected into the striatum or injected intraperitoneally, increases the height of the first peak without changing its potential. Microinjection of dopamine or 3,4-dihydroxyphenylacetic acid, or intraperitoneal injection of 3,4-dihydroxyphenylalanine, caused a shift in the potential of peak 1 of 25-50 mV in a positive direction. Amphetamine, administered intraperitoneally to freely moving animals, caused an increase in the height of the first peak but did not change its potential. Oxidation potentials in vitro and the effect of other drugs on the voltammogram obtained in vivo were also measured. Peak 1 is caused by the oxidation of both ascorbate and catechols whose oxidation potentials differ by only 50 mV in vivo; the contribution of catechols in control animals is negligible. Shifts in the potential of peak 1 caused by drugs are not due to changes in the oxidation potentials of the components but to a change in their relative contributions. Therefore changes in the height of peak 1 with no change in position do not represent changes in the extracellular concentration of catechols but are due to changes in ascorbate concentration. Changes in the concentration of catecholamine-related compounds can be detected at potentials some 50 mV greater than that of the first peak.

Animals↗

Parallel changes in ultrastructure and noradrenaline content of nerve terminals in rat vas deferens following transmitter release.

Transmural electrical stimulation and exposure to incubation media where some or all of the Na+ had been replaced with K+ were used to elicit transmitter release. Changes in noradrenaline content and ultrastructure of the nerve terminal varicosities in rat vas deferens were measured. Electrical stimulation in the presence of 4-aminopyridine had little effect, but high [K+] solutions caused a parallel reduction in noradrenaline content and the number of small dense-cored vesicles; large dense-cored vesicles showed no change, and small clear vesicles increased in number. In spite of a reduction in total vesicle number there was no evidence of expansion of the varicosity membrane. The parallel fall in noradrenaline content and in the number of small dense-cored vesicles suggests that the latter are the source of the released noradrenaline under the conditions of high [K+] stimulation we have used.

Animals↗

Control of noradrenaline release from hippocampal synaptosomes.

Potassium-evoked tritiated noradrenaline (NA) release from hippocampal synaptosomes was measured with a superfusion method. A single 2-min high-K+ pulse released 39% of the vesicular NA by a Ca2+-dependent mechanism: the Ca2+-independent release was negligible. After changing the vesicular NA store size by pretreating rats with either alpha-methyl-para-tyrosine, 500 mg/kg, or tranylcypromine, 10 mg/kg, a single K+ pulse released a constant percentage of the vesicular NA. With two K+ pulses, however, there was a reduction in the percentage of vesicular NA released in response to the second pulse.

Animals↗

Vesicular noradrenaline stores in peripheral nerves of the rat and their modification by tranylcypromine.

1 Vesicular noradrenaline stores were compared in the heart, salivary gland and vas deferens of the rat. 2 Noradrenaline storage vesicles in nerve terminals of different organs differed with respect to the amount of noradrenaline they contain in the endogenous store (content), the amount of exogenous noradrenaline they can take up from the circulation (uptake) and the amount of noradrenaline they contain when they are saturated (total storage capacity). 3 The data suggest that the vesicles in the salivary gland and vas deferens are almost completely filled with transmitter while, in the heart, the vesicular store is filled to only 55% of its total capacity. 4 The monoamine oxidase inhibitor, tranylcypromine, was found to increase not only the size of the endogenous store but also the size of the unfilled store.

Animals↗

Storage and release of noradrenaline in hypothalamic synaptosomes.

The noradrenaline storage capacity of vesicles in hypothalamic synaptosomes was measured by incubating them with [3H]noradrenaline under saturating conditions. The normal noradrenaline content is 52% of storage capacity. Incubation or superfusion with 50 mM-potassium causes calsium-dependent release from the vesicles. Such release reduces not only the vesicular content, but also the noradrenaline storage capacity. This suggests that after exocytosis vesicles cannot refill with noradrenaline.

Animals↗

Effect of runway training on rat brain tyrosine hydroxylase: differential effect of continuous and partial reinforcement schedules.

Previous experiments have implicated ascending noradrenergic systems in the development of the behavioural responses to different patterns of reward. In this report food deprived male Sprague--Dawley rats were trained to run a straight alley for good reward on a continuous reinforcement (CRF) or a partial reinforcement (PRF) schedule. Tyrosine hydroxylase measured in a partially solubilized preparation from hippocampus and hypothalamus at the end of acquisition was not different from controls, indicating that enzyme induction does not occur during either training schedules. However, hippocampal synaptosomal tyrosine hydroxylation rates from the CRF group was significantly higher than from either the PRF group or the handled controls. This indicates that at the end of the acquisition schedule the noradrenergic projection to hippocampus was more active in the CRF group than with the PRF group or the handled control.

Animals↗

Noradrenaline release in rats during prolonged cold-stress and repeated swim-stress.

1 Plasma noradrenaline concentration in rats was measured during prolonged cold-stress and repeated swim-stress. 2 Cold exposure for 6 h caused a rise in plasma noradrenaline which reached a peak at 4 h. 3 Administration of desmethylimipramine and normetanephrine to block neuronal and extra-neuronal uptake of noradrenaline raised plasma noradrenaline concentration without changing the pattern of the response to cold exposure. 4 Repeated cold exposure on subsequent days produced no change in the pattern of plasma noradrenaline concentration. 5 Five successive 1-min swims at 30-min intervals caused a rise in plasma noradrenaline concentration which was maximal after the third swim. 6 It is suggested that prolonged and repeated activation of sympathetic nerve terminals leads to a decline in noradrenaline release.

Animals↗