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There is a growing consensus that long-term deficits in the brain are due to dynamic interactions between multiple neural and immune cell types. Specifically, radiation induces an inflammatory response, including changes in neuromodulatory pro- and anti-inflammatory cytokine secretion. The purpose of this study was to establish that there is sympathetic involvement in radiation-induced decrements early in in vivo immune function host defense. Female, 8-9 week-old C57BL/6J mice were exposed to whole-body irradiation (WBI). There were 8 groups with radiation (0 vs. 3 Gy protons), immune challenge (Escherichia coli) and exposure to the sympathetic ganglionic blocker, chlorisondamine (1 mg/kg weight, i.p.), as independent variables. Ten days post-irradiation, mice were inoculated with E. coli intraperitoneally and sacrificed 90-120 min later. The data suggest that radiation-induced changes in immune function may in part be mediated by the sympathetic nervous system. Briefly, we found that radiation augments the bacteria-induced inflammatory cytokine response, particularly those cytokines involved in innate immunity. However, this augmentation can be reduced by the ganglionic blockade., (Copyright © 2015 European Federation of Immunological Societies. Published by Elsevier B.V. All rights reserved.)

CYP2B is a drug-metabolizing enzyme expressed in the liver and brain that metabolizes a variety of centrally acting drugs (e.g. propofol, bupropion and nicotine), endogenous neurochemicals (e.g. serotonin and testosterone) and toxins (e.g. chlorpyrifos). Human CYP2B6 is found at higher levels in the brains of smokers, and 7-day nicotine treatment induces rat brain CYP2B while not altering hepatic CYP2B. We characterized the time course of rat brain CYP2B induction by nicotine and determined if nicotinic acetylcholine receptors (nAChRs) mediated this induction. Rats were treated once daily with 1mg/kg nicotine base or saline s.c. for 1 or 7days and sacrificed from 30minutes to 7days after the last injection. One-day nicotine treatment did not induce brain CYP2B, whereas 7-day nicotine treatment significantly increased CYP2B expression for up to 24hours in the frontal cortex and brainstem; these levels returned to baseline by 7days post-treatment. CYP2B activity was also higher at 24hours in these regions. No change was seen in the cerebellar CYP2B levels or in vivo activity following nicotine treatment. Time of day of treatment and sacrifice altered the magnitude of brain CYP2B induction while chlorisondamine, an irreversible nAChR blocker, pre-treatment did not block CYP2B induction. Seven-day nicotine treatment resulted in an induction of rat brain CYP2B protein and in vivo activity for up to 24hours, which would suggest greater local drug metabolism by brain CYP2B. Humans or animals exposed to nicotine may have altered therapeutic drug response, brain levels of neurotransmitters and/or neurotoxicity., (Copyright 2010 Elsevier B.V. All rights reserved.)

We tested the hypothesis that in spontaneously hypertensive rat (SHR) NO produced centrally influences the resting arterial blood pressure by attenuating mechanisms involving prostaglandins, angiotensin II, endothelin and sympathetic nervous system. L-NAME (200 micro g/5 micro l), an inhibitor of NO synthase, administered intracerebroventricularly (i.c.v.) to awake and freely moving rats increased mean arterial blood pressure (MABP) in a biphasic pattern: an early transient increase within 1 min and a late prolonged response starting at 45 min and persisting for the duration of experiment (180 min). The two pressor responses involve different neurochemical mechanisms and, based on their latencies, they appear to reflect different anatomical sites of action of L-NAME. The late, but not the early pressor response, was prevented by pretreatment with chlorisondamine (2.5 mg/kg, i.v.), a ganglionic blocker, indicating its dependence on the sympathetic nervous system. Both pressor responses were abolished by i.c.v. pretreatment with indomethacin (200 micro g/5 micro l, i.c.v.), an inhibitor of cyclo-oxygenase, showing that they are mediated by prostaglandin(s). In contrast, losartan (25 micro g/5 micro l), an angiotensin II AT(1) receptor antagonist, had no effect. The initial pressor response was also attenuated by pretreatment with the endothelin ET(A)/ET(B) receptor antagonist, PD 145065 (48 micro g/2 micro l, i.c.v.). Intravenous pretreatment with another ET(A)/ET(B) receptor antagonist, L-754,142 (15 mg/kg as a bolus+15 mg/kg/h for 180 min), however, attenuated both responses to L-NAME. It is possible that L-754,142 crossed the blood-brain barrier and blocked, in addition, central ET(A)/ET(B) receptors. These studies show that NO synthesized in the brain attenuates pressor mechanisms involving prostaglandin, endothelin and sympathetic nervous system, but not angiotensin II, to modulate resting arterial blood pressure.

When exposed to extreme cold or injected with the alpha(2)-adrenoceptor agonist, clonidine, infant rats emit ultrasonic vocalizations (USVs). Based upon the cardiovascular changes that accompany these two manipulations, especially decreased venous return, it was hypothesized that USVs are the acoustic by-product of the abdominal compression reaction (ACR), a maneuver that increases venous return. If this hypothesis is correct, then other anithypertensive drugs that decrease venous return should evoke USVs. In Experiment 1, sodium nitroprusside (SNP, 400 microg/kg), a direct-acting dilator of arteries and veins, was administered to 15-day-old rats under thermoneutral conditions while cardiac rate and ultrasound production were monitored. In Experiment 2, femoral artery pressure was monitored after SNP administration. Infants responded to SNP administration with decreased arterial pressure and tachycardia and, in addition, significantly increased ultrasound production. In Experiment 3, chlorisondamine (5 mg/kg), a ganglionic blocker that causes vasodilation and bradycardia, and hydralazine (20 mg/kg), a selective dilator of arteries, was administered to 15-day-olds. As predicted, chlorisondamine evoked ultrasound production and hydralazine did not. These results introduce SNP and chlorisondamine as only the second and third known agents capable of independently evoking USVs in thermoneutral conditions, and provide further support for the notion that ultrasound production is triggered by decreased venous return.

Myoclonic twitching is a ubiquitous feature of infant behavior that has been used as an index of active sleep. Although the active sleep of infants differs in some ways from the REM sleep of adults, their marked similarities have led many to view them them as homologous behavioral states. Recently, however, this view has been challenged. One avenue for resolving this issue entails examination of the neural substrates of active sleep. If the neural substrates of active sleep were found to be similar to those of REM sleep, then this would support the view that the two states are homologous. Therefore, in the present study, decerebrations were performed in the pons and midbrain to determine whether the mesopontine region is important for the expression of active sleep in infants, just as it is for the expression of REM sleep in adults. It was found that, in comparison to controls, caudal pontine decerebrations reduced myoclonic twitching by 76%, rostral pontine decerebrations reduced twitching by 40%, and midbrain transections had no significant effect on twitching. Moreover, analysis of the temporal organization of twitching indicated that pontine decerebrations predominantly affected high-frequency twitching while leaving unaffected the low-frequency twitching that is thought to be contributed by local spinal circuits at this age. These results indicate that the mesopontine region plays a central role in the expression of active sleep in infant rats.

These experiments examined the effects of corticotropin-releasing hormone (CRH) on single-unit electrophysiological activity of locus coeruleus (LC) neurons. As has been reported previously, infusion of CRH into the ventricular system of the brain (i.c.v.) of halothane-anesthetized adult male rats increased spontaneous discharge rate of LC neurons while producing no increase, and possibly a decrease, in sensory-evoked activity. However, when i.c.v. CRH was given to female rats or immature male rats, which had not been studied previously, LC activity was not altered. To attempt to understand this sex and age difference, potential mechanisms by which i.c.v. CRH elevates LC spontaneous activity in adult male rats were examined; in that i.c.v. CRH activates the pituitary-adrenal axis and autonomic nervous system, these response systems were manipulated. Adrenalectomy (with or without corticosterone replacement by pellet) did not affect the ability of i.c.v. CRH to increase LC spontaneous activity in adult male animals, but blockade of sympathetically-mediated autonomic responses, either by chlorisondamine or the beta adrenergic receptor blocker timolol, blocked this increase, indicating that afferent feedback from peripheral autonomic responses was critical for activating LC neurons following i.c.v. CRH. To determine whether CRH neurotransmission might play a role in this feedback pathway, the CRH antagonist alpha-helical CRH (alpha-hCRH) was microinjected into several brain regions including LC prior to i.c.v. CRH. alpha-hCRH microinjected into LC reduced the increase in LC activity caused by i.c.v. CRH; however, blockade of this increase was total when alpha-hCRH was microinjected into the lateral parabrachial nucleus ipsilateral to the LC recording site, suggesting that increased LC activity following i.c.v. CRH is mediated by CRH acting in the parabrachial region. During these studies, it was also observed that microinjection of alpha-hCRH into LC increased LC spontaneous discharge rate; consequently, CRH was microinjected into LC, and produced a dose-dependent decrease in LC spontaneous activity in both male and female rats, which could be blocked by microinjection of alpha-hCRH - these data indicated that the direct influence of CRH on LC neurons is to decrease their spontaneous activity. To reconcile this with the original report that CRH applied to LC neurons increases their activity, one possibility suggested is that the CRH microinjection procedure used in the present study stimulated inhibitory receptors on LC dendrites whereas the original study stimulated excitatory receptors on LC cell bodies. It is concluded that an inhibitory influence of CRH on LC activity is consistent with recent data indicating that decreased LC activity increases anxiety and stress-related responses, but that direct influences of CRH appear rather minor in determining LC neuronal activity in comparison to other inputs to LC such as are seen after i.c.v. CRH infusion.

The 100 inescapable tail-shock paradigm produces three sequential analgesic states as the number of shocks increases: an early opioid analgesia (after 2 shocks) that is attenuated by systemic naltrexone, a middle analgesia (after 5-40 shocks) that is unaffected by systemic naltrexone, and a late opioid analgesia (after 80-100 shocks) that is attenuated by systemic naltrexone. In order to determine whether the absence of adrenal hormones would affect any of these analgesias, we tested adrenalectomized (ADX) versus sham-operated control rats 2 weeks post-surgery. Pain threshold was assessed using the tail-flick (TF) test. ADX attenuated both the early (2 shock) and late (80-100 shock) opiate analgesias and failed to reduce the naltrexone-insensitive analgesia after 5-40 shocks. We demonstrated that a loss of adrenomedullary catecholamines does not underlie the ADX-induced attenuation of opioid analgesia since sympathetic blockade using systemic chlorisondamine (6 mg/kg) failed to reduce analgesia at any point in the shock session. It was further shown that stress levels of adrenal hormones are not critical since (a) analgesia was unaffected when animals were tested 48 h after ADX, (b) 2 shocks do not produce a surge in corticosterone (CORT) over and above levels observed in animals restrained and TF tested in preparation for shock, and (c) basal CORT replacement in drinking water fully restored analgesia in ADX rats. These experiments demonstrate that basal CORT, rather than adrenomedullary substances, is critical to the expression of analgesia. The function of CORT here is not linked to a shock-induced surge of the steroid. CORT appears to play a permissive role in the expression of analgesia. Potential effects of the absence of corticosteroids on neurotransmitter biosynthesis important in analgesia production are discussed.

N-Methyl-D-aspartate (NMDA) microinjection (1 mM, 0.2 microliter) into the hypothalamic supraoptic nucleus (SON) stimulated heart rate in urethane-anaesthetized rats. This effect was inhibited by coinjection of a competitive blocker of NMDA receptors, CPP (20 nmol) or by pretreatment with a sympathetic ganglionic blocker, chlorisondamine chloride (5 mg/kg i.p.), but not by prior hypophysectomy. Furthermore, the cardioexcitatory effect of intra-SON NMDA was inhibited by prior intra-SON injection of a competitive blocker of nitric oxide (NO) synthesis, NG-nitro-L-arginine methyl ester (40 nmol) or a blocker of the soluble guanylate cyclase, Methylene blue (20 nmol), and was mimicked by intra-SON injection of a calcium ionophore, A23187 (10 nmol), which stimulates NO production by raising intracellular free calcium levels. Finally, intra-SON microinjection of a membrane-permeating cGMP analog, 8-bromo-cGMP (20 nmol) stimulated heart rate in urethane-anaesthetized rats. The results point to a functional link between a sympathetically mediated cardiophysiological effect of NMDA receptor stimulation in the SON and activation of the NO/cGMP signal transduction pathway.

Rats were trained to discriminate 1.9 mumol/kg (-)-nicotine (0.3 mg/kg) from saline in a standard two-bar operant conditioning paradigm with food reinforcement. The effect of neuronal nicotinic acetylcholine receptor (nAChR) agonists and antagonists were verified, and the participation of dopaminergic receptors subtypes in the expression of the (-)-nicotine cue was investigated with cis-flupentixol (D1-D2 antagonist), haloperidol (D2 antagonist) and clozapine (D4 antagonist). The stereoselectivity of the behavioral response was indicated by the 10-fold less sensitivity to (+)-nicotine in (-)-nicotine-trained rats. (+/-)-Anabasine and (-)-cytisine exhibited partial agonist profiles at the 1.9 mumol/kg dose while (-)-lobeline was devoid of any effect in doses up to 19 mumol/kg. (-)-Lobeline did not show antagonist properties in this paradigm. The nicotinic channel blockers mecamylamine, chlorisondamine and hexamethonium were inactive on their own but mecamylamine and chlorisondamine were able to block the effect of (-)-nicotine. Clozapine attenuated the (-)-nicotine cue while cis-flupentixol and haloperidol were ineffective. Similar doses of cis-flupentixol significantly blocked the locomotor stimulant effect of (-)-nicotine in rats indicating that blockade of dopaminergic receptors was achieved at the doses used in the drug discrimination studies. These data suggest that the discriminative stimulus properties of (-)-nicotine are mediated through neuronal nAChRs and involves the activation of dopaminergic receptors of the D4 subtype.

Intracerebroventricular (i.c.v.) angiotensin II (ANG II) increases vascular resistance and elicits a pressor response characterized by sympathetic nervous system activation (SNS component) and increased vasopressin (VP) secretion (VP component). This study examines the role of brain AT1 and AT2 ANG II receptors in mediating the pressor and renal hemodynamic effects of i.c.v. ANG II in conscious Sprague-Dawley rats. Mean arterial pressure, heart rate and renal vascular resistance responses to i.c.v. ANG II (100 ng in 5 microliters) were determined 10 min after i.c.v. injection of either the AT1 receptor antagonist, DuP 753 (1.0, 2.5, 5.0, 10.0 micrograms), the AT2 receptor ligand, PD 123319 (3.5 x [10(-6), 10(-4), 10(-2), 10(0)] micrograms), or both. In control rats, i.c.v. DuP 753 prevented the pressor response and the increase in renal vascular resistance that occurred following i.c.v. ANG II in a dose-dependent manner (P < 0.05), while i.c.v. PD 123319 was without affect. When the VP- and SNS components were studied individually, by preventing the SNS component with intravenous (i.v.) chlorisondamine or the VP component with a V1 receptor antagonist (i.v.) similar results were obtained; DuP 753 prevented the SNS component and significantly reduced the VP component. These results indicate that both central ANG II pressor components are mediated primarily by brain AT1 receptors. However, doses of DuP 753 were more effective when combined with 3.5 micrograms of PD 123319 than when given alone (P < 0.05), suggesting that the pressor effects of i.c.v. ANG II may involve activation of multiple ANG II receptor subtypes.

Glutamate microinjection (1 M, 250 nl) into the hypothalamic supraoptic nucleus (SON) stimulated heat production in brown adipose tissue (BAT) and caused a rapid and sustained increase in interscapular BAT and core temperatures in urethane-anaesthetized rats. This effect was blocked by intraperitoneal pretreatment with a sympathetic ganglionic blocker, chlorisondamine chloride (2.5 mg/kg), or a beta-adrenergic receptor blocker, propranolol (2.5 mg/kg), but not by prior hypophysectomy or intracerebroventricular pretreatment with specific receptor blockers to vasopressin (d(CH2)5[Tyr(Me)2]AVP, 5 micrograms) or oxytocin (d(CH2(5)[Tyr(Me)2,Thr4,Tyr-NH2(9)]OVT, 5 micrograms). The results demonstrate that stimulation of SON cells with glutamate elicits a non-vasopressinergic/non-oxytocinergic neural signal that can bring about a sympathetically-mediated increase in BAT thermogenesis. Heat production in BAT is an important mechanism of thermal protection during cold stimulation, and there is evidence that osmotic stimulation can influence thermoregulation. SON neurons play a major role in osmoregulation via release of the peptide hormones vasopressin and oxytocin. The present results suggest the possibility that apart from releasing peptide hormones for osmoregulation, SON neurons might be involved in mediating the effect of osmotic stimulation on thermoregulatory responses involved in thermal adaptation.

The posterior hypothalamus (PH) is involved in the generation of behavioral thermoregulatory responses, but the importance of the PH in the control of autonomic thermoregulatory responses such as heat production in brown adipose tissue (BAT) is not well defined. In the present study, selective stimulation of PH neurons by local application of the excitatory amino acid glutamate (250 nl of 1 M solution, unilaterally) caused a sharp, transient increase in interscapular BAT (IBAT) and core temperature in urethane-anesthetized rats. This effect was blocked by pretreatment with the sympathetic ganglionic blocker, chlorisondamine chloride (2 mg/kg) or the beta-adrenergic receptor blocker, propranolol (2 mg/kg), implicating the involvement of the sympathetic system. The effect of intra-PH injection of glutamate on IBAT and core temperatures could be mimicked by injection of the gamma-aminobutyric acid (GABA) receptor antagonist, bicucullin methiodide (BMI, 50 ng), into the same PH site. This effect of BMI could be blocked by co-injection of the GABA(A) receptor agonist, muscimol (25 ng). Further, BMI co-injection potentiated the effect of intra-PH injection of glutamate on IBAT and core temperatures. Conversely, muscimol co-injection prevented the stimulatory effect of intra-PH injection of glutamate. Taken together, the results indicate that direct chemical stimulation of neurons in the PH can activate an autonomic mechanism controlling heat production in BAT. Further, they suggest the neural mechanism in the PH mediating this effect is tonically inhibited by GABA, as blockade of GABAergic function in the PH produces an effect similar to that observed after direct stimulation of PH neurons with glutamate.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of intravenous (i.v.) serotonin (5-HT) on nociception and blood pressure was examined in male Sprague-Dawley rats. Intravenous 5-HT produced a dose-dependent (6-192 micrograms/kg, i.v.) inhibition of the nociceptive tail-flick (TF) reflex in lightly pentobarbital-anesthetized (ED50 = 40 micrograms/kg) and conscious rats (ED50 = 44 micrograms/kg). In the lightly pentobarbital-anesthetized rat, the blood pressure response to i.v. 5-HT was typically a triphasic response with a marked Bezold-Jarisch reflex-induced decrease in pressure (associated with a brief period of apnea) followed by a pressor phase and a subsequent delayed hypotension. In the conscious rat, the response was typically biphasic with the late hypotensive phase absent. A variety of anatomical and pharmacological manipulations were performed to characterize the 5-HT-induced inhibition of the TF reflex and associated changes in blood pressure. Prevention of 5-HT-induced reflex apnea by artificial ventilation did not affect inhibition of the TF reflex produced by 5-HT. Pharmacological manipulations were performed to mimic, as closely as possible, the acute increases and decreases in blood pressure associated with i.v. 5-HT. Nitroprusside (8 micrograms/kg, i.v.) produced a decrease in blood pressure of similar magnitude and rate as that associated with the Bezold-Jarisch reflex-induced decrease in pressure produced by 72 micrograms/kg 5-HT, but did not change TF latency from baseline. Similarly, acute increases in pressure produced by phenylephrine (8 micrograms/kg, i.v.), intended to mimic the secondary pressor effect of 5-HT, did not change TF latency. The short-acting ganglion blocker trimethaphan (5 mg/kg, i.v.) closely mimicked the late hypotensive phase produced by 5-HT, but again resulted in no change in TF latency. Pretreatment with the ganglion blocker chlorisondamine (2.5 mg/kg) abolished all depressor responses to 72 micrograms/kg 5-HT, but did not significantly affect the TF reflex. These results indicate that acute changes in blood pressure and respiration associated with i.v. 5-HT do not contribute to inhibition of the TF reflex. This conclusion was confirmed in experiments in which bilateral vagotomy abolished approximately 70% of the 5-HT-induced inhibition of the TF reflex (and all depressor responses), and resulted in a significantly greater pressor response. Finally, low thoracic spinal cord transection (T9-10) abolished the inhibition of the TF reflex produced by i.v. 5-HT. Therefore, 5-HT stimulates vagal afferents and inhibits the TF reflex by activating descending inhibitory systems from the brainstem.(ABSTRACT TRUNCATED AT 400 WORDS)

The ventromedial hypothalamic nucleus (VMH) has been recognized for its role in the control of thermogenesis in brown adipose tissue (BAT) in the rat. However, the neural elements within the VMH that might be involved have not been clearly identified. In the present study, intra-VMH microinjections of the excitatory amino acid glutamate (100 mM to 1 M, in 0.25 microliters), which excites cell bodies and dendrites but not axons and nerve terminals, dose-dependently increased interscapular BAT (IBAT) temperature in urethane-anaesthetized rats. This effect of glutamate was blocked by prior treatment with the sympathetic ganglionic blocker, chlorisondamine chloride (2.5 mg/kg, i.p.) or the beta-adrenergic receptor antagonist, propranolol (2.5 mg/kg, i.p.), implicating the involvement of sympathetic norepinephrine. These results are consistent with the view that cells in the VMH are implicated in the transmission of thermogenic signals to BAT.

The paraventricular nucleus (PVN) of the hypothalamus is involved in the control of energy balance in rodents through its influences on feeding, pituitary hormone secretion and the autonomic nervous system. In the present study, selective stimulation of PVN neurons by means of local microinjection of the excitatory amino acid glutamate (100 mM, 500 mM or 100 nl) led to a concentration-dependent increase in interscapular brown adipose tissue (IBAT) temperature in urethane-anaesthetized rats. This effect could be prevented by pretreatment with the sympathetic ganglionic blocker, chlorisondamine chloride, or the beta-adrenergic receptor antagonist, propranolol, but not by hypophysectomy, implicating the involvement of the sympathetic outflow. Thermogenesis in brown adipose tissue (BAT) is an important accompaniment of overfeeding in small mammals, and previous studies have shown that signals generated in response to feeding gain access to the PVN. The present finding that direct activation of PVN neurons stimulates thermogenesis in BAT, taken together with data that the PVN receives dietary signals from the gut, support the view that PVN neurons may function in monitoring the balance between energy intake and its expenditure.

This study was designed to compare the cardiovascular influences of the rostral ventrolateral medulla (RVLM) and the caudal ventrolateral medulla (CVLM) in young (5-6 weeks) spontaneously hypertensive (SH) and normotensive Wistar-Kyoto (WKY) rats. SH and WKY groups had similar pressor and depressor responses to microinjection of L-glutamate into the RVLM and the CVLM, respectively. In addition the results of this study indicate a reduced tonic sympathoinhibitory function of the CVLM in young SH rats, which may contribute to the development of hypertension in the spontaneously hypertensive rat.

Factors regulating the putative peptide neurotransmitter, substance P, were examined in the adult rat sympathetic superior cervical ganglion (SCG). Surgical section of the preganglionic nerves resulted in a 55% increase in ganglion content of SP-like immunoreactivity (subsequently termed SP). Treatment with chlorisondamine, which blocks nicotinic ganglionic transmission, also increased SP, reproducing the effects of decentralization. Conversely, treatment with phenoxybenzamine, which reflexly increases sympathetic activity, reduced ganglion SP. Axotomy of the postganglionic nerves did not alter ganglion SP. Finally, treatment of rats as neonates with 6-hydroxydopamine, which selectively destroys principal ganglion neurons, profoundly decreased SP in the adult SCG. Our observations suggest that impulse activity of preganglionic nerves decreases ganglion SP in principal ganglion neurons through a transsynaptic process. Viewed in conjunction with previous work, our studies suggest that transsynaptic stimulation has opposite effects on SP and noradrenergic transmitter characters in the SCG.

The role of normal synaptic activity in the biochemical development of the nervous system has been examined in the chick embryo. Chlorisondamine, a ganglionic blocking drug, was administered in ovo during the period of synaptogenesis in the parasympathetic ciliary ganglion. Following treatment with chlorisondamine, nicotinic binding sites (as measured with [125I] alpha-bungarotoxin) were significantly reduced in both the ganglion and its end organ, the striated iris muscle. While the number of [125I] alpha-bungarotoxin binding sites eventually approached control levels in the iris, binding in the ciliary ganglion remained below normal values through hatching.

Intravenous naloxone, 1 or 10 mg/kg, protects sensitized mice from lethal anaphylaxis. The protective effect is reversed by pretreatment with the ganglionic blocker, chlorisondamine chloride, peripheral chemical sympathectomy with 6-hydroxydopamine or bilateral adrenal gland denervation. The possible involvement of the sympathetic nervous system in naloxone's antianaphylactic action, suggested by these findings, is discussed.

Glycinergic mechanisms have been implicated in central cardiovascular regulation. However, the inhibitory amino acid's role in the nucleus tractus solitarii (NTS), the site of termination of cardiovascular afferents, has not been clarified. Thus, we sought to determine if the microinjection of glycine into the NTS alters arterial pressure and heart rate. Microinjections of glycine, like glutamate, confined to the NTS decreased arterial pressure and heart rate in a neurally mediated, dose-dependent manner. The glycine antagonist strychnine completely blocked these effects of glycine but did not itself alter arterial pressure or heart rate, or interfere with the baroreceptor reflex. The acute hypotensive, bradycardic response to glycine was followed by a period during which glycine essentially eliminated the cardiovascular responses to the microinjection into the NTS of glutamate, an amino acid reputed to be a transmitter in the baroreceptor reflex arc. These data suggest that glycine is involved in cardiovascular regulation by the NTS but do not support its being an integral transmitter in the baroreceptor reflex.

At the crayfish neuromuscular junction chlorisondamine reduced the amplitude of both the excitatory junctional potential and the glutamate current in a dose-dependent manner in concentrations above 3 microM, and it is suggested that the drug is a powerful non-competitive antagonist for glutamate. Chlorisondamine did not act presynaptically on the crayfish neuromuscular junction. A double exponential decay of excitatory synaptic currents was observed in the presence of chlorisondamine, suggesting that this drug is an open channel blocker for the excitatory neurotransmitter. The glutamate current tail was prolonged in the presence of chlorisondamine. This prolongation increased with increasing iontophoretic current of glutamate. The rate of recovery from the refractory form of the glutamate receptor to the free reactive one was hardly affected by chlorisondamine. The inhibitory action of chlorisondamine on glutamate responses was voltage-dependent and hyperpolarization reduced the drug action. Chlorisondamine depressed the glutamate current even in Na-free, Ca-rich solution.

Using the rat superior cervical ganglion in vivo as a model of the central nervous system, we have sought to determine whether postsynaptic elements as well as terminal processes are metabolically activated during impulse activity. The rate of glucose utilization in the ganglion was found to be stimulated by antidromic stimulation of the external carotid nerve. This stimulation was frequency-dependent and was confined only to the region in the caudal portion of the ganglion corresponding to the location of the cell bodies of origin of the external carotid nerve.

Unlabelled: Corticotropin releasing factor (CRF) given intracerebroventricularly (i.c.v.) increases mean arterial pressure (MAP) and heart rate (HR), while CRF given intravenously decreases MAP and increases HR. CRF given i.c.v. elevates plasma concentrations of vasopressin and catecholamines. Ganglionic blockade with chlorisondamine prevents CRF-induced increases in MAP; the vasopressin antagonist, [1-deaminopenicillamine,2-(O-methyl)tyrosine]-vasopressin does not alter CRF-induced increases in MAP. In contrast to CRF, angiotensin II (A-II) given i.c.v. increases MAP but decreases HR., In Conclusion: (1) CRF elevation of MAP and HR in dogs is dependent on an intact sympathetic nervous system, and (2) CRF and A-II have different CNS effects on cardiovascular function.

Exogenously applied opioid agonists have a stimulatory effect on adrenocorticotropic hormone (ACTH) secretion. The present experiments were designed to examine the mechanisms involved in the stimulatory effect of the mu-receptor agonist morphine on ACTH release in chronically cannulated, freely moving, non-stressed rats. Morphine (7.5 mg/kg, i.v.) treatment was followed by a significant increase in plasma levels of ACTH. Pretreatment with the peripheral ganglionic blocker chlorisondamine (3 mg/kg, i.p.) attenuated the response to morphine. The morphine stimulatory effect was also partially inhibited if the rats were pretreated with a specific antiserum to corticotropin-releasing factor (CRF). In rats given both CRF antiserum and chlorisondamine, the plasma ACTH levels remained unchanged after morphine application. These findings indicate that morphine stimulates the release of ACTH by activating both CRF-secretion and peripheral sympathetic neuronal pathways.

A single dose of reserpine administered into the yolk sac of chicken eggs prior to incubation produces two distinct periods of significant increase in tyrosine hydroxylase (TH) activity over controls. The first period is 21 days of incubation (55%) and the second is between day 14 and 30 after hatching (a.h.) (69%). Cholineacetyltransferase (ChAc) and dopadecarboxylase (DDC) are not modified in the two periods of increased TH activity. Reserpine had no effect on cholinergic parasympathetic synapses and neurons in the ciliary ganglion, as judged by ChAc activity. When reserpine was acutely administered in three different posthatching periods only the injection at the latest period (days 26 and 27) caused a significant (38%) increase in TH activity at day 30. Postsynaptic nicotinic receptors were blocked selectively by injecting chlorisondamine in the chick starting at hatching for one week. The administration of chlorisondamine almost completely abolished the reserpine induced increase of TH activity at day 15 a.h. The present results support the view that the development of enzyme activities specifically related to neurotransmitter biosynthesis in chick autonomic ganglia is regulated not only by transsynaptic influences but also by regulatory inputs originating in the periphery.

The cardiovascular and hormonal responses to intracerebroventricular administration of corticotropin-releasing factor (CRF) and angiotensin II (AII) were compared in conscious, freely moving rats. CRF elevated mean arterial pressure (MAP) and heart rate (HR), increased plasma catecholamine levels and had no effect on plasma vasopressin levels. AII increased MAP and decreased HR, had no effect on plasma catecholamine levels and elevated plasma vasopressin levels. Ganglionic blockade with chlorisondamine blocked the cardiovascular effects of CRF but failed to antagonize the pressor response to AII. The vasopressin antagonist, [1-deaminopenicillamine, 2-(0-methyl)tyrosine]-AVP, did not modify CRF-induced cardiovascular changes but greatly attenuated AII's effects on blood pressure. These results suggest that CRF and AII have different central actions on hormonal and cardiovascular systems.

The ganglionic blocking agents, chlorisondamine (CL) and hexamethonium (HEX) were used to examine the role of altered autonomic function in the behavioral response to i.c.v.-administered corticotropin-releasing factor (CRF). Animals were tested in either a novel modified open field or in their home cages. CRF-induced alterations in locomotion, grooming and eating were assessed in both environments in the presence or absence of CL or HEX. In the home cage the ability of CRF to increase grooming was attenuated by pretreatment with either CL or HEX. In the modified open field only HEX significantly suppressed grooming. In the familiar environment CRF-induced increases in locomotion were significantly inhibited by CL. However, in a novel environment, where CRF suppresses locomotion, CL was inactive. The competitive ganglionic nicotinic blocking agent, HEX, on the other hand, inhibited both the increased locomotion produced by CRF in the home cage and also the decreased locomotion induced by CRF in the modified open field. CRF suppression of food consumption was attenuated by CL. These results indicate that while centrally-mediated activation of the sympathetic nervous system cannot account for the full magnitude of the behavioral effects of i.c.v. CRF, such activation may play a part in both the locomotor activating components of the CRF response seen in the familiar environment as well as the suppressive effects seen in the novel environment.

The role of nicotinic cholinergic transmission in cold stress-induced alterations in rat adrenomedullary tyrosine hydroxylase (TH) mRNA was investigated by RNA dot-blot hybridization, using a cloned TH cDNA probe. Chlorisondamine, a ganglionic blocking agent, greatly attenuated the induction of TH mRNA levels caused by cold exposure, whereas carbachol and nicotine, cholinergic agonists, increased TH mRNA in control animals. These results suggest that cholinergic nicotinic receptors play a key role in the transsynaptic induction of adrenal TH gene expression.

gamma-Endorphin-type peptides (i.e. gamma-endorphin, des-tyr'-gamma-endorphin [DT gamma E]) result from the processing of the opioid peptide, beta-endorphin. Previous studies have implicated the involvement of beta-endorphin in various types of shock, including anaphylactic shock. In the present experiments the intracerebroventricular (i.c.v.) administration of gamma-endorphin (10 micrograms) or DT gamma E (3.3-10 micrograms) significantly improved survival in anaphylactic shock in mice. Moreover, DT gamma E (10 micrograms) reversed the effect of i.c.v. beta-endorphin (3.3 micrograms) to exacerbate shock. A similar dose of DT gamma E was ineffective in antagonizing beta-endorphin-induced analgesia. The anti-anaphylactic action of DT gamma E as well as its effect to block the pro-anaphylactic action of beta-endorphin were prevented by pretreatment with the sympathetic ganglionic blocker, chlorisondamine chloride. The results suggest that gamma-endorphin-type peptides may act in the central nervous system (CNS) to physiologically oppose the autonomic pathophysiologic influences of beta-endorphin.