Biomedical subjects

D H Versteeg

Publications and source records attributed to D H Versteeg.

At least 55 records · Page 3Linked to original sources

PubMed1989

Inhibition of noradrenaline release by antibodies to B-50 (GAP-43).

Protein kinase C (PKC) is believed to have a crucial role in synaptic transmitter release and long-term potentiation. An important substrate of PKC in the brain is the neuron-specific presynaptically localized protein B-50 (also termed GAP-43, F1, pp46 or P-57). B-50 has been implicated in the regulation of polyphosphoinositide metabolism and calmodulin binding, and in the mechanisms of neurite outgrowth, long-term potentiation and transmitter release. It is still unknown, however, whether B-50 (and/or its phosphorylation) is essential to any of these processes. Here we report the results of studies in which antibodies to B-50, which interfere with B-50 phosphorylation, were introduced into rat cortical synaptosomes that were permeabilized with streptolysin-O (SL-O). We found that the release of [3H]noradrenaline, induced by increasing the Ca2+ concentration in the buffer, is inhibited completely by the antibodies. These results provide the first demonstration of a causal relationship between the PKC substrate B-50 and the release of neurotransmitter.

Animals↗

PubMed1989

ACTH-(1-24) enhances the electrically stimulated release of [3H]dopamine from rat septal slices via a dopamine D2 receptor-independent mechanism.

ACTH-(1-24) enhanced the basal as well as the electrically stimulated release of [3H]dopamine from rat septal slices in vitro. In the absence of Ca2+ from the superfusion medium the effect of ACTH-(1-24) on the electrically stimulated release of [3H]dopamine was abolished. The stimulus-evoked release of [3H]dopamine from septal slices appeared to be modulated through dopamine receptors of the D2 subtype: the dopamine D2 receptor agonists 2-(N-propyl-N-2-thienylethylamino)-5-hydroxytetralin (N-0437) and quinpirole reduced, whereas the dopamine D2 receptor antagonist sulpiride enhanced the electrically stimulated release of [3H]dopamine. The magnitude of the effect of ACTH-(1-24) on [3H]dopamine release was the same in the presence or absence of N-0437, quinpirole and sulpiride. ACTH-(1-24) had no effect on either the basal or the electrically stimulated release of [3H]noradrenaline. Also when the electrically stimulated release of [3H]noradrenaline was reduced by the alpha 2-adrenoceptor agonist clonidine, the peptide was without effect. These results show that ACTH-(1-24) selectively enhances the release of [3H]dopamine from septal slices. The effect of the peptide is independent of the degree of activation of dopamine D2 receptors which modulate the stimulus-evoked release of [3H]dopamine. These results suggest that ACTH-(1-24) enhances the stimulus-evoked release of dopamine in the septum via a mechanism not associated with dopamine D2 autoreceptors.

Adrenocorticotropic Hormone↗

PubMed1989

Phosphorylation of B-50 (GAP43) is correlated with neurotransmitter release in rat hippocampal slices.

Recent studies have demonstrated that phorbol diesters enhance the release of various neurotransmitters. It is generally accepted that activation of protein kinase C (PKC) is the mechanism by which phorbol diesters act on neurotransmitter release. The action of PKC in neurotransmitter release is very likely mediated by phosphorylation of substrate proteins localized in the presynaptic nerve terminal. An important presynaptic substrate of PKC is B-50. To investigate whether B-50 mediates the actions of PKC in neurotransmitter release, we have studied B-50 phosphorylation in intact rat hippocampal slices under conditions that stimulate or inhibit PKC and neurotransmitter release. The slices were labelled with [32P]orthophosphate. After treatment, the slices were homogenized, B-50 was immunoprecipitated from the slice homogenate, and the incorporation of 32P into B-50 was determined. Chemical depolarization (30 mM K+) and the presence of phorbol diesters, conditions that stimulate neurotransmitter release, separately and in combination, also enhance B-50 phosphorylation. Polymyxin B, an inhibitor of PKC and neurotransmitter release, decreases concentration dependently the depolarization-induced stimulation of B-50 phosphorylation. The effects of depolarization are not detectable at low extracellular Ca2+ concentrations. It is concluded that in rat hippocampal slices B-50 may mediate the action of PKC in neurotransmitter release.

Animals↗

PubMed1988

Rapid postmortem increase in extracellular dopamine in the rat brain as assessed by brain microdialysis.

Extracellular dopamine (DA) and its metabolites 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in rat nucleus accumbens were determined before and shortly following death using microdialysis. A maximal 400-fold increase in the output of DA was observed within the first 5 min of death. DA output remained elevated over the following hour at a level of approximately 70-fold above pre-death values. In contrast to that of DA, DOPAC and HVA output gradually declined. Before death the extracellular DOPAC/DA ratio was about 250; after death this ratio dropped to 0.44 at 5 min. These observations may have important implications for experiments measuring the output of (endogenous) DA and its metabolites from brain tissue in vitro: autoregulation of, e.g., transmitter release and synthesis in vitro may be seriously disrupted by the observed depletion of transmitter storage granules.

3,4-Dihydroxyphenylacetic Acid↗

PubMed1987

The neuroleptic-like peptide desenkephalin-gamma-endorphin does not antagonize the dopamine receptor agonist-induced inhibition of the release of [3H]dopamine from rat nucleus accumbens slices in vitro.

In rats, the non-opioid beta-endorphin (beta E) fragment desenkephalin-gamma-endorphin (DE gamma E, beta E6-17) antagonizes the hypomotility induced by a small dose of dopamine (DA) receptor agonists. It has been suggested that DE gamma E might act in this respect by a direct or indirect blockade of presynaptically located DA receptors in the nucleus accumbens, thereby causing an increase of DA release. Therefore in the present study the effect of DE gamma E was examined on DA receptor agonist-induced inhibition of the electrically evoked release of previously accumulated [3H]DA from rat nucleus accumbens slices in vitro. The DA receptor agonists apomorphine, LY 171555 and n,n-di-n-propyl-7-hydroxy-2-aminotetralin (DP-7-AT) inhibited in a concentration-dependent manner the electrically evoked release of [3H]DA. The selective D2 receptor antagonist (-)-sulpiride blocked the effects of apomorphine, corroborating that the DA receptor involved is of a D2 type. DE gamma E was tested at several concentrations (10(-9)-10(-6) M) and under various experimental conditions. DE gamma E, by itself, did not affect either the electrically stimulated or the basal release of [3H]DA. The inhibiting effect of DA receptor agonists was slightly reduced by DE gamma E, but this effect was present in some experiments only. It is concluded that DE gamma E does not function as an antagonist for the DA receptor mediating DA release and that the interaction observed in behavioural experiments between DA agonists and DE gamma E does not occur at the level of this receptor.

Animals↗

PubMed1987

Microinjection of anti-vasopressin serum into limbic structures of the rat brain: effects on passive avoidance responding and on local catecholamine utilization.

Rats which had received bilateral microinjections of 1:50 diluted anti-vasopressin serum into the dorsal or ventral hippocampus, immediately after the learning trial of a one-trial passive avoidance test, showed a reduction in avoidance latency scores during subsequent retention tests 24 and 48 h later. Postlearning microinjection of anti-vasopressin serum into either the dorsolateral septum or the caudate nucleus was without effect on the retention of passive avoidance behavior. Microinjection of anti-vasopressin serum 1 h before the 24-h retention session into either the dorsal hippocampus, the ventral hippocampus or the dorsolateral septum attenuated avoidance responding during both the 24-h and 48-h retention sessions, whereas preretention microinjection of the serum into the caudate nucleus was not effective. Intracerebroventricular administration of the anti-vasopressin serum in amounts similar to those used in the microinjection experiments did not affect retention scores when given either immediately after the learning trial or before the first retention session. One week after the behavioral experiments, a repeated microinjection of anti-vasopressin serum decreased the local alpha-methyl-p-tyrosine methylester (alpha-MPT)-induced disappearance of noradrenaline in the ventral hippocampus and the dorsal hippocampus respectively. Microinjection of the antiserum in the dorsolateral septum enhanced noradrenaline disappearance in this brain region. No effect was found on alpha-MPT-induced dopamine disappearance in the caudate nucleus following local microinjection of anti-vasopressin serum.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1987

Basal and electrically stimulated release of [3H]noradrenaline and [3H]dopamine from rat amygdala slices in vitro: effects of 4 beta-phorbol 12,13-dibutyrate, 4 alpha-phorbol 12,13-didecanoate and polymyxin B.

The protein kinase C activator 4 beta-phorbol 12,13-dibutyrate (PDB) enhanced in a concentration-dependent manner the electrically stimulated release of [3H]noradrenaline ([3H]NA) and [3H]dopamine ([3H]DA) from rat amygdala slices in vitro. PDB enhanced the basal release of [3H]NA and [3H]DA as well. 4 alpha-Phorbol 12,13-didecanoate, which lacks the capacity to activate protein kinase C, was without effect on either basal or electrically stimulated release of [3H]NA and [3H]DA. Polymyxin B, which is a relatively selective protein kinase C inhibitor, decreased in a concentration-dependent manner the electrically stimulated release of both [3H]NA and [3H]DA from amygdala slices, whereas it enhanced the basal release of both neuromessengers. In the presence of 1.5 X 10(-7) M PDB, a concentration which when added to the superfusion medium alone doubled the electrically stimulated release of both [3H]NA and [3H]DA, polymyxin B again decreased in a concentration-dependent manner the release of both neuromessengers. At all polymyxin B concentrations used, the effect of the PKC inhibitor, expressed as percent inhibition, in the presence of PDB was approximately the same as that observed in the absence of PDB. This suggests that the antagonism between PDB and polymyxin B at the level of protein kinase C is not a competitive one. The effects of PDB and polymyxin B on basal release were additive. Taken together, these data suggest that in the amygdala presynaptically localized protein kinase C plays a role in signal transduction processes related to the exocytotic secretion of NA and DA from their nerve terminals.

Amygdala↗

PubMed1987

Phorbol 12,13-dibutyrate enhances electrically stimulated neuromessenger release from rat dorsal hippocampal slices in vitro.

The protein kinase C activator 4 beta-phorbol 12,13-dibutyrate (PDB) enhanced the electrically stimulated release of radiolabelled noradrenaline (NA), acetylcholine (ACh) and 5-hydroxytryptamine (5-HT) from dorsal hippocampal slices of the rat in vitro in a concentration-dependent manner. 4 alpha-Phorbol 12,13 didecanoate did not have an effect on the electrically stimulated release of any of the neuromessengers. Carbachol, which when present in the superfusion medium alone inhibited [14C]ACh release, significantly reduced the effect of PDB on the release of this neuromessenger. In the presence of either clonidine or [Leu5]enkephalin, which by themselves inhibited the electrically stimulated release of [3H]NA, the effect of PDB was significantly reduced. The enhancing effects of yohimbine and PDB on the electrically stimulated release of [3H]NA were additive. In all three cases, thus, the net effects of PDB were of a similar magnitude, whether the various compounds were present or not. Taken together, the present data suggest that the diacylglycerol/protein kinase C pathway is involved in the stimulus-evoked release of NA, ACh and 5-HT from dorsal hippocampal nerve terminals. Protein kinase C seems not to be involved in the modulation of the release of NA via presynaptic alpha 2-adrenoceptors and delta-opioid receptors and in that of ACh via presynaptic ACh receptors in that brain region.

Acetylcholine↗

PubMed1986

Facilitation of avoidance behavior by vasopressin fragments microinjected into limbic-midbrain structures.

Effects of arginine vasopressin (AVP1-9) and its behaviorally active fragments [Cyt6]AVP5-9 and [Cyt6]AVP5-8 were studied on the retention of one-trial learning passive avoidance behavior in rats. Peptides were microinjected into various limbic and midbrain structures (ventral or dorsal hippocampus or the dorsal raphe nucleus) and were administered either immediately after the learning trial (post-learning treatment) or shortly before the 24 h retention session (pre-retention treatment). Doses for intracerebral microinjections were selected after preliminary experiments with subcutaneous and intracerebroventricular peptide administration. AVP1-9 facilitated passive avoidance behavior when the peptide was microinjected into either brain structure, however, the ventral hippocampus appeared to be the most sensitive. In this limbic region, AVP1-9 facilitated passive avoidance behavior in an amount of 8 pg (bilaterally), both when given as post-learning or pre-retention treatment. [Cyt6]AVP5-9 and [Cyt6]AVP5-8 were more effective than the parent nonapeptide in terms that a lower amount of these peptide fragments facilitated passive avoidance behavior in all brain regions investigated. The ventral hippocampus appeared to be the most sensitive brain site for the behaviorally active vasopressin fragments as well. Following microinjections into the ventral hippocampus, [Cyt6]AVP5-8 was more effective in a post-learning than in a pre-retention treatment schedule. [Cyt6]AVP5-9 on the other hand was more effective when injected shortly before the retention trial. The data indicate that limbic-midbrain structures are sensitive to AVP1-9 and behaviorally active putative metabolites of this neuropeptide. The active fragments selectively influence different phases of information processing upon limbic microinjections.

Animals↗

PubMed1986

Brain dopamine depletion by lesions in the substantia nigra attenuates the development of hypertension in the spontaneously hypertensive rat.

The involvement of brain dopamine in the development of hypertension in the spontaneously hypertensive rat (SHR) was studied. Intracerebroventricular (i.c.v.) injections of 6-hydroxydopamine (6-OHDA) in young SHR caused depletion of dopamine in frontal cortex and striatum and induced an attenuation of the development of hypertension in SHR. Depletion of noradrenaline and to a lesser extent of serotonin was found as well. The ratio of DOPAC and of HVA to dopamine was increased after 6-OHDA. Pretreatment with the dopamine re-uptake inhibitor GBR-12909 inhibited the effects of 6-OHDA on both blood pressure and brain dopamine content. The effect of 6-OHDA on noradrenaline and serotonin levels were not influenced by pretreatment with GBR-12909. Electrolytic lesions in the substantia nigra delayed the rise in blood pressure in SHR. Lesions in the ventral tegmental area (VTA) were ineffective. After substantia nigra lesions depletion of dopamine was found especially in the nucleus caudatus posterior and the dorsomedial nucleus. After lesions in the VTA substantial dopamine depletion was found in the nucleus accumbens, frontal cortex, lateral septal nucleus and zona incerta. These data suggest that brain dopamine systems play a role in the development of hypertension in SHR and that especially the nigrostriatal system is important in this respect. Moreover, the present results may help to explain the attenuating effect of prehypertensive treatment with 6-OHDA on the development of hypertension.

Animals↗

PubMed1986

ACTH-(1-24) and alpha-MSH antagonize dopamine receptor-mediated inhibition of striatal dopamine and acetylcholine release.

The effects of ACTH-(1-24), alpha-MSH and ACTH-(4-10) were studied on the electrically evoked release of 3H-dopamine and 14C-acetylcholine from striatal slices in the absence and presence of the dopamine receptor agonist TL-99. None of the peptides affected transmitter release when TL-99 was not present. ACTH-(1-24) and alpha-MSH concentration-dependently antagonized the inhibition of striatal transmitter release induced by dopamine receptor stimulation due to the presence of TL-99. ACTH-(1-24), 10(-7)M, reduced the TL-99-induced inhibition of the release of both dopamine and acetylcholine by approximately 50%, and 5 X 10(-6) M ACTH-(1-24) restored the release fully to control values. alpha-MSH was less effective by a factor 20-30 in counteracting the release-inhibiting effect of TL-99. ACTH-(4-10) had no effect at any of the concentrations tested. These results show that ACTH/MSH-like neuropeptides may act by modulating dopamine receptor functions in rat striatum.

Acetylcholine↗

PubMed1986

Cardiovascular effects of central 6-OHDA treatment: a comparison of indirect and direct measurements.

The effect of intracerebro-ventricular treatment with 6-hydroxydopamine on blood pressure and heart rate was studied in spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto controls (WKY). When measured with the indirect tail-cuff method, the development of hypertension was found to be markedly inhibited in 6-OHDA treated SHR, while blood pressure was slightly lower in treated WKY. Heart rate was lower in both strains, although the greatest effect was found in SHR. In contrast, direct measurement via an arterial cannula indicated significantly lower blood pressure in 6-OHDA treated SHR only. Heart rate was by this method found to be not different between the SHR groups, but was increased in treated WKY. These results indicate that the mild stress of indirect blood pressure determinations has a marked influence on the results found.

Animals↗

PubMed1986

Substantia nigra lesions attenuate the development of hypertension and behavioural hyperreactivity in spontaneously hypertensive rats.

The possible relation between changes in behaviour and the development of hypertension was investigated. Depletion of striatal dopamine by lesions in the substantia nigra of Spontaneously Hypertensive Rats (SHR) was associated with an inhibition of the development of hypertension. In the open field a decrease in rearing score was found with no effect on other parameters. Rearing activity was significantly correlated with blood pressure as well as with striatal dopamine content. Blood pressure was weakly, although significantly, correlated with striatal dopamine content. Neither blood pressure nor striatal dopamine content was significantly correlated with ambulation activity. In normotensive Wistar-Kyoto rats a decrease was also found in rearing activity after nigra lesions, although this effect was less pronounced. Antihypertensive treatment of SHR with captopril or hydralazine did neither affect striatal dopamine levels nor open-field behaviour. Induction of renal hypertension or DOCA-salt hypertension in Wistar rats did not influence brain dopamine or behaviour. The results support the suggestion that brain dopamine systems may play a role in the development of hypertension in SHR as well as in the changes in behaviour observed in these rats. Changes in behaviour do not appear to be mediated by changes in blood pressure per se.

Animals↗

PubMed1986

Central 6-OHDA affects both open-field exploratory behaviour and the development of hypertension in SHR.

To investigate the possible relation between changes in behaviour and the development of hypertension in the spontaneously hypertensive rat (SHR), the effect of intracerebroventricular (ICV) administration of the catecholamine neurotoxin 6-hydroxydopamine (6-OHDA) on blood pressure and spontaneous behaviour was studied. In addition to an attenuation of the rise in blood pressure in SHR, 6-OHDA induced a marked decrease in rearing activity in the open field towards levels found in WKY. Other parameters were either not changed (stereotyped sniffing) or influenced in a comparable way in SHR and WKY (increase in locomotion). These results suggest that ICV 6-OHDA may simultaneously affect the development of hypertension and certain components of the changed behaviour of SHR. The exact relation between the two phenomena awaits further investigation.

Animals↗

PubMed1986

The dopamine metabolite 3-methoxytyramine is not a suitable indicator of dopamine release in the rat brain.

It has been postulated that changes in the concentration of 3-methoxytyramine (3-MT) in the brain might reflect changes in the release of 3,4-dihydroxyphenylethylamine (DA, dopamine) and, therefore, might be used as an index of dopaminergic activity in the brain. 3-MT is known to accumulate rapidly after death. Killing by microwave irradiation (MWR) is considered to be the method of choice to obtain "undisturbed" 3-MT concentrations. We measured striatal 3-MT concentrations even lower than those following MWR when the brains were excised and frozen in dry ice very rapidly (typical time between decapitation and freezing of the brain 22 s). There was a linear increase in striatal 3-MT concentration when the time between decapitation and freezing was varied between 13 and 300 s. Extrapolation to time zero indicated negligible amounts of 3-MT at the time of decapitation. In addition, it was observed that DA, 3,4-dihydroxyphenylacetic acid, and homovanillic acid decompose during the cooling phase after heating the brain by microwave. It is concluded that MWR induces artifactual changes in the postmortem levels of DA and metabolites. Consequently 3-MT cannot be considered to be a reliable indicator of DA release in the rat brain.

3,4-Dihydroxyphenylacetic Acid↗

PubMed1985

Interaction of vasopressin with the nigro-striatal dopamine system: site and mechanism of action.

The results of the present experiments show that local microinjections of Arg8-vasopressin into the nucl. caudatus cause an increase in the alpha-methyl-p-tyrosine methylester-HCl-induced disappearance of dopamine (DA) at the site of administration of the peptide. It is suggested that the caudate nucleus is the site of action of the peptide with respect to its effect on nigrostriatal DA neurons. This conclusion is corroborated by both the finding that microinjection of Arg8-vasopressin into the A9 region, which contains the cell bodies of the nigrostriatal system, was ineffective, and the results of push-pull experiments which showed an enhancement in apparent DA release in the nucl. caudatus when Arg8-vasopressin was co-perfused through the cannula system. Arg8-vasopressin appears to have a rather modest effect on nucl. caudatus DA synthesis, as was deduced from the results of experiments in which the in vitro conversion of tritiated tyrosine into tritiated DA was measured following in vivo Arg8-vasopressin administration as well as after in vitro incubation with the peptide. In conclusion, the interaction of vasopressin with the nigrostriatal DA system appears to be at the level of the DA terminals in the nucl. caudatus rather than at the level of the substantia nigra, and secondly, Arg8-vasopressin appears to affect DA release in the nucl. caudatus rather than DA synthesis.

Animals↗

PubMed1985

Effects of neonatal treatment with monosodium-glutamate in spontaneously hypertensive rats.

Following neonatal treatment with monosodium-glutamate (MSG), blood pressure in adult spontaneously hypertensive rats (SHR) was markedly lower than in control SHR, with only a small decrease in normotensive Wistar-Kyoto controls (WKY). In addition, in MSG-treated SHR an increase in pain-sensitivity was found and both strains showed increased water intake and reduced organ and body weights as compared to vehicle-treated rats.

Animals↗

PubMed1985

Digitoxin therapy partially restores cardiac catecholamine and brain serotonin metabolism in congestive heart failure.

The effect of therapeutic doses of digitalis in modifying neural activity has been the subject of considerable controversy. In earlier studies we reported an increase both in serotonergic activity in the posterior hypothalamus and pons-medulla and in cardiac sympathetic tone in the failing cardiomyopathic hamster. In this study we examine the effects of doses of digitoxin, known to be therapeutic for hamster heart failure, on monoamine neurotransmitter metabolism in the brain and heart during the cardiomyopathy. Both digitoxin and ASI-222, a polar amino-glycoside which does not cross the blood-brain barrier, given either acutely (6 mg/kg ip) or chronically (2 mg/kg/day ip for 10 days), normalized the failure-induced increase in serotonin turnover in the pons-medulla but had no effect on the changes in the posterior hypothalamus. Digitoxin therapy also reduced cardiac and adrenal sympathetic activity partially restoring cardiac catecholamine stores. In order to more clearly define the pathways involved we measured serotonin (microgram/g protein) in 18 brain nuclei after 10 days of digitoxin or vehicle treatment. Heart failure was associated with an increase in serotonin in five nuclei: the mammillary; bodies, ventromedial, periventricular and paraventricular nuclei of the hypothalamus, and the centralis superior nucleus of the raphe. Digitoxin therapy completely normalized the changes in the centralis superior and ventromedialis nuclei; neither congestive heart failure nor digitoxin affected serotonin levels in other nuclei. We conclude that there is an increase in activity in specific brain serotonergic nuclei in congestive heart failure. Digitalis reduces cardiac sympathetic tone and restores the changes in two of these nuclei: the ventromedial and the centralis superior.+2

Animals↗