Methylated derivatives of l-tyrosine in reaction catalyzed by l-amino acid oxidase: isotope and inhibitory effects.

l-Amino acid oxidase (LAAO) is widely distributed in nature and shows important biological activity. It induces cell apoptosis and has antibacterial properties. This study was designed to investigate the effect of methyl substituent on its activity as methylated derivatives of l-tyrosine, labelled with short-lived B+ emitters, have been used in oncological diagnostics. To study isotope effects in the oxidative deamination of O-methyl-l-tyrosine, the deuterated isotopomer, i.e. O-methyl-[2-2H]-l-tyrosine, was synthesized by isotope exchange, catalyzed enzymatically by tryptophanase. Isotope effects were determined using the spectrophotometric non-competitive method. The values of isotope effects indicate that the α-C-H bond cleavage occurs in the rate determining step of the investigated reaction and α-hydrogen plays a role in the substrate binding process at the enzyme active site. The inhibitory effect on LAAO activity was studied with α-methyl-l-tyrosine and N-methyl-l-tyrosine. The mode of inhibition was determined based on Lineweavear-Burk plots intersections. α-Methyl-l-tyrosine has been found a mixed type inhibitor of the investigated enzyme, whereas N-methyl-l-tyrosine is a non-competitive inhibitor of LAAO.

Effects of the dopamine stabilizers (S)-(-)-OSU6162 and ACR16 on prolactin secretion in drug-naive and monoamine-depleted rats.

Dopaminergic stabilizers may be conceptualized as drugs with normalizing effects on dopamine-mediated behaviours and neurochemical events. (S)-(-)-OSU6162 (OSU6162) and ACR16 are two structurally related compounds ascribed such properties, principally because of their stabilizing effects on motor activity in rodents. Reports in the literature indicate possible partial D2 receptor agonist effects using various in vitro systems. This study aimed to measure D2 receptor antagonist and agonist effects of OSU6162 and ACR16 in vivo. To address this, we have studied the effects of both compounds on prolactin secretion in drug-naive and dopamine-depleted rats; dopamine depletion was induced by pretreatment with reserpine plus α-methyl-DL: -p-tyrosine. We find that OSU6162 and ACR16 both stimulate prolactin secretion in drug-naive rats with OSU6162 being considerably more potent and efficacious. Both compounds show a non-significant trend towards reversal of the increased secretion caused by dopamine depletion, whereas the D2 receptor antagonist haloperidol further increased prolactin secretion. Thus, this study suggests that OSU6162 and ACR16 act as D2 receptor antagonists under normal conditions in vivo, possibly with minor agonist effects in a state of dopamine depletion.

Requirement of a dopaminergic neuronal phenotype for toxicity of low concentrations of 1-methyl-4-phenylpyridinium to human cells.

LUHMES cells are conditionally-immortalized non-transformed human fetal cells that can be differentiated to acquire a dopaminergic neuron-like phenotype under appropriate growth conditions. After differentiation by GDNF and cyclic adenosine monophosphate, LUHMES were sensitive to 1-methyl-4-phenylpyridinium (MPP(+)) toxicity at < or =5 microM, but resistant to the parental compound 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). The high homogeneity and purity of the cultures allowed the detection of metabolic changes during the degeneration. Cellular ATP dropped in two phases after 24 and 48 h; cellular glutathione (GSH) decreased continuously, paralleled by an increase in lipid peroxidation. These events were accompanied by a time-dependent degeneration of neurites. Block of the dopamine transporter by GBR 12909 or mazindol completely abrogated MPP(+) toxicity. Inhibition of de novo dopamine synthesis by alpha-methyl-l-tyrosine or 3-iodo-l-tyrosine attenuated toxicity, but did not reduce the initial drop in ATP. Inhibition of mixed lineage kinases by CEP1347 completely prevented the MPP(+)-induced loss of viability and intracellular GSH, but failed to attenuate the initial drop of ATP. For the quantitative assessment of neurite degeneration, an automated imaging-based high content screening approach was applied and confirmed the findings made by pharmacological interventions in this study. Our data indicate that inhibition of mitochondrial ATP synthesis is not sufficient to trigger cell death in MPP(+)-treated LUHMES.

Inhibition of ATPases enzyme activities on brain disturbing normal oestrous cycle.

Wistar rats treated with alpha-methyl-DL-p-tyrosine methylester showed significant level of inhibition in the activity of Na+, K+ -ATPase, Mg2+ -ATPase and Ca2+ -ATPase enzymes in different regions of the brain. The enzyme activity was assayed in cerebral hemispheres, hypothalamus, thalamus, hippocampus, amygdala and septum at proestrous (12 h), estrous (25 h), metestrous (38 h) and diestrous periods (92 h) of the rat. The Na+, K+ -ATPase activity was significantly inhibited in most of the brain regions after treated with alpha-methyl-DL-p-tyrosine methylester (MPT) and this indicated that MPT affected the active transport system and nerve impulse transmission. Mg2+ -ATPase and Ca2+ -ATPase was also significantly (P < 0.001) reduced in different regions of the brain. The results revealed that MPT affected active transport system and nerve impulse transmission by inhibiting Na+, K+ -ATPase and Ca2+ -ATPase. It has induced energy crisis by inhibiting Mg2+ -ATPase and all these cumulative effects of MPT have adversely affected the female Wistar rats. These effects have been manifested in the form of aberrations in the behavior of MPT treated female rats, which have shown their inability to perform their normal sexual activity.

Striatal adenosine A(2A) receptor blockade increases extracellular dopamine release following l-DOPA administration in intact and dopamine-denervated rats.

The influence of the selective adenosine A(2A) receptor antagonist ZM 241385 on exogenous l-DOPA-derived dopamine (DA) release in intact and dopamine-denervated rats was studied using an in vivo microdialysis in freely moving animals. Local infusion of l-DOPA (2.5 microM) produced a marked increase in striatal extracellular DA level in intact and malonate-lesioned rats. Intrastriatal perfusion of ZM 241385 (50-100 microM) had no effect on basal extracellular DA level, but enhanced dose-dependently the l-DOPA-induced DA release in intact and malonate-lesioned animals. A non-selective adenosine A(2A) receptor antagonist DMPX (100 microM), similarly to ZM 241385, accelerated conversion of l-DOPA in intact and malonate-denervated rats. This effect was not produced by the adenosine A(1) receptor antagonist, CPX (10-50 microM). However, ZM 241385 did not affect the l-DOPA-induced DA release in rats pretreated with reserpine (5 mg/kg i.p.) and alpha-methyl-p-tyrosine (AMPT, 300 mg/kg i.p.). Obtained results indicate that blockade of striatal adenosine A(2A) receptors increases the l-DOPA-derived DA release possibly by indirect mechanism exerted on DA terminals, an effect dependent on striatal tyrosine hydroxylase activity. Selective antagonists of adenosine A(2A) receptors may exert a beneficial effect at early stages of Parkinson's disease by enhancing the therapeutic efficacy of l-DOPA applied exogenously.

Glutathione is involved in the granular storage of dopamine in rat PC 12 pheochromocytoma cells: implications for the pathogenesis of Parkinson's disease.

Parkinson's disease (PD) is characterized by degeneration of dopamine (DA)-containing nigro-striatal neurons. Loss of the antioxidant glutathione (GSH) has been implicated in the pathogenesis of PD. Previously, we showed that the oxidant hydrogen peroxide inhibits vesicular uptake of DA in nigro-striatal neurons. Hydrogen peroxide is scavenged by GSH and, therefore, we investigated a possible link between the process of vesicular storage of DA and GSH metabolism. For this purpose, we used rat pheochromocytoma-derived PC12 cells, a model system applied extensively for studying monoamine storage mechanisms. We show that depletion of endogenous DA stores with reserpine was accompanied in PC12 cells by a long-lasting, significant increase in GSH content the extent of which appeared to be inversely related to the rate of GSH synthesis. A similar increase in GSH content was observed after depletion of DA stores with the tyrosine hydroxylase inhibitor alpha-methyl-p-tyrosine. In the presence of alpha-methyl-p-tyrosine, refilling of the DA stores by exogenous DA reduced GSH content back to control level. Lowering of PC12 GSH content, via blockade of its synthesis with buthionine sulfoximine, however, led to a significantly decreased accumulation of exogenous [3H]DA without affecting uptake of the acetylcholine precursor [14C]choline. These data suggest that GSH is involved in the granular storage of DA in PC12 cells and that, considering the molecular characteristics of the granular transport system, it is likely that GSH is used to protect susceptible parts of this system against (possibly DA-induced) oxidative damage.

Inhibitory effects of putative dopamine D3 receptor agonists, 7-OH-DPAT and quinpirole, on prolactin secretion in rats.

The present experiments were performed to investigate effects of (+/-)-2-(dipropylamino)-7-hydroxy-1,2,3,4-tetrahydronaphthalene (7-OH-DPAT) or quinpirole (LY 171555), putative dopamine (DA) D3 receptor agonists, on serum prolactin levels in male rats. Basal prolactin levels were reduced dose-dependently by SC administration of 7-OH-DPAT or quinpirole at respective doses of 10-100 micrograms/kg and 25-250 micrograms/kg. Daily treatment with estradiol, 35 micrograms/kg/day for 3 days, increased serum prolactin levels to fourfold higher levels than those of nonprimed rats. Intraperitoneal injection of alpha-methyl-p-tyrosine (alpha-MT), 300 mg/kg, also increased serum prolactin levels. 7-OH-DPAT or quinpirole at a dose of 50 micrograms/kg caused a marked reduction in serum prolactin levels in both the estradiol- and alpha-MT-induced hyperprolactinemia. The 7-OH-DPAT- and quinpirole-induced decreases in serum prolactin levels were antagonized by the administration of the DA D2 receptor antagonist, spiperone, at 0.5 mg/kg. The results indicate that 7-OH-DPAT and quinpirole decrease prolactin levels in rats by stimulation of the D2 receptor.

Effects of administration of some monoamine-synthesis blockers and precursors on ovariectomy-induced rise in plasma gonadotropin II in the catfish Heteropneustes fossilis.

In the present study, effects of three daily i.p. injections of some monoamine (MA)-synthesis blockers and pre-cursors on plasma gonadotropin (GTH-II) levels were investigated in 3-week ovariectomized catfish, and the effects were correlated with changes in hypothalamic MA contents. Administration of alpha-methylparatyrosine (alpha-MPT; 250 micrograms/g BW, a tyrosine hydroxylase inhibitor) significantly decreased the ovariectomy-induced rise in GTH-II compared to that of the sham control group. Injection of L-dihydroxyphenylalanine (10 micrograms/g BW) in the alpha-MPT-treated fish elevated the GTH-II level significantly over that of the sham control group but not to the level of the ovariectomized fish. Administration of noradrenaline (NA; 5 micrograms/g BW) in combination with alpha-MPT counteracted the effect of the latter and maintained the plasma GTH-II level at that of the ovariectomized fish. Plasma GTH-II level was decreased significantly in the diethyldithiocarbamate (10 micrograms/g BW, a dopamine-beta-hydroxylase inhibitor)-injected fish compared to that of the sham and ovariectomized control groups. Administration of para-chlorophenylalanine (p-CPA; 100 micrograms/g BW, a tryptophan hydroxylase inhibitor) decreased the GTH-II level significantly compared to that of the ovariectomized group. Supplementation of 5-hydroxytryptophan (20 micrograms/g BW) with p-CPA nullified the latter's effect and maintained the GTH-II level at that of the ovariectomized fish. The administration of both p-CPA and alpha-MPT significantly reduced plasma GTH-II to the lowest mark compared to that of the sham, ovariectomized, and all other treatment groups. These results clearly show that the ovariectomy-induced rise in GTH-II was mediated through simultaneous activation of hypothalamic serotonergic and NA-ergic and suppression of dopaminergic mechanisms.

Mechanisms of altered LH secretion in neonatally oestrogenized male rats.

It is well known that the control of LH secretion depends on the steroid milieu during the postnatal period. In this study LH secretion was analysed in adult male rats injected neonatally with 500 micrograms oestradiol benzoate (1) after orchidectomy, (2) after selective elimination of androgens by destruction of Leydig cells with ethylene dimethane sulphonate (EDS), and (3) after removal in orchidectomized animals of Silastic capsules containing testosterone. In addition, (4) in vivo and in vitro LH secretion in response to LHRH agonist and antagonists, (5) the hypothalamic LHRH content, (6) the basal and stimulated in vitro LHRH release, and (7) the LH responses after administration of naloxone (2 mg/kg), alpha-methyl-p-tyrosine (alpha-MPT; 250 mg/kg), N-methyl-D-aspartic acid (NMDA, 15 mg/kg) or kainic acid (KA; 15 mg/kg) were also examined. Our data indicated that (1) the LH response after orchidectomy, after EDS administration and after removal of Silastic capsules containing testosterone was diminished in oestrogenized male rats, (2) the pituitaries from oestrogenized males retained responsiveness to LHRH, (3) hypothalamic LHRH content was reduced in oestrogenized males, but the hypothalamus from oestrogenized males released more LHRH than those of control groups both under basal conditions or after depolarization, (4) alpha-MPT decreased LH secretion only in oestrogenized males, and (5) NMDA and KA stimulated LH only in oestrogenized males. We conclude that in oestrogenized male rates the loss of sensitivity to the negative feedback action of testosterone on LH secretion was not due to decreased pituitary responsiveness to LHRH stimulation or to the inherent damage of LHRH neurones.(ABSTRACT TRUNCATED AT 250 WORDS)

Role of vesicular dopamine in the in vivo stimulation of striatal dopamine transmission by amphetamine: evidence from microdialysis and Fos immunohistochemistry.

The role of vesicular and newly synthesized dopamine in the action of amphetamine was investigated by studying the effect of reserpine and alpha-methyl-p-tyrosine pretreatment on amphetamine-induced changes in extracellular dopamine and acetylcholine, estimated by brain microdialysis, and on c-fos expression, estimated by quantitative immunohistochemistry of the Fos antigene, in the dorsal caudate-putamen of rats. Blockade of dopamine synthesis by alpha-methyl-p-tyrosine pretreatment (1 or 2 h) only partially prevented the increase in extracellular dopamine concentrations elicited by 0.5 and 2 mg/kg s.c. of amphetamine. Inactivation of vesicular amine uptake by reserpine pretreatment (3 h) reduced the increase in extracellular dopamine by 2 mg/kg but not by 0.5 mg/kg of amphetamine. Combined pretreatment with reserpine (3 h) and alpha-methyl-p-tyrosine (1 h) drastically reduced the increase in extracellular dopamine by both doses of amphetamine (0.5 and 2 mg/kg s.c.). alpha-Methyl-p-tyrosine pretreatment reduced c-fos expression stimulated by amphetamine (2 mg/kg) in the dorsomedial and dorsolateral caudate-putamen while reserpine pretreatment reduced it only in the dorsolateral caudate-putamen. Amphetamine (2 mg/kg s.c.) stimulated acetylcholine release but this effect was not modified by reserpine or alpha-methyl-p-tyrosine pretreatment. The results indicate that blockade of dopamine synthesis, by itself, is insufficient to prevent the stimulation of dopamine transmission by amphetamine and, conversely, that inactivation of vesicular dopamine significantly reduces amphetamine effects at pre- and postsynaptic levels. Therefore, vesicular dopamine appears to contribute to the stimulation of dopamine transmission elicited by amphetamine in the dorsal caudate-putamen.

Changes in hypothalamic catecholamines, dopamine-beta-hydroxylase, and phenylethanolamine-N-methyltransferase in the catfish Heteropneustes fossilis in relation to season, raised photoperiod and temperature, ovariectomy, and estradiol-17 beta replacement.

In Heteropneustes fossilis, contents and turnovers of hypothalamic catecholamines (CA) and activities of dopamine-beta-hydroxylase (DBH) and phenylethanolamine-N-methyltransferase (PNMT) showed significant seasonal variations with significantly high day values. The seasonal pattern of dopamine (DA) on one hand and that of noradrenaline (NA) and adrenaline (A) on the other hand showed an inverse relationship, the former decreasing and the latter increasing during the progress of gonadal recrudescence. The DBH and PNMT levels were low in the resting phase and increased to the peak in the prespawning (DBH) and spawning (PNMT) phases. Maintenance of the fish under long photoperiods (16L:8D) and high temperature (28 +/- 2 degrees) stimulated the NA and A, and DBH and PNMT activities, and suppressed the DA mechanism, the changes being maximal in the raised temperature groups. In the resting phase (December), ovariectomy (OVX) or estradiol-17 beta (E2) replacement in 4-week ovariectomized fish did not produce any significant effects on the CA and enzyme activities. On the contrary, in the prespawning phase (May), OVX produced differential and biphasic responses on CA and the enzymes. The contents and turnovers of both NA and A increased significantly at 2-5 weeks and decreased in the sixth week. However, the reverse was true for DA. The DBH and PNMT activities (assayed only 3, 4, and 6 weeks after OVX) were elevated significantly in the third and fourth weeks but decreased in the sixth week. Plasma levels of gonadotropin (GTH) increased significantly at all durations of OVX in a bimodal pattern while the E2 levels decreased consistently. Supplementation with a low dose (0.1 microgram/g BW) of E2 restored the NA and A and enzyme activities while the higher doses (0.5, 1.0, and 5.0 micrograms/g BW) depleted them. The reverse was true for DA. The low dose of E2 restored the GTH level while the higher ones inhibited it significantly. These results indicate that both environmental photoperiod and temperature and E2-negative feedback act on the CA to modulate GTH secretion.

In vivo actions of a gonadotropin-releasing hormone (GnRH) antagonist on gonadotropin-II and growth hormone secretion in goldfish, Carassius auratus.

In our previous in vitro studies, [Ac-delta 3-Pro1, 4FD-Phe2, D-Trp3,6]-mGnRH (analog E) suppressed both gonadotropin-II (GTH-II) and growth hormone (GH) release stimulated by sGnRH and cGnRH-II. In the present study analog E significantly inhibited the increases in plasma GTH-II levels stimulated by sGnRH in sexually mature female and sexually recrudescent goldfish. Treatment of goldfish with alpha-methyl-p-tyrosin methyl ester (alpha-MPT) inhibits dopamine synthesis and abolishes the inhibitory actions of dopamine on GTH-II release, resulting in a potentiation of the GTH-II response to sGnRH. Following alpha-MPT pretreatment, analog E significantly reduced basal plasma GTH-II levels, and suppressed both sGnRH and cGnRH-II actions on GTH-II release. Analog E also inhibited the increase in plasma GTH-II levels in sexually mature male goldfish exposed to the female sexual pheromone, 17 alpha, 20 beta-dihydroxy-4-pregnen-3-one (17 alpha 20 beta-P), demonstrating that the increase in plasma GTH-II levels is due to release of endogenous GnRH. Analog E significantly inhibited the increases in plasma GH levels stimulated by treatment with sGnRH. Implantation of estradiol pellets increases basal plasma GH levels and increases the GH responsiveness to sGnRH in sexually recrudescent goldfish; analog E also suppressed the increase in plasma GH levels stimulated by injection of sGnRH in estradiol-treated fish. Analog E suppressed basal GTH-II and GH levels in fish that were unhandled prior to injection; however, analog E was not effective in reducing basal plasma GTH-II or GH levels in experiments in which the fish were blood sampled or subjected to some experimental manipulation prior to injection of analog E. Analog E also suppressed basal levels of GTH-II in alpha-MPT-treated fish, suggesting that stress inhibition of GTH-II release may be mediated by the dopaminergic system. In summary, the results demonstrate that (i) analog E can suppress the actions of exogenous sGnRH and cGnRH-II on GTH-II and GH release in vivo, (ii) the GnRH system mediates, at least in part, the plasma GTH-II response in sexually mature male goldfish following exposure to the female sexual pheromone 17 alpha 20 beta-P, and (iii) endogenous GnRH peptides are important in the regulation of basal plasma levels of GTH-II as well as GH, particularly in low stress conditions.

Serotonergic involvement in the regulation of prolactin and vasoactive intestinal peptide mRNA expression in the rat anterior pituitary.

These studies examined the contribution of serotonin (5-HT) to the control of prolactin (PRL) and vasoactive intestinal peptide (VIP) messenger RNA expression in rat anterior pituitary. Daily injection of rats with the biosynthetic precursor to serotonin, 5-hydroxytryptophan (5-HTP; 25 mg/kg, q.i.d.), resulted on day 5 in a 50% increase in the expression of PRL mRNA in the pituitary while at the same time reducing the levels of both the 1.0 and 1.7 kb VIP mRNA transcripts. Co-treatment of rats with 5-HTP plus the catecholamine biosynthesis inhibitor, alpha-methyl-tyrosine (alpha-MT; 150 mg/kg, q.d. x 2 days), or the dopamine receptor antagonist haloperidol (1.25 mg/kg, b.i.d. x 5 days), resulted in increases in pituitary PRL message levels that were greater than those observed with either anti-dopaminergic agent alone. In contrast, 5-HTP was unable to reverse the inhibition of PRL mRNA expression caused by treatment with the dopamine receptor agonist bromocriptine (2.5 mg/kg, b.i.d. x 5 days). Neither alpha-MT, haloperidol nor bromocriptine had a significant effect on pituitary VIP mRNA expression. Administration of the direct-acting 5-HT receptor agonist quipazine (5 mg/kg, b.i.d.) for 14 consecutive days caused a significant increase in pituitary PRL mRNA levels on day 1 and reached a plateau of 90% above control levels on days 7 and 14. VIP mRNA levels rose significantly on day 1 of quipazine treatment but thereafter fell to a minimum of 22% (1.0 kb) and 52% (1.7 kb) of control by day 14.(ABSTRACT TRUNCATED AT 250 WORDS)

Newly discovered stereochemical requirements in the side-chain conformation of delta opioid agonists for recognizing opioid delta receptors.

Topographic design of peptide ligands using specialized topographically constrained amino acids can provide new insights into the stereochemical requirements for delta opioid receptors. A highly constrained tyrosine derivative, (2S,3S)-beta-methyl-2',6'-dimethyltyrosine [(2S,3S)-TMT], was prepared by asymmetric synthesis and incorporated in [D-Pen2,D-Pen5] enkephalin (delta 1) and Deltorphin I (delta 2). The results of binding assays and bioassays showed that the two analogues (3 and 4) acted very differently at delta opioid receptors. Further pharmacological evaluations suggested that they actually interact primarily with the delta 1 and delta 2 receptor subtypes, respectively. These results, and conformational studies using NMR and computer-assisted modeling, provided insights into the different stereochemical requirements for these two delta opioid ligands to recognize the delta opioid receptor and its subtypes.

Involvement of the catecholaminergic input to the paraventricular nucleus and of corticotropin-releasing hormone in the fasting-induced suppression of luteinizing hormone release in female rats.

The roles of the adrenergic projection to the paraventricular nucleus (PVN) and of central CRH in the suppression of pulsatile LH secretion during 48-h fasting were examined in ovariectomized estradiol (E2)-treated rats. The animals were ovariectomized and immediately implanted with Silastic tubing containing E2. One week after ovariectomy and E2 implantation, the animals were implanted stereotaxically with a guide cannula for microinjection into the PVN or intracerebroventricular (icv) injection. One week later, some of the animals were deprived of food for 48 h. The unfasted controls were provided with food ad libitum. At this point, blood samples were collected every 6 min for 3 h. Animals received an injection of 50 micrograms alpha-methyl-p-tyrosine (AMPT), a catecholamine synthesis inhibitor, into the PVN 3 h before the sampling started or an icv injection of 26 nmol alpha-helical CRF-(9-41), a CRH antagonist, after the first hour of blood sampling; control animals were given the vehicle at the equivalent time. The fasted animals injected with AMPT showed a significantly higher mean LH concentration and LH pulse frequency over the 3-h sampling period compared with the vehicle-injected controls. Treatment with AMPT had no significant effect on LH secretion in unfasted animals. The icv injection of alpha-helical CRF-(9-41) reinstated the suppressed LH release in fasted rats, but had no significant effect in unfasted animals. These results suggest that the adrenergic projection to the PVN and central CRH are involved in the suppression of pulsatile LH release during food deprivation. The possibility that fasting activates an ascending adrenergic projection that stimulates CRH release and thus suppresses pulsatile LH secretion is discussed.

Noradrenaline but not dopamine involved in NMDA receptor-mediated hyperalgesia induced by theophylline in awake rats.

Theophylline dose-dependently decreased a supraspinally integrated nociceptive threshold in awake rats. This hyperalgesia was antagonized by pretreatment with the N-methyl-D-aspartate (NMDA) receptor antagonist (+)-MK-801, suggesting involvement of NMDA receptors. Depletion of endogenous catecholamines with reserpine or alpha-methyl-DL-p-tyrosine and inhibition of noradrenaline synthesis with FLA 63 reduced the theophylline-induced hyperalgesia, whereas blockade of dopamine D2 receptors by pimozide, haloperidol (2 mg/kg) or (-)-sulpiride, of dopamine D1 receptors by SCH 23390, or of dopamine autoreceptors by a low dose of haloperidol (25 micrograms/kg), had no effect. By contrast, the alpha 1-adrenoceptor-blocking agent phenoxybenzamine abolished the hyperreactivity induced by theophylline, whereas the alpha 1-adrenoceptor antagonist prazosin and the beta-adrenoceptor antagonist (+/-)-propranolol were without effect. Furthermore, the alpha 2-adrenoceptor agonist clonidine (50 micrograms/kg) considerably decreased the hyperalgesia caused by theophylline. The adenosine A1/A2 receptor agonist N-ethyl-carboxamide adenosine (NECA) produced dose-dependent antinociception on the threshold for vocalization. Moreover, NECA (25 micrograms/kg) antagonized the hyperalgesia induced by different doses of theophylline, indicating that the effect is susceptible to purinergic modulation. It is suggested that theophylline-induced hyperreactivity to nociception is attributed to increased activity in NMDA and noradrenaline neurotransmission, possibly secondary to adenosine antagonism. Elevated intracellular levels of cyclic AMP might, however, also be involved in theophylline-produced hyperexcitability.

[Co-involvement of catecholaminergic and gabaergic transmission in the electro-cortical response to central galanin administration in the rabbit]. / Coinvolgimento della trasmissione catecolaminergica e gabaergica nella risposta elettrocorticale alla somministrazione centrale di galanina nel coniglio.

The involvement of catecholaminergic and gabaergic transmission in the electrocortical pattern induced by centrally administered galanin was investigated in the rabbit. The inhibition of catecholamine synthesis by both DDC and alphaMPT potentiated the EEG effect of the peptide. Baclofen and muscimol significantly enhanced the EEG response to GAL. However the interaction between the peptide and gabaergic transmission seemed mainly to involve the type A receptor system.

Thyrotrophin-releasing hormone-induced growth hormone secretion in domestic fowl: concomitant stimulation of dopamine turnover in the medial basal hypothalamus.

Thyrotrophin-releasing hormone (TRH) stimulates GH secretion in domestic fowl by actions at pituitary and central nervous system sites. The possibility that this central action might be mediated by hypothalamic catecholamines or indoleamines was therefore investigated. When TRH was administered into the lateral ventricles of anaesthetized fowl the concentration of 3,4-dihydroxyphenylacetic acid (DOPAC, a metabolite of dopamine (DA)) in the medial basal hypothalamus (MBH) was increased within 20 min. The concentrations of MBH noradrenaline (NA), DA, serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) were, however, unaffected by the intra-cerebroventricular (i.c.v.) administration of TRH, although the MBH concentrations of somatostatin and TRH were concomitantly reduced. A rapid increase in DA release into MBH extracellular fluid and its metabolism to DOPAC was also observed after i.c.v. or i.v. administration of TRH, in birds in which the MBH was perfused in vivo with Ringer's solution. Microdialysate concentrations of NA, 5-HT and 5-HIAA were not, however, affected by central or peripheral injections of TRH. Diminished GH responses to i.v. TRH challenge occurred in birds pretreated with reserpine (a catecholamine depletor), alpha-methyl-paratyrosine (a DA synthesis inhibitor) and pimozide (a DA receptor antagonist). These results therefore provide evidence for the involvement of a hypothalamic dopaminergic pathway in the induction of GH release following the central or peripheral administration of TRH. In contrast with its inhibitory actions at peripheral sites, DA would appear to have a central stimulatory role in regulating GH release in birds.