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Twice daily injections of 1.0 or 5.0 mg/kg d -amphetamine for 6 consecutive days differentially affected the response of dopaminergic neurons in the substantia nigra pars compacta to challenge injections of apomorphine on the following day. Thus, whereas treatment with the high amphetamine dose produced a dramatic shift to the right of the apomorphine dose-response curve, rats treated with 1.0 mg/kg d -amphetamine responded to apomorphine in the same way as saline-treated controls. These results support the view that long-term treatment with relatively high amphetamine doses is required to produce autoreceptor subsensitivity in the substantia nigra.

Circling behavior induced by dopamine (DA) agonists following microinjection of colchicine or 6-hydroxydopamine (6-OHDA) into the substantia nigra pars compacta (SNC) or electrolytic lesions of the SNC was investigated. Methamphetamine produced a contralateral circling behavior 3, 7 and 14 days following injection of colchicine into the SNC. Apomorphine produced an ipsilateral circling behavior followed by a contralateral rotation 3 and 7 days after the infusion of colchicine, whereas only an ipsilateral circling behavior was produced by apomorphine on day 14. 6-OHDA lesions of the SNC produced a contralateral circling behavior to apomorphine and an ipsilateral circling to methamphetamine, whereas both apomorphine and methamphetamine induced ipsilateral circling behaviors in rats with electrolytic lesions of the SNC. The possible mechanisms of action of colchicine are discussed in relation to the known effects of colchicine on axoplasmic transport.

Neuronal activity was recorded from the anteromedial neostriatum of rats pretreated twice daily for 6 consecutive days with saline or 5.0 mg/kg d-amphetamine. Glutamate was applied in both groups of animals to increase spontaneous firing rates. Iontophoretic application of increasing currents of DA (20–120 nA) produced a progressive inhibition of unit activity in control animals that was significantly reduced in rats pretreated with amphetamine. These results support the view that long-term amphetamine treatment reduces the sensitivity of postsynaptic DA receptors in the neostriatum.

The effects of chronic administration of methamphetamine on the responsiveness of neurons of the substantia nigra zona reticulata (SNR) to gamma-aminobutyric acid (GABA) or to a GABA receptor agonist were examined. Neuronal activity was recorded from the SNR of rats that had been pretreated twice daily, for 6 consecutive days, with saline or with 5 mg/kg methamphetamine. Intravenous administration of the GABA receptor agonist, muscimol, caused a dose-dependent decrease in the unit activity of the SNR neurons and the SNR neurons became less sensitive to the depressant effects of the drug after chronic treatment with methamphetamine. Iontophoretic application, with increasing currents, of GABA produced a progressive inhibition of unit activity in control animals, an effect that was significantly reduced in rats pretreated with methamphetamine. These results support the hypothesis that long-term administration of methamphetamine increases the activity of the striatonigral GABA system and thereby reduces the sensitivity of postsynaptic GABA receptors in the SNR.

The ability of methamphetamine to inhibit the firing rate of dopamine cells in the substantia nigra pars compacta (SNC) and the ventral tegmental area (VTA) was studied. Methamphetamine reduced the firing rates of the dopamine cells in a dose-dependent manner in the SNC and the VTA. The doses of methamphetamine required to produce a 50% inhibition of firing rate in the SNC and the VTA were 0.37 mg/kg and 0.28 mg/kg, respectively.

The electrical stimulation of the caudate nucleus could induce a caudate spindle in the rat as in case of the cat and monkey. The spindle was enhanced by atropine (3-10 mg/kg, i.v.) or morphine (3-10 mg/kg, i.v.) and the effect of these drugs were completely abolished by eserine (0.3 mg/kg, i.v.), but not by methysergide (1 mg/kg, i.v.), PCPA (400 mg/kg, i.p. for 3 days) or reserpine (3 mg/kg, i.p. for 2 days). An intraventricular injection of atropine (5-10 microgram/rat) or morphine (10 microgram/rat) exerted a facilitatory action on the spindle, while an intrathalamic and intracaudate injection of atropine (5-50 microgram/rat) showed a suppressive action. The spindle was also markedly enhanced by the microinjection of atropine (15-30 microgram/rat) or morphine (30 microgram/rat) into the reticular formation, and these actions were completely antagonized by eserine (0.3 mg/kg i.v.), but only partially by methysergide (4 microgram/rat, intraventricular administration.) These results suggest that the cholinergic system may play an important role in the regulation of the caudate spindle.

1. Acetylcholine (ACh)-induced relaxation of aortic strips with endothelium and production of cyclic GMP between streptozotocin-induced diabetic and age-matched control rats were compared. 2. The concentration-response curve for ACh-induced relaxation was shifted to the right in diabetic rats. IC50 values for ACh were 4.57 +/- 0.67 x 10(-8) M and 1.00 +/- 0.87 x 10(-7) M in aortic strips from age-matched control and diabetic rats, respectively (n = 6, P less than 0.05). 3. Relaxations produced by atrial natriuretic peptide (ANP) in diabetic aortae were similar to those in age-matched vessels. 4. Relaxations produced by sodium nitroprusside (SNP) in diabetic aortae were similar to those in age-matched vessels. 5. Basal levels of cyclic GMP and ACh-induced production of cyclic GMP were significantly decreased in diabetic rats. 6. These results suggest that functional changes in endothelium but not in guanylate cyclase activity in the aorta may occur in diabetes, and thus, spontaneous and ACh-induced formation of cyclic GMP may be decreased. This decrease in production of cyclic GMP may be responsible for the decreased response of the aorta to the relaxant effect of ACh.

An increase in the concentration of ascorbate in the extracellular fluid of the rat striatum following systemic amphetamine administration previously has been demonstrated with the use of in vivo electrochemistry, push-pull cannula and in vivo dialysis. In this report, the effect of infusions of amphetamine into the substantia nigra on extracellular ascorbate levels in the striatum is further investigated by in vivo electrochemistry. Electrolytic lesions in the crus cerebri of the striatonigral pathway abolish this effect. Furthermore, these lesions abolish the effect of systemic amphetamine in the striatum ipsilateral to the lesion but not on the contralateral side. This phenomenon is independent of neostriatal dopamine or gamma-aminobutyric acid since levels were shown to be unchanged relative to the intact side. These results lend further support to the hypothesis that ascorbate secretion in the striatum after amphetamine is mediated in the central nervous system, and indicate that a pathway that courses through the crus cerebri is necessary for this release.

Single-unit activity was recorded in the substantia nigra pars reticulata of rats in response to intravenous challenge injections of d -amphetamine. The animals were pretreated with saline or 5.0 mg/kg d -amphetamine twice daily for 6 consecutive days. Whereas the large majority of saline controls (6 to 8) showed no consistent response to amphetamine at doses up to 2.0 mg/kg, amphetamine pretreated rats (7 of 10) responded with a progressive increase in firing rate. Both groups of animals responded to a subsequent injection of 5.0 mg/kg clozapine with a depression of firing rate. The remaining control rats were inhibited by amphetamine and this aberrant response was enhanced with long-term treatment. In this unusual cells, clozapine accelerated firing rate. Taken together, these results indicate that unlike dopaminergic neurons in the compacta region of the nigra, reticulata neurons increase their responsiveness to amphetamine with repeated administration.

Voltammetric signals were recorded bilaterally with unmodified or electrochemically-modified carbon-fiber electrodes in the neostriatum of rats that sustained a unilateral depletion of dopamine by pretreatment with 6-hydroxydopamine. In both hemispheres, single daily injections of d -amphetamine (5.0 mg/kg or steadily increasing doses beginning with 6.0 mg/kg) for 6 consecutive days significantly reduced the rise in oxidation current produced by a challenge injection of 2.5 mg/kg d -amphetamine compared to the response obtained from saline-treated controls. Voltammetric scans indicated that the oxidized substance shared the electrochemical characteristics of ascorbic acid. It appears, therefore, that multiple amphetamine injections reduce the release of ascorbic acid in the neostriatum and that this effect does not depend on the integrity of dopaminergic neurons.

To investigate the influence of the striatonigral gamma-aminobutyric acid (GABA) system on the nigrostriatal dopamine (DA) system, the release of DA and/or 3,4-dihydroxyphenylacetic acid in the striatum ipsilateral to the injection side was examined by in vivo voltammetry following microinjection of GABA into the substantia nigra zona reticulata (SNR). The microinjection of GABA (100-300 micrograms/2 microliters) into the SNR produced a dose-dependent increase in the electrochemical signals recorded from the caudate nucleus ipsilateral to the injection side. Following chronic treatment with haloperidol, microinjection of GABA into the SNR produced only a slight (non-significant) increase in the electrochemical signals recorded from the caudate nucleus ipsilateral to the injection side. These results provide additional evidence to support the concept that DA cells in the substantia nigra zona compacta are regulated by the SNR non-DA neurons in an inhibitory manner. It appears, furthermore, that chronic treatment with haloperidol reduces the release of DA in the striatum ipsilateral to the injection side and that this effect may be due to a gradual development of depolarization block of DA cells by chronic administration of haloperidol.

Microelectrode and mechanical studies were performed with isolated guinea pig myocardium (right ventricular free walls and papillary muscles) to examine the effects of platelet-activating factor (PAF) and lysophosphatidylcholine (LPC). Low concentrations of PAF (10−8 to 10−6 M, a range equivalent to the blood concentrations that produce marked hypotension in vivo) had no effects on action potential configuration and contractile force. High concentrations (10−5 to 10−4 M) of PAF and LPC per se elicited slow response action potentials with concomitant contraction (restored contraction) in the myocardium depolarized with elevated K+ (25 mM); they also augmented slow responses and restored contractions produced by a low concentration of isoproterenol (10−8 M). Although these results suggested there was an increase in slow Ca current, the slow responses and restored contractions thus produced were greatly suppressed or abolished by the addition of a β-adrenoceptor blocking agent, sotalol (10−5 M), and by pretreatment with reserpine (5 mg/kg i.p., 24 h prior). In accordance with our previous conclusions, the present results suggest that direct cardiac action is not involved in the mechanisms of hypotension produced by PAF. It was also shown that high concentrations of PAF and LPC may act nonspecifically as amphiphilic compounds to induce transmitter release from sympathetic nerve endings, which may in turn augment the Ca current channels in the myocardial cell membrane.Key words: platelet-activating factor, cardiac action potential, slow response, Ca2+ channel, sympathetic nerve ending.

Phospholipase A2 (PLA2) from the venom of the snake Trimeresurus flavoviridis produced an increase in resting tension of isolated strips of rat stomach fundus. The contractions of the fundus strips induced by the PLA2 were significantly inhibited by treatment with 10(-6) M indomethacin and in Ca2+-free medium, while treatment of the fundus strips with nordihydroguaiaretic acid caused a marked potentiation of the PLA2-induced contraction. Atropine (10(-6) M), chlorpheniramine (10(-6) M) and methysergide (10(-6) M) had no effects on the contractions induced by PLA2, while tetrodotoxin (10(-6) M) significantly potentiated the contraction. From these results, it appears that exogenously applied PLA2 may cause contraction of the rat stomach fundus through the liberation of endogenous arachidonic acid which may then be transformed into prostaglandins.

Morphine induced-reduction in response to a repetitive electrical stimulation of the tail was measured in acute rats and rats chronically treated with haloperidol (1 mg/kg/d, for 7 d) following pretreatment with apomorphine. In acute experiments, a significant enhancement of the anti-struggling action of morphine was produced by haloperidol (1 mg/kg, i.p.). Low doses of apomorphine (30-480 micrograms/kg, i.p.) had no influence on the suppressing action of morphine on the struggling response induced by the tail stimulation. Following chronic treatment with haloperidol, the inhibitory action of morphine on the tail stimulation-induced struggling response was dose-dependently inhibited by very low doses of apomorphine (60-480 micrograms/kg, i.p.). A significant increase in 3,4-dihydroxyphenylacetic acid (DOPAC) levels was observed after administration of haloperidol or morphine in acute rats, whereas no change in DOPAC levels was found after administration of morphine or apomorphine in chronically haloperidol-treated rats. In these animals, basal striatal DOPAC levels were significantly decreased compared with those of vehicle-treated animals controls. The present results suggest that in rats treated chronically with haloperidol, the suppressive action of a low dose of apomorphine on morphine analgesia is due to an increased sensitivity of postsynaptic dopaminergic receptors to apomorphine.

1. To elucidate the mechanisms of the powerful and long-lasting hypotension produced by platelet activating factor (PAF), its effects on perfusion pressure in the perfused mesenteric arterial bed of the rat were examined. 2. Infusion of PAF (10(-11) to 3 x 10(-10) M; EC50 = 4.0 x 10(-11) M; 95%CL = 1.6 x 10(-11) - 9.4 x 10(-11) M) and acetylcholine (ACh) (10(-10) to 10(-6) M; EC50 = 3.0 +/- 0.1 x 10(-9) M) produced marked concentration-dependent vasodilatations which were significantly inhibited by treatment with detergents (0.1% Triton X-100 for 30 s or 0.3% CHAPS for 90 s). 3. Pretreatment with CV-6209, a PAF antagonist, inhibited PAF- but not ACh-induced vasodilation. 4. Treatment with indomethacin (10(-6) M) had no effect on PAF- or ACh-induced vasodilatation. 5. These results demonstrate that extremely low concentrations of PAF produce vasodilatation of resistance vessels through the release of endothelium-derived relaxing factor (EDRF). This may account for the strong hypotension produced by PAF in vivo.

SummaryIn obese patients with type 2 diabetes, insulin delivery to and insulin-dependent glucose uptake by skeletal muscle are delayed and impaired. The mechanisms underlying the delay and impairment are unclear. We demonstrate that impaired insulin signaling in endothelial cells, due to reduced Irs2 expression and insulin-induced eNOS phosphorylation, causes attenuation of insulin-induced capillary recruitment and insulin delivery, which in turn reduces glucose uptake by skeletal muscle. Moreover, restoration of insulin-induced eNOS phosphorylation in endothelial cells completely reverses the reduction in capillary recruitment and insulin delivery in tissue-specific knockout mice lacking Irs2 in endothelial cells and fed a high-fat diet. As a result, glucose uptake by skeletal muscle is restored in these mice. Taken together, our results show that insulin signaling in endothelial cells plays a pivotal role in the regulation of glucose uptake by skeletal muscle. Furthermore, improving endothelial insulin signaling may serve as a therapeutic strategy for ameliorating skeletal muscle insulin resistance.

A marked potentiation of the antinociceptive action of morphine was produced by haloperidol, but apomorphine had no effect on the antinociceptive action of morphine in acute experiments. Following chronic treatment with haloperidol, the antinociceptive action of morphine was significantly suppressed by apomorphine, and apomorphine shifted the dose-response curve of morphine to the right and increased the ED50 value of morphine by 2.3-fold. These results suggest that the suppressive action of apomorphine on the antinociceptive effect of morphine in chronic haloperidol-treated mice may be due to an increased sensitivity of post-synaptic dopaminergic receptors to apomorphine.

Rats were trained to lever-press for intracranial self-stimulation (ICSS) with electrodes implanted in the ventral tegmental area (VTA). The effect of inescapable footshock on response rates to ICSS was examined in the present study. Markedly decreased response rates to ICSS were observed 15 min to 24 h following inescapable footshock. Naloxone (10.0 mg/kg) itself was without effect on response rates to ICSS, but completely antagonized the decreased response rates by the stressor treatment. A relatively low dose of methamphetamine (0.5 mg/kg), which showed no effect on ICSS rates in naive rats, also antagonized the decreased response rates to ICSS. The present results suggest that inescapable footshock may release endorphin in the mesolimbic or mesocortical area; the released endorphin may act on dopaminergic nerve endings and interrupt dopaminergic transmission. The decreased activity of dopaminergic neurons may cause the decreased response rates to ICSS.

Effects of subchronic administration of haloperidol on the circling behavior induced by dopamine agonists were examined in rats treated with colchicine. Methamphetamine and apomorphine produced contralateral or ipsilateral rotation in colchicine-treated rats, and these circling behaviors were significantly increased by subchronic treatment with haloperidol. These results suggest that development of supersensitivity following colchicine and subchronic haloperidol may occur through different mechanisms.

Simultaneous recordings of single-unit activity and oxidation current were obtained bilaterally from the neostriatum of rats in response to a unilateral infusion of D-amphetamine (2.0 microliters of 10 micrograms/microliter) into the substantia nigra. Whereas neuronal activity increases ipsilaterally and decreases contralaterally, the electrochemical signals, which reflect extracellular ascorbate, increase bilaterally. In each case, changes in unit activity precede the change in ascorbate release. Systemic administration of haloperidol (0.2 mg/kg) blocks the increase in oxidation current bilaterally, but reverses the neuronal activity response only on the ipsilateral side. These results lend further support to the view that intranigral amphetamine facilitates neostriatal ascorbate release, but this effect is not correlated with changes in single-unit activity.

The effects of K+ activators on aortas from control and diabetic rats were examined. The concentration-response curves for the relaxant effects of cromakalim were shifted to the right in diabetic rats. The relaxation responses of diabetic aortas to nicorandil did not differ from those of the controls. Treatment with oxyhemoglobin significantly reduced the relaxation responses to nicorandil in aortas from diabetic rats. It appears that the activity of aortic potassium channels is reduced in diabetic rats.

Effect of microinjection of apomorphine into the substantia nigra zona reticulata (SNR) on the dopamine (DA) release in the ipsilateral striatum was examined in the present study. To measure the DA release, we determined DA and/or DOPAC levels in the striatum by using in vivo voltammetry. The microinjection of apomorphine (10 micrograms/2 microliters) into the SNR produced a rapid and prolonged increase in the DA and/or DOPAC peak in the ipsilateral striatum. These results suggest that release of DA from dendrites of DA cells normally plays a physiological role in the SNR neurons.

To investigate the influence of diabetes on the responsiveness of the cardiovascular system, we have examined the effects of various agents on the reactivity of the vascular smooth muscle of aortic strips obtained from age-matched control and diabetic rats. Norepinephrine (NE) contracted the aortic strips obtained from age-matched control and diabetic rats in a concentration-dependent manner, but maximal contraction of aortic strips in response to NE was enhanced in diabetic rats. The EC50 value for NE in the diabetic aortic strips was similar to that in the aorta from age-matched controls. Ca-induced contracture of the aortic strips which were depolarized with isotonic K+ (60 mM) was potentiated in aortic strips from diabetic rats, when compared with those from age-matched controls. Ca-induced contracture of aortic strips, preincubated with 10(-6) M NE and 10(-6) M nicardipine in Ca2+-free Krebs-Henseleit solution (KHS), was not significantly different in age-matched control and diabetic rats. Bay K 8644, an activator of calcium channels, produced an increase in the force of contraction of the slightly depolarized aorta from diabetic rats. Phasic contraction induced by phenylephrine (PE) in the presence of 10(-6) M nicardipine in Ca2+-free KHS was significantly enhanced in aortic strips obtained from diabetic rats. These results demonstrate that NE-induced contraction of the aortic strips obtained from diabetic rats was significantly enhanced, and that this increased contractile response of the aorta to NE may be due to an increased influx of extracellular calcium through the voltage-dependent Ca2+ channels, but not through the receptor-operated Ca2+ channels.(ABSTRACT TRUNCATED AT 250 WORDS)

Continuous rhythmic contractions were observed in longitudinal muscle strips of fundus obtained from the chronically vagotomized rat stomach. These vagotomy-induced rhythmic contractions (VRC) were not blocked either by tetrodotoxin (3 x 10(-6) M) or atropine (3 x 10(-7) M), indicating that the VRC was myogenic in origin. It was also found that the VRC had the following characteristics. 1) The VRC blocked either by 10(-5) M D600 or by Ca2+ deprivation from the medium. 2) On replacement of Na+ with sucrose or of K+ with Na+, the tone of fundus strips was concomitantly elevated with the cessation of the VRC. 3) 10(-5) M ouabain blocked the VRC. 4) The VRC was blocked by cooling. 5) The VRC disappeared under anoxic conditions. Based on these results possible mechanisms of the VRC are discussed in relation to an energy dependent activity of the cell membrane.

The characteristics of the non-adrenergic, noncholinergic inhibitory response of the rat stomach fundus to transmural nerve stimulation were compared with the relaxation induced by vasoactive intestinal polypeptide (VIP). Treatment with α-chymotrypsin (5 U/ml) or VIP antiserum (1:200) significantly reduced the relaxation induced by transmural nerve stimulation at 30 Hz, indicating that the possible transmitter in the non-adrenergic, non-cholinergic nerves is a peptide and may be VIP or a closely related peptide. VIP was able to relax, fully and dose-dependently, the stomach fundus that had previously been constricted by treatment with 10−6 M serotonin, and the IC50 value for VIP was 2.4 × 10−9 M. VIP elevated levels of cyclic AMP in a dose-dependent manner and the EC50 value was 2.8 × 10−9 M in the presence of 10−6 M atropine and 10−6 M guanethidine. The stomach fundus was relaxed by transmural nerve stimulation (30 Hz, 50 mA) and transmural nerve stimulation also caused production of cyclic AMP in the rat stomach in the presence of atropine and guanethidine. The basal level of cyclic AMP in the stomach was 8.7 ± 0.26 pmole/mg protein. When transmural nerve stimulation was applied for 5 min, the contraction of the stomach, induced by 10−6 M serotonin, was inhibited by 54% in the presence of atropine and guanethidine and the level of cyclic AMP was increased to 13.0 ± 0.73 pmol/mg protein. Apamin inhibited the transmural nerve stimulation-induced relaxation and shifted the dose-response curve for VIP to the right. These results suggest that one of the putative neurotransmitter from non-adrenergic, non-cholinergic nerves in the rat stomach is VIP and that VIP-induced relaxation may be mediated by the production of cyclic AMP and by the opening of apamin-sensitive K+-channels.

To determine the functional role of the striatonigral system in the circling behavior of rats and the mode of action of colchicine, we investigated the circling behavior induced by dopamine agonists after microinjection of colchicine into the unilateral striatum. Both apomorphine and methamphetamine produced ipsilateral circling behaviors in rats injected with colchicine, indicating that ipsilateral striatonigral pathways were damaged by the drug. Histological and biochemical examinations showed that intracaudate injection of colchicine damaged not only the dopaminergic neurons but also caused atrophy of the striatum with loss of neuronal perikarya. These results suggest that treatment with colchicine may be used as a model of senile atrophy or degenerative atrophy in these animals.

Circling behavior induced by unilateral microinjections of taurine and gamma-aminobutyric acid (GABA) into the substantia nigra zona reticulata (SNR) was studied in rats. Both taurine (10-100 micrograms/rat) and GABA (30-300 micrograms/rat) induced the contralateral circling behavior in a dose-dependent manner when they were microinjected into the SNR. Neither compound induced ipsilateral circling. An ipsilateral injection of 6-hydroxydopamine (6-OHDA) into the nucleus caudatus putamen (CPU) caused a reduction of the dopamine (DA) content in the injected side and decreased the circling behavior induced by taurine but not GABA. Electrolesion of the ipsilateral nucleus ventromedialis thalami (VM) decreased both taurine- and GABA-induced circling. In contrast, only GABA-induced circling was reduced by the electrolesion of the ipsilateral nucleus parafascicularis thalami (PF). Electrolesion of the ipsilateral nucleus accumbens (ACB) did not modify the circling behavior induced by GABA, but did decrease the circling induced by taurine. Electrolesion of the ipsilateral globus pallidus (GP) or nucleus entopeduncularis (EP) did not attenuate the circling behavior induced by either taurine or GABA. These results suggest that taurine-induced circling behavior may be mediated by the nigrostriatal-nigrothalamic system, and the ACB. In contrast, the nigrothalamic system may play an important role in GABA-induced circling behavior.

Neuronal activity was recorded from the ventral tegmental area (VTA) of immobilized, locally anesthetized rats on the day immediately following long-term treatment (twice daily for 6 consecutive days) with saline, 1.0 or 5.0 mg/kg d-amphetamine (d-AMPH). Each rat was challenged intravenously with d-AMPH (beginning with 0.0625 mg/kg) or with 0.005 mg/kg apomorphine. Treatment with d-AMPH significantly reduced the ability of this drug to inhibit VTA activity. In fact, nearly half of the neurons in the high-dose treatment group were excited by d-AMPH, whereas only 20% of control neurons showed this response. Moreover, apomorphine routinely accelerated firing rate in the VTA following treatment with 5.0 mg/kg d-AMPH but this response was never observed in control neurons, not even in those that were excited by d-AMPH. Thus, tolerance appears to develop to the ability of dopamine agonists to inhibit VTA activity and this effect may be mediated, at least in part, by a subsensitivity of inhibitory dopamine autoreceptors.