Nitric oxide pathway in the nucleus raphe magnus modulates hypoxic ventilatory response but not anapyrexia in rats

Nucleus raphe magnus (NRM) is one of the cellular groups of the brainstem that is involved in the physiologic responses to hypoxia and contains nitric oxide (NO) synthase. In the present study, we assessed the role of NO pathway in the NRM on the hypoxic ventilatory response (HVR) and anapyrexia (a regulated decrease in body temperature). To this end, pulmonary ventilation (VE) and body temperature (Tb) of male Wistar rats were measured before and after microinjection of N-monomethyl-L-arginine (L-NMMA, a nonselective nitric oxide synthase inhibitor, 12.5 microg/0.1 microl) into the NRM, followed by hypoxia. Control rats received microinjection of saline. Under resting conditions, L-NMMA treatment did not affect pulmonary VE or Tb. Typical hypoxia-induced hyperventilation and anapyrexia were observed after saline treatment. L-NMMA into the NRM reduced the HVR but did not affect hypoxia-induced anapyrexia. In conclusion, the present study indicates that NO in the NRM is involved in HVR, exerts an inhibitory modulation on the NRM neurons but does not mediate hypoxia-induced anapyrexia.     

Nitric oxide in the rostral ventrolateral medulla modulates hyperpnea but not anapyrexia induced by hypoxia

Hypoxia causes hyperpnea and anapyrexia (a regulated decrease in body temperature, T(b)) but the mechanisms involved are not well understood. The nitric oxide (NO) pathway is involved in hypoxia-induced anapyrexia and hyperpnea, but the site(s) of action is not known. Nitric oxide synthase is present in the rostral ventrolateral medulla (RVLM), which is a nucleus in the medulla oblongata involved in control of breathing, and RVLM neurons have been suggested to have intrinsic hypoxic chemosensitivity. Therefore, we examined the effects of inhibition of the NO pathway in the RVLM on hypoxic hyperpnea and anapyrexia. Ventilation (VE) and body temperature (T(b)) were measured before and after bilateral microinjection of N-monomethyl-L-arginine (L-NMMA, 12.5 microg/0.1 microl, a nonselective nitric oxide synthase inhibitor) into the RVLM, followed by a 120-min period of hypoxic exposure. Control rats received microinjection of saline (vehicle). Under normoxia, L-NMMA treatment did not affect VE or T(b). Typical hypoxia-induced hyperpnea and anapyrexia were observed after saline treatment. L-NMMA treatment reduced the ventilatory response to hypoxia but did not affect hypoxia-induced anapyrexia. These data suggest that nitric oxide in the RVLM is involved in the ventilatory response to hypoxia, exercising an excitatory modulation of the RVLM neurons, but plays no role in hypoxia-induced anapyrexia.     

Role of central adenosine in the respiratory and thermoregulatory responses to hypoxia

No reports are available about the role of central adenosine in the respiratory and thermoregulatory responses to hypoxia in conscious rats. We therefore measured ventilation (VE) and body temperature (Tb) before and after intracerebroventricular injection of saline or aminophylline (adenosine antagonist), followed by a 30-min period of hypoxia exposure. Aminophylline did not change VE or Tb during normoxia; however, during hypoxia, it caused a significant increase in VE, and significantly attenuated hypoxic hypothermia. The present data indicate that central adenosine has an inhibitory effect on hypoxic hyperventilation and partially causes hypoxic hypothermia, suggesting that the ventilatory and metabolic interaction during hypoxia does not involve opposing mechanisms.     

Involvement of serotoninergic receptors in the anteroventral preoptic region on hypoxia-induced hypothermia

Hypoxia causes a regulated decrease in body temperature (Tb). There is circumstantial evidence that the neurotransmitter serotonin (5-HT) in the anteroventral preoptic region (AVPO) mediates this response. However, which 5-HT receptor(s) is (are) involved in this response has not been assessed. Thus, we investigated the participation of the 5-HT receptors (5-HT1, 5-HT2, and 5-HT7) in the AVPO in hypoxic hypothermia. To this end, Tb of conscious Wistar rats was monitored by biotelemetry before and after intra-AVPO microinjection of methysergide (a 5-HT1 and 5-HT2 receptor antagonist, 0.2 and 2 microg/100 nL), WAY-100635 (a 5-HT(1A) receptor antagonist, 0.3 and 3 microg/100 nL), and SB-269970 (a 5-HT7 receptor antagonist, 0.4 and 4 micro/100 nL), followed by 60 min of hypoxia exposure (7% O2). During the experiments, the mean chamber temperature was 24.6 +/- 0.7 degrees C (mean +/- SE) and the mean room temperature was 23.5 +/- 0.8 degrees C (mean +/- SE). Intra-AVPO microinjection of vehicle or 5-HT antagonists did not change Tb during normoxic conditions. Exposure of rats to 7% of inspired oxygen evoked typical hypoxia-induced hypothermia after vehicle microinjection, which was not affected by both doses of methysergide. However, WAY-100635 and SB-269970 treatment attenuated the drop in Tb in response to hypoxia. The effect was more pronounced with the 5-HT7 antagonist since both doses (0.4 and 4 microg/0.1 microL) were capable of attenuating the hypothermic response. As to the 5-HT(1A) antagonist, the attenuation of hypoxia-induced hypothermia was only observed at the higher dose. Therefore, the present results are consistent with the notion that 5-HT acts on both 5-HT(1A) and 5-HT7 receptors in the AVPO to induce hypothermia, during hypoxia.     

Hypothermia abolishes hypoxia-induced hyperpermeability in brain microvessel endothelial cells

The effect of mild (32 degrees C) and deep (22 degrees C) hypothermia on hypoxia-induced hyperpermeability was examined using an in vitro model of brain derived microvascular endothelial cells (BMEC). It was shown that hypoxia-induced hyperpermeability to inulin across the BMEC monolayer was completely abolished at 32 degrees C and 22 degrees C for up to 24 h of hypoxia. During normoxia, no influence of hypothermia on BMEC monolayer permeability was observed. The hypoxia-induced decrease of the cyclic AMP level after 6 h was abolished at 32 degrees C as well as at 22 degrees C of hypoxia. But after 24 h of hypoxia, hypothermia did no longer prevent the hypoxia-induced decrease of the cAMP level, which suggests that the effect of hypothermia on hypoxia-induced hyperpermeability is not caused by maintenance of the cAMP level. Because vascular endothelial growth factor (VEGF) has been shown to be the mediator of hypoxia-induced permeability changes of BMEC via the release of nitric oxide (NO), the effect of hypothermia on the VEGF expression was evaluated. During normoxia, hypothermia did not change the VEGF expression significantly but the hypoxia-induced increase in VEGF mRNA and protein expression was completely abolished at 32 degrees C and 22 degrees C respectively. Accordingly, the hypoxia-induced increase of the cGMP level was depressed by hypothermia, which demonstrates that also the amount of NO released during hypoxia is decreased at lower temperatures. Results suggest that deep as well as mild hypothermia decreased hypoxia-induced hyperpermeability by lowering the expression of the permeability-increasing protein VEGF and with it the release of NO.     

NO synthase inhibitors reduce opioid desensitization in rat locus coeruleus neurons in vitro.

The aim of this study was to examine by electrophysiological techniques whether nitric oxide (NO) is involved in the development of desensitization to the opioid agonist Met5-enkephalin (ME) in locus coeruleus neurons from rat brain slices. Bath perfusion with ME (0.05-1.6 microM) caused a concentration-dependent reduction in the firing rate of locus coeruleus cells, whereas perfusion with a high concentration of ME (10 microM) desensitized the inhibitory effect of subsequent ME (0.8 microM) applications. However, in slices perfused with the NO synthase inhibitors 7-NI (100 microM), L-NAME (100 microM) or L-NA (100 microM) the ME (10 microM)-induced opioid desensitization was strongly attenuated. The effect of L-NAME was prevented by administration of L-arginine (100 microM). These results suggest that nitric oxide may contribute to opioid desensitization in locus coeruleus neurons.     

Role of nitric oxide, adenosine,N-methyl-d-aspartate receptors, and neuronal activation in hypoxia-induced pial arteriolar dilation in rats

In this study, we tested the hypothesis that nitric oxide (NO) and adenosine (ADO) are the principal mediators of severe hypoxia-induced vasodilation. In addition, we examined whether activation ofN-methyl-d-aspartate (NMDA) receptors and/or perivascular nerves plays a role. A closed cranial window and intravital microscopy system was used to monitor diameter changes in pial arterioles (∼ 40 μm) in anesthetized rats. The relative contributions of ADO, NMDA, NO, and neuronal activation to hypoxic cerebrovasodilation were assessed using the blockers 8-sulfophenyltheophylline (8-SPT), MK-801, nitro-l-arginine methylester (LNAME), and tetrodotoxin (TTX). Two experimental series were studied. In the first, we tested the effects of NOS inhibition, via topical L-NAME (1 mM), on moderate (PaO₂ ≈ 46 mmHg)then severe (PaO₂ ≈ 34 mmHg) hypoxia-induced dilation. To confirm that L-NAME was affecting specifically NO-dependent responses, we also examined, in each experiment, the vasodilatory responses to topical applications of NOS-dependent (adenosine diphosphate (ADP); acetylcholine (ACh)(and -independent (sodium nitroprusside (SNP)) agents, in the presence of L-NAME or, in controls, the presence of D-NAME or no added analogue. In the second series, topical suffusions of ADP, ADO, and NMDA were sequentially applied, followed by 5 min exposure to severe hypoxia (PaO, ≈ 32 mmHg). Following return to normoxia, a suffusion of either 8-SPT (10 μM), MK-801 (10 μM), TTX (1 μM), or 8-SPT + MK-801 was initiated (or, in controls, application of a drug-free suffusate was maintained), and the above sequence repeated. In control, TTX, and 8-SPT + MK-801 experiments, baseline conditions were then restored and hypercapnia (PaCO₂ = 70–85 mmHg) was imposed. In the series 1 control groups, moderate and severe hypoxia elicited ≈ 20% and 35–40% increases in diameter, respectively. L-NAME attenuated ADP- and ACh-induced dilations, did not alter the arteriolar responses to SNP or moderate hypoxia, but prevented further dilation upon imposition of severe hypoxia. This suggested that 45–50% of the severe hypoxia response was NO-dependent. In series 2, 8-SPT blocked the adenosine response and reduced severe hypoxia-induced dilation by 46%. MK-801 predictably blocked NMDA-induced relaxation and reduced the hypoxic response by 42%. When combined, 8-SPT and MK-801 affected hypoxic vasodilation additively. After TTX, the ADP and ADO responses were normal, but NMDA and hypoxia responses were completely blocked. Hypercapnia-induced dilation was unaffected by TTX or 8-SPT + MK-801. The results imply that severe hypoxia-induced release of NO and ADO, and the accompanying pial arteriolar dilation, are wholly dependent on the capacity to generate action potentials in perivascular nerves. The similarity of the L-NAME and MK-801 effects on hypoxic cerebrovasodilation suggests that the NO-dependency, to a large degree, derives from NMDA receptor activation.     

Role of nitric oxide in 2-deoxy-D-glucose-induced hypothermia in rats.

The present study was designed to test the hypothesis that nitric oxide (NO) plays a role in 2-deoxy-D-glucose (2-DG)-induced hypothermia. The body temperature of awake, unrestrained rats was measured before and after the administration of 2-DG, or N(G)-nitro-L-arginine methyl ester (L-NAME; a non-selective NOS inhibitor) or both treatments together. We observed a significant reduction in body temperature after 2-DG injection. L-NAME alone caused no significant change in body temperature. When the two treatments were combined, a reduction in the magnitude of 2-DG-induced hypothermia was observed. The neuronal NOS inhibitor 7-nitroindazole also inhibited 2-DG-induced hypothermia. The data indicate that NO, probably produced by neuronal NOS, plays a role in 2-DG-induced hypothermia.     

Role of nitric oxide, adenosine,N-methyl-d-aspartate receptors, and neuronal activation in hypoxia-induced pial arteriolar dilation in rats

In this study, we tested the hypothesis that nitric oxide (NO) and adenosine (ADO) are the principal mediators of severe hypoxia-induced vasodilation. In addition, we examined whether activation ofN-methyl-d-aspartate (NMDA) receptors and/or perivascular nerves plays a role. A closed cranial window and intravital microscopy system was used to monitor diameter changes in pial arterioles ( 40 μm) in anesthetized rats. The relative contributions of ADO, NMDA, NO, and neuronal activation to hypoxic cerebrovasodilation were assessed using the blockers 8-sulfophenyltheophylline (8-SPT), MK-801, nitro-l-arginine methylester (LNAME), and tetrodotoxin (TTX). Two experimental series were studied. In the first, we tested the effects of NOS inhibition, via topical L-NAME (1 mM), on moderate (PaO₂ ≈ 46 mmHg)then severe (PaO₂ ≈ 34 mmHg) hypoxia-induced dilation. To confirm that L-NAME was affecting specifically NO-dependent responses, we also examined, in each experiment, the vasodilatory responses to topical applications of NOS-dependent (adenosine diphosphate (ADP); acetylcholine (ACh)(and -independent (sodium nitroprusside (SNP)) agents, in the presence of L-NAME or, in controls, the presence of D-NAME or no added analogue. In the second series, topical suffusions of ADP, ADO, and NMDA were sequentially applied, followed by 5 min exposure to severe hypoxia (PaO, ≈ 32 mmHg). Following return to normoxia, a suffusion of either 8-SPT (10 μM), MK-801 (10 μM), TTX (1 μM), or 8-SPT + MK-801 was initiated (or, in controls, application of a drug-free suffusate was maintained), and the above sequence repeated. In control, TTX, and 8-SPT + MK-801 experiments, baseline conditions were then restored and hypercapnia (PaCO₂ = 70–85 mmHg) was imposed. In the series 1 control groups, moderate and severe hypoxia elicited ≈ 20% and 35–40% increases in diameter, respectively. L-NAME attenuated ADP- and ACh-induced dilations, did not alter the arteriolar responses to SNP or moderate hypoxia, but prevented further dilation upon imposition of severe hypoxia. This suggested that 45–50% of the severe hypoxia response was NO-dependent. In series 2, 8-SPT blocked the adenosine response and reduced severe hypoxia-induced dilation by 46%. MK-801 predictably blocked NMDA-induced relaxation and reduced the hypoxic response by 42%. When combined, 8-SPT and MK-801 affected hypoxic vasodilation additively. After TTX, the ADP and ADO responses were normal, but NMDA and hypoxia responses were completely blocked. Hypercapnia-induced dilation was unaffected by TTX or 8-SPT + MK-801. The results imply that severe hypoxia-induced release of NO and ADO, and the accompanying pial arteriolar dilation, are wholly dependent on the capacity to generate action potentials in perivascular nerves. The similarity of the L-NAME and MK-801 effects on hypoxic cerebrovasodilation suggests that the NO-dependency, to a large degree, derives from NMDA receptor activation.     

Distribution of c-fos mRNA in the brain following intracerebroventricular injection of nitric oxide (NO)-releasing compounds: possible role of NO in central cardiovascular regulation

We injected nitric oxide (NO)-releasing compounds and NO synthase (NOS) inhibitors into the brains of conscious, freely moving rats and measured the effects on mean arterial blood pressure (MAP) and heart rate, as well as on the expression of c-fos mRNA, neuronal NOS (nNOS) mRNA and NADPH-diaphorase, an indicator of NOS activity. When administered i.c.v., the NO donor, NOC-18, caused a significant fall in MAP and heart rate, whereas the NOS inhibitor, NG-nitro-L-arginine methyl ester (L-NAME), induced a significant rise in MAP. The same dose of NOC-18 or L-NAME when administered i.v. did not affect MAP and heart rate. Centrally administered NOC-18 induced c-fos mRNA expression in several regions of the brain involved in the baroreceptor response, including the nucleus of the solitary tract, the area postrema and the rostral ventrolateral medulla, as well as areas involved in the integration of autonomic, neuroendocrine and behavioural responses, including the medial preoptic area, the organum vasculosum lamina terminalis, the bed nucleus of stria terminalis, the paraventricular nucleus (PVN), the supraoptic nucleus (SON), the central nucleus of amygdala (CeA) and the locus coeruleus. Most of the areas that expressed c-fos also contained nNOS mRNA and/or NADPH-d-positive neurones and fibres. i.c.v. injection of L-NAME induced c-fos mRNA expression in PVN, SON, locus coeruleus and NTS, suggesting a tonic inhibition of neuronal activity by NO or stimulation of neuronal activity by endogenous NO. i.v. injection of NOC-18 or L-NAME did not induce any significant c-fos mRNA expression in rat brain. These results demonstrate that NO acts directly in the brain to reduce the systemic blood pressure, and that the endogenous NO pathway may play a role in cardiovascular and autonomic regulation by modulating neuronal activities in discrete regions of the brain.     

Effect of central and peripheral administration of a nitric oxide synthase inhibitor on morphine hyperthermia in rats

The effect of central and peripheral administration of a nitric oxide synthase inhibitor, N-nitro-L-arginine methyl ester (L-NAME), on morphine hyperthermia was studied in male Sprague-Dawley rats. The first series of experiments examined the effect of subcutaneous (s.c.) administration of L-NAME on the hyperthermia induced by morphine given s.c. in doses of 4 and 15 mg/kg. L-NAME, at a s.c. dose of 50 mg/kg, per se, had no influence on body temperature (T(b)). Coadministration of L-NAME (50 mg/kg, s.c.) with the higher dose of morphine (15 mg/kg, s.c.) caused a significant suppression of morphine hyperthermia during the first 30 min and then produced hypothermia. In contrast, s.c. injection of L-NAME (50 mg/kg, s.c.) failed to alter the hyperthermic response induced by the lower dose of morphine (4 mg/kg). In the second series of experiments, we investigated the effect of intracerebroventricular (i.c.v.) administration of L-NAME on the hyperthermia induced by morphine given s.c. L-NAME, itself, given i.c.v. at a dose of 1 mg did not evoke any change in T(b). Intracerebroventricular administration of L-NAME (1 mg) blocked the hyperthermia induced by 15 mg/kg morphine during the first 30 min and induced a slight hypothermia but did not alter the hyperthermia induced by 4 mg/kg morphine. The results indicate that either central or peripheral NO synthesis is required for the production of hyperthermia induced by 15 mg/kg of morphine. However, NO synthesis does not seem to be involved in the hyperthermic process induced by 4 mg/kg of morphine.     

Involvement of nitric oxide in long-term potentiation of spinal nociceptive responses in rats

Nitric oxide plays an important role in spinal nociception. The present study explored the effects of nitric oxide on the spinal long-term potentiation associated with nociception. (1) Nitric oxide synthase inhibitor L-NAME (1 mM, 20 microl) and the nitric oxide scavenger hemoglobin (2 mg/ml, 20 mul) strikingly blocked the induction of tetanic sciatic stimulation-induced spinal long-term potentiation of C-fiber-evoked field potentials. L-arginine, a substrate of nitric oxide synthase, completely reversed L-NAME-induced inhibition. However, D-NAME (1 mM, 20 microl), an inactive form of L-NAME, had little effect on the spinal LTP. (2) The same tetanic sciatic stimulation induced long-term thermal hyperalgesia, which was blocked by pre-application of L-NAME. These results suggest the involvement of nitric oxide in the spinal long-term potentiation of C-fiber-evoked field potentials and related behavior changes.     

Basolateral amygdala lesions disrupt latent inhibitionin rats

Hypothermia is a well-known phenomenon which accompanies hypoglycemia in mammals. The present study was designed to test the hypothesis that nitric oxide (NO) plays a role in insulin-induced hypothermia. The body temperature (Tb) of awake, unrestrained rats was measured before and after systemic infusion of insulin (2U x kg(-1) x h(-1)), and intracerebroventricular administration of NG-nitro-(L)-arginine methyl ester (L-NAME, a nonselective NO synthase inhibitor, 200 microg/1 microl). We observed a significant reduction in body temperature after insulin infusion. L-NAME alone caused no significant change in body temperature. When the two treatments were combined, no change in Tb was observed. The data indicate that NO plays a key role in insulin-induced hypothermia.     

Effects of nitric oxide on passive avoidance learning in rats

To investigate the effects of nitric oxide (NO) on passive avoidance learning, L-NAME, D-NAME, and L-arginine were administered i.p. 30 min prior to learning trial; the effects of these substances were tested 24 h later using a passive avoidance apparatus in rats. To reveal the effect of NO on consolidation of acquired memory, L-NAME, D-NAME, and L-arginine were administered i.p. immediately after learning trials and animals were tested 24 h later. Effect of NO on retention was also investigated by injecting L-NAME, D-NAME, and L-arginine (same dosages) 30 min prior to 24 h testing (retrieval). L-NAME administered 30 min before and 24 h after learning trial significantly decreased the avoidance latency but there was no significant effect on consolidation. L-Arginine appeared to enhance the retention of acquired memory significantly, whereas D-NAME had no effect on any testing regime. The results suggest that NO may be involved in learning and retention of passive avoidance.     

A role for nitric oxide in the median eminence and arcuate nucleus response to capsaicin treatment in rats.

This study examined a possible functional involvement of nitric oxide (NO) in the median eminence (ME) and arcuate nucleus (ARC) after capsaicin treatment in rats. Subcutaneous injection of capsaicin increased nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) activity in the ARC-ME compared with vehicle treatment. Fos expression was increased in the ARC after capsaicin injection compared with vehicle-treated rats. Pretreatment with the NO synthase (NOS) inhibitor, N(omega)-nitro-L-arginine methyl ester (L-NAME) attenuated the effect of capsaicin on Fos expression and NADPH-d reactivity in the ARC-ME in comparison with rats injected with D-NAME, the inactive stereoisomer of L-NAME. These observations suggest that NO makes a major contribution to the response of the ARC-ME to a stressor such as capsaicin.     

The influence of L-NG-nitroarginine methyl ester, an inhibitor of nitric oxide synthase, upon the anticonvulsive activity of conventional antiepileptic drugs against maximal electroshock in mice

Summary. N^G-Nitro-L-arginine methyl ester (L-NAME, an unspecific nitric oxide synthase inhibitor), applied at 1 and 40 mg/kg, did not influence the electroconvulsive threshold, but impaired the anticonvulsant activity of valproate (at 40 mg/kg) and phenobarbital (at 1 and 40 mg/kg). No effect was observed in the case of carbamazepine and diphenylhydantoin. The effect of L-NAME upon the protective activity of phenobarbital was not reversed by L-arginine (500 mg/kg), a source of endogenous nitric oxide. Moreover, this nitric oxide synthase inhibitor did not alter the plasma levels of antiepileptic drugs studied, so a pharmacokinetic interaction is not probable. L-NAME per se (40 mg/kg) caused a moderate motor impairment but did not affect long-term memory. The combined treatment of L-NAME and antiepileptic drugs (providing a 50% protection against maximal electroshock) resulted in motor disturbances. On the other hand, mice performed the memory task better upon combined treatment of antiepileptic drugs with L-NAME compared to antiepileptic drugs alone. A 4-day administration of L-NAME, similarly to acute injections, decreased the protective action of phenobarbital but not that of diphenylhydantoin. The results indicate that L-NAME is able to reduce the protective activity of some conventional antiepileptics and this effect may be not associated with impaired synthesis of nitric oxide. Accepted November 19, 1997 / Received July 15, 1997     

Nitric oxide modulates articular sensory discharge and responsiveness to bradykinin in normal and arthritic rats in vivo

Nitric oxide is implicated in peripheral nociceptive processing. This study determined the effects of the nitric oxide synthase inhibitor, L-NAME, on neural discharge from articular C-fibre afferents innervating normal and arthritic ankle joints in anaesthetized rats. Intra-arterial injection of L-NAME (10-20 mg kg(-1)) increased neural discharge in normal and arthritic ankle joints, whereas D-NAME (30 mg kg(-1)) had no effect. The excitation induced by L-NAME (20 mg kg(-1)) was reduced by co-injecting the nitric oxide precursor, L-arginine (50 mg kg(-1)). L-NAME (20 mg kg(-1)) also enhanced responsiveness to bradykinin (10 microg) but only in arthritic rats, whereas L-arginine (50 mg kg(-1)) reduced the excitation by bradykinin (30 microg) in both groups. These results provide evidence that nitric oxide modulates articular C-fibre activity and reduces responsiveness to bradykinin.     

Nitric oxide synthase inhibition and MPTP-induced toxicity in the common marmoset

Nitric oxide, produced following activation of N-methyl-D-aspartate (NMDA) receptors, may be involved in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) toxicity since NMDA receptor antagonists have been shown to prevent MPTP induced nigral cell loss in primates. Common marmosets were treated with either saline or MPTP or L-N^Gnitro arginine methyl ester (L-NAME) or MPTP and L-NAME. MPTP-treated common marmosets showed motor deficits including bradykinesia, rigidity, and tremor accompanied by a marked loss of tyrosine hydroxylase-immunoreactive neurones in the substantia nigra pars compacta and of [³H]-mazindol binding in the caudate-putamen. MPTP treatment also caused an increase in glial fibrillary acidic protein (GFAP) staining in the substantia nigra compared to controls. However, MPTP treatment did not alter the number of constitutive nitric oxide synthase-immunoreactive neurones in the caudate-putamen. Furthermore, neurones or glial cells immunoreactive for inducible nitric oxide synthase were not observed in the substantia nigra pars compacta following MPTP treatment. L-NAME treatment alone did not produce any behavioural changes in marmosets and did not alter the number of tyrosine hydroxylase-immunoreactive cells in the substantia nigra pars compacta, the number of constitutive nitric oxide synthase-immunoreactive neurones or [³H]-mazindol binding in the caudate-putamen compared to saline-treated control animals. Furthermore, L-NAME did not affect the motor deficits, loss of tyrosine hydroxylase-immunoreactive neurones in the substantia nigra pars compacta, loss of [³H]-mazindol binding in the caudate-putamen, or the increase in GFAP staining in the substantia nigra induced by MPTP treatment of common marmosets. The failure of L-NAME to protect against MPTP-induced toxicity in the marmoset suggests that nitric oxide does not play a major role in such toxicity and casts doubt over the involvement of the NMDA:nitric oxide system in neurodegeneration in MPTP-treated primates. Synapse 26:301–316, 1997. © 1997 Wiley-Liss, Inc.     

EFFECTS OF NITRIC OXIDE ON PASSIVE AVOIDANCE LEARNING IN RATS

To investigate the effects of nitric oxide (NO) on passive avoidance learning, L-NAME, D-NAME, and L-arginine were administered i.p. 30 min prior to learning trial; the effects of these substances were tested 24 h later using a passive avoidance apparatus in rats. To reveal the effect of NO on consolidation of acquired memory, L-NAME, D-NAME, and L-arginine were administered i.p. immediately after learning trials and animals were tested 24 h later. Effect of NO on retention was also investigated by injecting L-NAME, D-NAME, and L-arginine (same dosages) 30 min prior to 24 h testing (retrieval). L-NAME administered 30 min before and 24 h after learning trial significantly decreased the avoidance latency but there was no significant effect on consolidation. L-Arginine appeared to enhance the retention of acquired memory significantly, whereas D-NAME had no effect on any testing regime. The results suggest that NO may be involved in learning and retention of passive avoidance     

Blockade of nitric oxide formation enhances thermal and behavioral responses in rats during turpentine abscess

OBJECTIVE: The purpose of this study was to investigate the role of nitric oxide (NO) during the development of fever and other symptoms of sickness behavior (i.e. anorexia, cachexia) in response to localized tissue inflammation caused by injection of turpentine in freely moving biotelemetered rats. METHODS: To determine the role of NO in turpentine-induced fever, we injected the NO synthase (NOS) inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME) intraperitoneally simultaneously or 5 h after turpentine injection. RESULTS: Rats responded with fever to intramuscular injection of 20 microl of turpentine that commenced 6 h after injection and reached peak values 11 h after injection. Although turpentine did not significantly alter food and water intake, it caused a drop in body weight. Rats injected with turpentine and treated with L-NAME responded with a substantial rise in fever, independently of the time of L-NAME injection. The rise in body temperature (T(b)) due to turpentine injection began slightly sooner and reached the maximal T(b) value faster in rats treated with L-NAME than in the ones treated with saline (control for L-NAME). The enhanced decrease in food and water intake in rats treated with a combination of L-NAME and turpentine was also observed. As a result, L-NAME-injected rats responded with a profound drop in body mass due to turpentine, independently of the time of L-NAME injection. L-NAME alone did not affect food and water intake, but slightly suppressed the gain of body mass. CONCLUSION: These results indirectly indicate that NO is involved in pyrogenic and behavioral responses in rats during turpentine abscess.