The spontaneous release of acetylcholinesterase in rat substantia nigra is altered by local changes in extracellular levels of dopamine.

Acetylcholinesterase release in the guinea-pig substantia nigra has been previously investigated 'on-line', using a sensitive chemiluminescent system. Since histological observations suggest that there is a difference in acetylcholinesterase distribution in the rat substantia nigra compared to that of the guinea-pig, the first aim of the present study was to use this chemiluminescent method to characterise acetylcholinesterase release in this brain region of the freely moving rat, and the second was explore the relationship between acetylcholinesterase release and dopamine systems in this region. Accordingly, acetylcholinesterase release in the rat substantia nigra was studied under basal conditions of spontaneous release and following the local administration of (a) elevated potassium ions (30, 45, 60'mM), (b) a stimulator of dopamine/acetylcholinesterase release-D-amphetamine (10(-7), 10(-6) and 10(-5) M), (c) an inhibitor of dopamine uptake-GBR12909 (10(-7), 10(-6) and 10(-5) M). Spontaneous release of acetylcholinesterase in this brain region of the rat appears to be comparable with that observed in the guinea-pig, despite the smaller number of acetylcholinesterase-containing neurones. Furthermore, not only elevated potassium ions, but D-amphetamine as well as GBR12909, all produced significant increases in the percentage spontaneous release of acetylcholinesterase. Thus, the release of acetylcholinesterase in this region may be triggered by levels of dopamine outside of the neurone.

Differential release of acetylcholinesterase in vivo, from the guinea pig substantia nigra compared to the caudate putamen following dopamine depletion.

In the substantia nigra acetylcholinesterase may have a novel role unrelated to acetylcholine but linked instead to dopamine. Using a sensitive chemiluminescent system, we have investigated the effects of dopamine depletion on the vivo release of acetylcholinesterase in both the substantia nigra and the caudate putamen. Dopamine levels in the caudate putamen were significantly depleted compared to the non-lesioned side, using either of two different toxins for dopaminergic nigrostriatal cells: 6-hydroxydopamine ( 1 or 3 weeks prior to study) or N-methyl-4-phenyl-1,2,5,6-tetrahydropyridine (1 week prior to study). Spontaneous release of acetylcholinesterase from the substantia nigra was significantly reduced following all three pretreatments; however, in the caudate putamen a significant reduction in the spontaneous release of acetylcholinesterase, compared to controls, was only seen in animals studied 1 week after the administration of 6-hydroxydopamine. In all control groups, application of potassium ions (60 mM) evoked a significant release of acetylcholinesterase in the substantia nigra (p < 0.05) and this effect persisted in the surviving neurones following a partial lesion by neurotoxin pre-treatment. The results from this study are discussed in the light of a regulatory mechanism for acetylcholinesterase release from the striatum, which may come into operation depending on the extent of destruction of dopaminergic nigrostriatal neurones.

Measurement of eicosanoid release in keratinocyte cultures to investigate skin irritation and tumour promoting activity.

Skin irritation, inflammation and hyperplasia appear to be intimately associated with the phenomenon of tumour promotion, but the mechanism of action remains elusive. Prostaglandins and leukotrienes play an important role in skin inflammation and prostaglandin E(2) (PGE(2)) modulates several events associated with phorbol ester-induced tumour promotion. This study investigated the release of eicosanoids (PGE(2) and leukotriene B(4)) and markers of cytotoxicity [neutral red (NR) uptake and intracellular acid phosphatase (AP) activity], after exposure of rat tongue epithelial (RTE) keratinocyte cultures to chemicals of different irritating and tumour promoting activity. The potent phorbol ester tumour promoter phorbol-12-myristate-13-acetate (PMA), and the less potent, structurally related diterpene ester, mezerein (MEZ) were compared with the known skin irritant sodium dodecyl sulfate (SDS). Cytotoxicity data reflected the in vivo skin irritation potential of the test chemicals and intracellular AP activity was increased after exposure to SDS (160 mug/ml), but did not appear to be increased by the more cytotoxic chemicals PMA and MEZ. Extracellular levels of PGE(2) were increased (200 to > 1000% of control levels) after an 18-hr exposure to PMA or MEZ over a concentration range of 0.01 to 20 mug/ml (NR(50) values 8.0 +/- 6.6 and 15.5 +/- 4.8 mug/ml, respectively). These data indicated that PGE(2) release occurred in the absence of cytotoxicity. In contrast, SDS only elicited PGE(2) release after exposure to 80 mug/ml (a cytotoxic dose level, NR(50) 82.5 +/- 9.9 mug/ml). The potency of a chemical to elicit PGE(2) release in keratinocytes in the absence of a cytotoxic response may reflect intracellular pathways intimately associated with the initiation of an inflammatory response and possibly with tumour promoting activity.

The release of acetylcholinesterase in vivo is regulated by dopaminergic systems in the guinea-pig substantia nigra.

Acetylcholinesterase (AChE) and dopamine are both stored and released from dendrites within the substantia nigra: however, it is as yet unknown whether the regulation of these two purported neuromodulators is in any way related. Using a sensitive chemiluminescent system to monitor AChE release 'on-line', the effects of inhibiting synthesis and storage of dopamine with alpha-methyl-p-tyrosine (AMPT: 250 mg/kg, i.p.) and reserpine (6 mg/kg, i.p.), respectively, have been studied. Both these agents significantly reduced nigral tissue dopamine levels by decreases of 83% and 63%, respectively; however, only AMPT had a significant effect in vivo on the spontaneous release of AChE compared to conscious control animals (66% decrease). Co-application of both AMPT and reserpine resulted in a significant decrease in the tissue dopamine content (95%) and in spontaneous release of AChE compared to conscious control guinea-pigs (72%); however, these effects were not significantly different from when AMPT was employed alone. Application of potassium ions (60 mM) or veratridine (100 microM) both evoked release of AChE in control animals: however, when expressed as a percentage of basal levels, this increase in release was not influenced by drug treatment or state of consciousness. These results suggest that de novo dopamine synthesis may at least in part, have an influential effect on release (and possibly storage) of AChE in the substantia nigra.

Effects of 6-hydroxydopamine on noradrenaline metabolism linked to neuronal uptake1 and extraneuronal uptake1 in dental pulp in vitro.

The study was undertaken to determine the relative roles of neuronal and extraneuronal uptake1 in the metabolism of 3H-noradrenaline in human dental pulp. Rabbit dental pulp was used as a reference since it was already known that normetanephrine (NMN) formation in this tissue utilised extraneuronal uptake1. Slices of pulp were preincubated in the absence and presence of 6-hydroxydopamine (1.6 mmol/l, for 10 or 20 min at pH 4.5) and subsequently incubated with 3H-noradrenaline (0.18 mumol/l for 30 min at pH 7.4). The principal metabolites formed were normetanephrine in rabbit pulp and deaminated catechols (dihydroxymandelic acid and dihydroxyphenylglycol) in human pulp. In both tissues 6-hydroxydopamine strongly inhibited formation of the deaminated catechols, but was without effect on normetanephrine formation. It is concluded that: i) in vitro 6-hydroxydopamine does not influence the metabolic process which is dependent on extraneuronal uptake1, namely normetanephrine formation in rabbit dental pulp, and ii) the deaminated catechols are largely neuronal in origin in human pulp. Attention is drawn to an unusual feature of the neuronal metabolism in human pulp, namely the appearance of dihydroxymandelic acid as the principal metabolite.

Petechial haemorrhage following a partial seizure with secondary generalization: atypical presentation and differential diagnosis.

A case is reported of a patient who had a respiratory arrest on a high dependency ward in a High Security Hospital with an unusual presentation. The patient had head and upper abdominal petechial haemorrhages with extensive conjunctival haemorrhaging. A considered antecedent for this potentially life-threatening presentation was strangulation. Analysis of all the available clinical information supports the hypothesis that he had a single tonic-clonic seizure with a focal-motor onset. This constitutes an unusual consequence of a partial seizure with secondary generalization.

Extraneuronal uptake of noradrenaline in rabbit dental pulp: evidence of identity with uptake1.

The extraneuronal removal and disposition of noradrenaline in rabbit dental pulp was examined in view of earlier evidence that the tissue possessed an extraneuronal uptake process resembling neuronal uptake1. Pulp, which had been depleted of sympathetic nerves by homolateral superior cervical ganglionectomy, was incubated in vitro with 3H-noradrenaline in low concentrations (0.025 or 0.18 mumol/l). When the metabolising enzymes (monoamine oxidase, catechol-O-methyl transferase) were active, 3H retention by the denervated pulp, as indicated by the 3H content after the tissue had been washed for 30 min following incubation with 3H-noradrenaline, was less than 3% of that of the innervated pulp. When the enzymes were inhibited, retention rose to approximately 30% of that of the innervated pulp. Analysis of the time course of the 3H efflux indicated that the 3H-noradrenaline in the denervated pulp had accumulated in a single compartment characterised by a t1/2 for efflux of several hours. Accumulation did not occur under Na(+)-free conditions, and was inhibited by desipramine (IC50 less than 0.03 mumol/l) and by substrates of neuronal uptake1. Mean IC50 values of the latter were very similar to those for inhibition of neuronal uptake1 and comprised (in mumol/l): (+)amphetamine (0.29), dopamine (0.31), tyramine (0.39), (-)noradrenaline (0.70), (-)adrenaline (1.50), 5-hydroxytryptamine (20) and bretylium (35). Uptake2 inhibitors were less active (O-methyl isoprenaline, IC50 = 60 mumol/l) than uptake1 inhibitors, or were without inhibitory effects at the concentrations tested (hydrocortisone, 210 mumol/l; 2-methoxy oestrone, 10 mumol/l).(ABSTRACT TRUNCATED AT 250 WORDS)

Serum lipids and eggs.

This study shows the variability of response of serum cholesterol to whole food dietary cholesterol eaten by free-living people. Divided into two groups in a crossover design, normolipemic, healthy volunteers in Group A showed statistically significant increases in mean serum cholesterol after eating three eggs daily for 10 weeks (p less than or equal to .01) and a significant decrease within 2 weeks of crossover to eating no eggs (p less than or equal to .03). Group B exhibited a significant decrease in mean serum cholesterol after eating no eggs for 12 weeks (p less than or equal to .02) but, after crossing over to eating 3 eggs daily for 10 weeks, showed no longitudinally significant increases. Only Group A showed statistically significant changes in mean high-density-lipoprotein cholesterol, i.e., an increase at the end of 12 weeks of eating three eggs daily (p less than or equal to .001). Mean serum triglycerides showed no significant change.