CCK4-induced panic in healthy subjects I: psychological and cardiovascular effects.

Sixteen healthy subjects participated in a crossover, double blind, and placebo-controlled study, designed to assess simultaneously the psychological and cardiovascular effects of cholecystokinin tetrapeptide (CCK4). Following an i.v. injection of 25 microg of CCK4, 44 percent of subjects experienced symptoms that fulfilled the DSM-IV criteria for a panic attack while no one panicked with placebo. CCK4 induced a significantly greater number and higher intensity of panic-like symptoms than placebo. A significant increase in state anxiety was observed in the period after CCK4 injection; this increase was significantly larger than the non-specific anxious reaction to placebo. CCK4 also affected cardiovascular signs. Both heart rate and mean blood pressure significantly increased after administration of CCK4. Again, these increases were significantly higher than those seen after placebo injection. We conclude that, in healthy subjects, CCK4 induces panic-like reaction characterized by a number of somatic, cognitive and emotional symptoms, which are accompanied by increases in heart rate and blood pressure.

CCK4-induced panic in healthy subjects II: neurochemical correlates.

Cholecystokinin tetrapeptide (CCK4) induces symptoms similar to those of panic attack. The present study investigated the effects of CCK4 administration on catecholaminergic system. In this double blind, randomised, crossover experiment, 16 healthy subjects received injections of either 25 microg of CCK4 or placebo on two separate occasions. Platelet and plasma catecholamine concentrations were assessed before the administration and compared to post-injection values. The results clearly show that both plasma and platelet concentrations of catecholamines are significantly affected by CCK4. Plasma norepinephrine (NE) and epinephrine (EPI) raised significantly above baseline in the immediate post-CCK4 period, while in plasma dopamine (DA), the significant increases were delayed. In the platelets, significant post-CCK4 increases of NE and EPI concentrations were observed with a delay of several minutes. In summary, we have demonstrated that, in healthy subjects, CCK4 increases peripheral concentrations of catecholamines in both plasma and platelets, with the most consistent changes occurring in platelet NE and plasma EPI concentrations.

Elevated plasma levels of neuropeptide Y in patients with panic disorder.

OBJECTIVE: Neuropeptide Y is a pancreatic polypeptide closely associated with noradrenergic activity both in the central and peripheral nervous systems. The objective of this study was to assess plasma neuropeptide Y-like immunoreactivity in panic disorder. METHOD: Radioimmunoassays were performed in 12 patients with DSM-III-R panic disorder and two groups of normal comparison subjects (N = 22 and N = 16). RESULTS: Markedly higher plasma neuropeptide Y-like immunoreactivity was found in patients with panic disorder. CONCLUSIONS: Higher plasma neuropeptide Y-like immunoreactivity suggests that this peptide may be implicated in the etiology or expression of symptoms of panic disorder.

Tolerance to the antinociceptive properties of morphine in the rat spinal cord: alteration of calcitonin gene-related peptide-like immunostaining and receptor binding sites.

Tolerance to the spinal antinociceptive effects of morphine develops rapidly after its chronic administration. The mechanism involved in this phenomenon is unclear, but it is unlikely due to a direct regulation of spinal opioid peptides and their receptor binding sites. A variety of neuropeptides, especially the neurokinins and calcitonin gene-related peptide (CGRP) are concentrated in primary sensory afferents and have thus been proposed to play significant roles in spinal nociceptive mechanisms. However, their functions in the development of tolerance to the antinociceptive properties of morphine have not been explored fully. We therefore investigated the possible involvement of various sensory neuropeptides including CGRP, substance P, galanin, neurotensin and neuropeptide Y and their receptors in the dorsal horn of the spinal cord during the development of tolerance to the antinociceptive action of intrathecal morphine. Morphine sulfate (7.5 micrograms/microliters/hr) was administered continuously at lumbar level L4 using mini-osmotic pumps for 3, 5, 7 and 14 days. Tolerance to the antinociceptive effect of morphine was verified with the tail-immersion test and became evident on the 5th day of treatment. In tolerant animals, there was a marked increase in CGRP-like immunostaining and a decrease (30-45%) in [125I]human CGRP alpha binding in laminae I, II and III of the dorsal horn of the spinal cord. These changes coincided with the onset of morphine tolerance and persisted for the 14-day period during which tolerance was present. Similar changes were not observed in the immunostaining or binding of the other neuropeptides studied.(ABSTRACT TRUNCATED AT 250 WORDS)

In vivo central actions of rat amylin.

The purpose of the present study was to examine and compare the profile of neurobehavioral effects of rat amylin (r-amylin) and rat calcitonin gene-related peptide (rCGRP), two peptides having a 50% structural homology. The effects of synthetic r-amylin and rCGRP administered in several doses (0.312-80.0 micrograms) into the lateral cerebro-ventricle of rats on spontaneous activity, muscular tone, body temperature, nociception, food intake as well as their potential for inducing catalepsy, were investigated. Intraventricular administration of r-amylin or rCGRP significantly reduced spontaneous motor activity and markedly increased body temperature of animals in a dose-dependent related fashion. rCGRP produced a significant increase in muscular tone and induced cataleptic effect in animals, but r-amylin had no effect on these variables. Furthermore, neither r-amylin nor rCGRP were able to induce any significant effect on nociceptive response time of animals in the tail immersion test even with doses as large as 80.0 micrograms. Finally, the two peptides did not affect ad libitum food intake, but significantly reduced food consumption in 22 h food-deprived animals. Together, the results of the present study suggest that amylin may be involved in a diversity of neurophysiological processes but displays a different profile of neurobehavioral effects to that of CGRP which may involve different receptors.

Influence of ambient temperature on the effects of NPY on body temperature and food intake.

Because thermoregulation and food consumption are interrelated, and because thermoregulation processes are influenced by ambient temperature, we examined the effects of neuropeptide Y (NPY) on both body temperature and food intake in various thermal environments after intracerebroventricular administration of 20 micrograms. Results reveal that the prominent effects of NPY on body temperature and food intake in relatively thermoneutral environments are drastically altered at more extreme ambient temperatures. NPY produced hypothermia in animals placed at 4, 12, and 21 degrees C, and actually increased body temperature in animals subjected to 30 and 38 degrees C temperature. On the other hand, in comparison with ambient temperatures of 12 and 21 degrees C, ambient temperatures of 4 and 30 degrees C significantly reduced the stimulatory effect of NPY on food consumption. Moreover, at 38 degrees C the effect of NPY on food intake was totally abolished. These data demonstrate that ambient temperature has a critical influence on central actions of NPY.

Differential effects of low versus high doses of apomorphine on retinal dopamine metabolism in light- and dark-adapted rabbits.

Previous electrophysiologic results from this laboratory indicate that apomorphine exerts a differential dose-related effect on rabbit electroretinograms, with low doses increasing the b-wave and higher doses decreasing this parameter. Results were interpreted as reflecting apomorphine's agonistic properties at two different receptors: 1.0 mg/kg acting at the postsynaptic site, and the lower dose, 0.01 mg/kg, preferentially stimulating inhibitory autoreceptors. The purpose of this experiment was to investigate further this hypothesis by determining retinal levels of dopamine, dihydroxyphenylacetic acid, and homovanillic acid in retinas of light- or dark-adapted rabbits treated with saline, 1.0, 0.1, or 0.01 mg/kg apomorphine intravenously. Results indicate that in dark-adapted rabbits only the highest dose tested, 1.0 mg/kg, decreased dopamine concentrations. In animals exposed to light, the lowest dose tested, 0.01 mg/kg, significantly reduced dopamine and metabolite levels, whereas the highest dose unexpectedly increased retinal dopamine turnover. Results are discussed in terms of receptor sites and the influence of lighting conditions.

Mapping of hypothalamic sites involved in the effects of NPY on body temperature and food intake.

The objective of the present study was to identify hypothalamic sites that might be implicated in the effects of neuropeptide Y (NPY) on both body temperature and food intake. For this purpose, the effects of direct microinjections of NPY in several doses (0.156-20 micrograms) into discrete hypothalamic nuclei on body temperature were examined in rats. To examine specificity of effects, food consumption of animals following injections was also measured. Results indicate that the influence of NPY on body temperature varies with the hypothalamic region where the peptide is administered. NPY had no effect on temperature after administration into the ventromedial (VMH) and the perifornical hypothalamus (PeF). However, a significant hypothermia was seen following administration into the preoptic (POA) and arcuate nucleus (Arc), and hyperthermia was seen after injection into the paraventricular nucleus (PVN). Finally, a biphasic effect was observed after injection into the lateral hypothalamus (LH): hyperthermia with relatively small doses and hypothermia with higher doses. Similar effects were obtained when administered into the third ventricle (3V) but in an inverted dose-related fashion: hypothermia at low and hyperthermia at higher doses. For feeding, NPY consistently increased food intake in all regions examined, with the strongest effect obtained after administration into the PeF. The present results clearly dissociate the effects of NPY on food intake and body temperature, and demonstrate that these effects are related to specific hypothalamic nuclei.

Effects of NPY and NPY2-36 on body temperature and food intake following administration into hypothalamic nuclei.

Our previous in vivo structure-activity studies suggested that the putative receptors mediating the effects of NPY and NPY2-36 on food intake and body temperature are pharmacologically different [17]. In the present study, we examined and compared dose-related effects of NPY and NPY2-36 on ad lib food intake and rectal temperature after administration into discrete hypothalamic nuclei of the rat. Results indicate that NPY and NPY2-36 have opposite effects on body temperature to those of NPY when injected in the preoptic area (POA): hypothermia and hyperthermia, respectively. When administered in the paraventricular nucleus (PVN), both increased body temperature. When injected into the third ventricle (3V), NPY produced a biphasic effect: hypothermia at low doses and hyperthermia at high doses. Similar effects were obtained with NPY2-36, but in an inverted dose-related fashion: hyperthermia at low and hypothermia at higher doses. In the arcuate nucleus (Arc), NPY induced a significant hypothermia whereas NPY2-36 had no effect. Finally, neither peptide affected body temperature when injected into the ventromedial (VMH) and perifornical (PeF) nuclei. Both NPY and NPY2-36 increased food intake after injection in all regions examined. In general, NPY was more potent and efficacious than NPY2-36. The present results clearly dissociate the effects of NPY on food intake and body temperature. Furthermore, the data support the hypothesis that the putative receptors underlying the effects of NPY and NPY2-36 on food intake are similar, whereas those mediating the effects on body temperature are pharmacologically different.

Structure-activity analysis of the motor effects of neuropeptide Y.

The purpose of the present study was to examine structure-activity relationships for three known motor effects of neuropeptide Y (NPY): decreased spontaneous activity, enhancement of muscle tone, and catalepsy. Various NPY fragments and structural analogues were synthesized and administered intracerebroventricularly in several doses (2.5-160 micrograms); their effects on these three motor variables were evaluated and compared. Globally, results indicate that the C-terminal portion of the peptide is responsible for the three motor effects of NPY. However, the distributions of potencies of the various fragments and analogues for each effect were clearly different, suggesting that the putative receptors mediating each motor effect are pharmacologically different. The findings of the present work are discussed in relation to those obtained in previous structure-activity studies.

Differential effects of pertussis toxin on body temperature changes induced by neuropeptide Y and NPY2-36.

Many effects of NPY have been attributed to a decrease in the activity of adenylate cyclase. Pre-treatment with pertussis toxin (PTx) has been shown to inhibit many pharmacological effects of NPY including increased feeding following administration in the paraventricular nucleus (PVN). In the present study, we examined the influence of PTx pretreatment on the effects of NPY on body temperature following administration in the preoptic area (POA), a region which seems to be the most sensitive to the effects of the peptide on body temperature. The effects of the same pre-treatment on the action of NPY2-36 was also studied since we have found previously that this fragment produced opposite effects on body temperature to that of NPY when injected in the POA. PTx was administered 3 days prior to the injection of NPY or NPY2-36. Results indicate that the hypothermic effect of NPY produced in the POA was blocked by PTx whereas the hyperthermic effect of NPY2-36 was not affected. These results are important as they provide evidence that, in the POA at least, the receptors mediating the hypothermic effect of NPY might be biochemically different from those mediating the hyperthermic effect of NPY2-36.

Neurochemical effects of neuropeptide Y (NPY) and NPY2-36.

Our previous in vivo structure-activity studies suggested that the putative receptors mediating the effects of NPY and NPY2-36 on food intake and body temperature following ICV administration are pharmacologically different. In the present study, we examined and compared dose related effects of NPY and NPY2-36 on levels of norepinephrine (NE), dopamine (DA) and its main metabolites, dihydroxyphenylacetic acid (DOPAC) and homovanilic acid (HVA), as well as serotonin (5-HT) and its metabolite, 5-hydroxyindolacetic acid (5-HIAA), in several brain regions of the rat, including: frontal cortex, hypothalamus, amygdala, septum, nucleus accumbens, corpus striatum, globus pallidus, substantia nigra and hippocampus. NPY and NPY2-36 (10 or 20 micrograms) were administered intraventricularly and the regional levels of the amines and metabolites were assessed 30 min following administration. Results indicate that both doses of NPY decreased NE levels within the hypothalamus. Furthermore, DOPAC concentrations were increased in this region while DA and HVA remained unchanged. The most pronounced neurochemical effects of NPY were found in the hippocampus, where the peptide produced dose related increases in DA, DOPAC and HVA. On the other hand, NPY2-36 significantly increased NE, DA and its metabolite DOPAC in both the amygdala and septum. The metabolism of DA was most obviously affected in the hippocampus and frontal cortex where levels of DA and DOPAC were significantly increased. 5-HT was affected in both the hypothalamus and globus pallidus where DA and its metabolite HVA were also increased.(ABSTRACT TRUNCATED AT 250 WORDS)

Agonist and antagonist opioid activity of axial and equatorial conformations of S-methyl- and S-allyl-morphinans.

Resolved axial (beta) and equatorial (alpha) forms of S-methyl (beta-sulforphanol, alpha-sulforphanol) and S-allyl (beta-sulfallorphan, alpha-sulfallorphan) morphinans were tested for their ability to depress the electrically evoked contractions of the guinea pig ileum and of the mouse vas deferens, to compete with the binding of prototype ligands selective for mu-, delta-, and kappa-opioid receptors in membrane preparations of rat brain and guinea pig cerebellum and to produce analgesia in a rat thermal pain assay. beta-Sulforphanol was more potent than alpha-sulforphanol in the guinea pig ileum (relative potencies of 93% and 29% respectively, as compared with levorphanol). beta-Sulfallorphan and alpha-sulfallorphan were both inactive in the guinea pig ileum assay. In the mouse vas deferens preparation, beta-sulforphanol and alpha-sulforphanol had relative potencies of 2.1% and 1.2% as compared with levorphanol, respectively, while the S-allyl derivatives were inactive. All morphinan derivatives displayed marked binding selectivity for mu-opioid receptors but alpha-sulfallorphan also showed significant binding potency on delta-opioid receptors (12% as compared to levorphanol). The compounds were also tested for their ability to antagonize the biological activity of morphine. In the guinea pig ileum, alpha-sulfallorphan potently inhibited morphine with a Ke value of 41.7 nM. alpha-Sulforphanol also antagonized morphine but with a smaller potency (Ke = 350 nM). In the mouse vas deferens, no antagonist activity against morphine was observed with any morphinan derivative tested at 1 microM. In the rat thermal pain assay, beta-sulforphanol (intracisternally, i.c.) was more potent than alpha-sulforphanol in producing analgesia while the other morphinan derivatives were inactive.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of dextromethorphan on ischemia induced electroretinogram changes in rabbit.

The present study was undertaken to determine whether dextromethorphan (DM), a potent N-methyl-D-aspartate antagonist, could attenuate the effects of ischemia on rabbit ERG. Retinal ischemia was induced by increasing intraocular pressure to 120 mm Hg for 30, 60, or 90 min. DM was intravenously administered before ischemia and maintained throughout the entire experimental period. ERGs were recorded prior to, during, and after ischemia. The results indicate that the b-wave hyperresponses and the delays in implicit times induced by 30 min. ischemia were suppressed by the administration of DM. Similar findings were obtained when ischemia lasted for 60 min, except that DM did not improve delayed implicit times, suggesting that cellular injury is still present. ERG changes resulting from 90 min ischemia were not reversed by DM treatment. Effects of DM treatment on a-wave were less prominent. Together, our results further support that DM can to some extent alleviate ischemic injury in the rabbit retina.

Selective effects of retinal dopamine depletion on partial ischemia-induced electroretinographic hyperresponses in rabbits.

The interaction of retinal dopamine depletion and partial ischemia on the a- and b-wave amplitudes and implicit times of the electroretinogram was examined in adult pigmented rabbits. Seven days after 6-hydroxydopamine treatment, which resulted in a depletion of the amine, partial retinal ischemia was induced by raising the intraocular pressure. As expected, moderate elevation of intraocular pressure produced increases in both a- and b-wave amplitudes. Amplitude hyperresponses were significantly reduced in dopamine-depleted retinas. These reductions were more prominent with relatively lower intensities. However, response delays were not shortened but lengthened by 6-hydroxydopamine pretreatment. Together, these results point to a selective role of dopamine in partial retinal ischemia induced by moderate elevation of intraocular pressure in rabbits.

In vivo central actions of NPY(1-30), an N-terminal fragment of neuropeptide Y.

The purpose of the present study was to examine the possible central actions of a N-terminal fragment of NPY, NPY(1-30), on five measures typically influenced by the native peptide: decreased spontaneous activity, enhancement of muscle tone (increased grasping response), catalepsy, hypothermia, and stimulation of food intake. The peptides were administered ICV in doses ranging from 2.5 to 160 micrograms (0.75-48 nmol) and their effects on the three motor variables as well as thermal and feeding responses were evaluated and compared. Globally, results indicate that, similarly to NPY, the N-terminal fragment NPY(1-30), decreased spontaneous activity and induced hypothermia. However, the fragment displayed approximately half of the potency of NPY for producing these effects. On the other hand, contrary to NPY, NPY(1-30) did not affect muscle tone or food consumption and did not induced catalepsy in animals. These results demonstrate for the first time central actions of a N-terminal fragment of NPY and lend further support to the hypothesis that the receptors mediating the central actions of NPY are pharmacologically different.

Effects of experimentally induced ischemia on dopamine metabolism in rabbit retina.

PURPOSE: To determine the role of dopamine in experimentally induced retinal ischemia. Experiment 1 was designed to study the effects of ischemic levels on dopamine (DA) metabolism. Experiment 2 evaluated the effects of ischemic duration on DA metabolism. The effects of recirculation time after ischemia on DA metabolism were investigated in experiment 3. METHODS: Ischemia was produced by raising intraocular pressure (IOP) in rabbits. Three levels of ischemia were used--level A, level B, and level C--representing 50%, 75%, and 100%, respectively, of the IOP necessary to produce total ischemia. Retinal levels of DA and its main metabolites, homovanillic acid (HVA) and 3,4-dihydroxyphenylacetic acid (DOPAC), were measured by high-performance liquid chromatography with electrochemical detection. RESULTS: Experiment 1 showed that at ischemic level B, DA contents were significantly reduced, but neither DOPAC nor HVA concentrations were altered. The reductions of retinal DA, DOPAC, and HVA concentrations were seen at level C without altering the ratio index. Level A ischemia did not alter DA metabolism. In experiment 2, significant reductions in DA, DOPAC, and HVA concentrations were found at both 30 and 60 minutes in the ischemic group, whereas the ratio DOPAC/DA and (HVA + DOPAC)/DA were significantly decreased only in the 60-minute ischemic group. Five-minute total ischemia did not alter DA metabolism. In experiment 3, concentrations of DA were still significantly decreased at 30 minutes of recirculation after ischemia, but DOPAC and HVA levels were back to normal. However, the ratios of DOPAC/DA, HVA/DA, and (HVA + DOPAC)/DA were significantly elevated. After 90 and 240 minutes of recirculation, retinal DA contents had returned to normal values, but DOPAC and HVA concentrations as well as all ratio indices of DA metabolism were still enhanced. CONCLUSION: Dopamine metabolism is altered during and after retinal ischemia. Dopamine may play a contributing role in ischemia.

Effects of calcitonin gene-related peptide on the rabbit electroretinogram.

In order to better understand the role of Calcitonin gene-related peptide (CGRP) in mammalian retina, the dose related effects of human CGRP (hCGRP) on rabbit electroretinogram (ERG) were examined in the present study. CGRP was administered intraocularly in doses of 0.1, 1.0 and 10.0 micrograms. ERG A- and B-wave as well as oscillatory potentials (P1, P2, P3 and P4) were recorded. The highest dose of CGRP (10.0 micrograms) significantly increased the amplitudes of the A-wave and OP components (P1, P2, P3 and P4) produced by relatively high stimulus intensity. The same dose of the peptide also enhanced B-wave amplitude at all intensities studied. The effects of the intermediate dose of CGRP (1.0 microgram) on the B-wave amplitudes were dependent on stimulus intensities. B-wave amplitudes at high stimulus intensities were not affected by 1.0 microgram of CGRP but were significantly increased with relatively lower stimulus intensities. The amplitudes of P3, one of OP components, were significantly increased. However, amplitudes of A-wave and other OP components (P1, P2 and P4) were not affected by 1.0 microgram CGRP. The lowest dose of the peptide (0.1 microgram) did not affect any amplitudes of ERG components. Implicit times of A-wave, B-wave and OP components were not significantly affected by the different doses of CGRP. Taken together, these results indicate that CGRP may play a functional role in modulating retinal responses to photic stimulation.

Dextromethorphan attenuates the effects of ischemia on rabbit electroretinographic oscillatory potentials.

Dextromethorphan has been shown to protect against ischemic tissue damage. We investigated the effects of dextromethorphan on electroretinographic oscillatory potentials in retinal ischemia. Retinal ischemia was induced in rabbits by increasing intraocular pressure to 120 mm Hg for 30, 60 or 90 minutes. Dextromethorphan was intravenously administered before ischemia and maintained throughout the whole period of experiments. Oscillatory potentials were recorded before and during ischemia as well as 4 hours of recirculation after ischemia. As expected, all oscillatory potentials were decreased after 60 and 90 minutes of ischemia. However, after 30 minutes of ischemia followed by 4 hours of recirculation, amplitudes of P2 were elevated whereas those of P3 and P4 were decreased with normal P1 amplitudes. Dextromethorphan administration diminished the effects of 30 minutes of ischemia on oscillatory potentials and partially attenuated the effects of 60 minutes of ischemia, whereas the effects of 90 minutes of ischemia could not be reversed by dextromethorphan treatment. These results indicate that electroretinographic oscillatory potentials could be useful indicators to evaluate retinal function in the ischemic condition and that dextromethorphan can attenuate the effects of relatively short periods of ischemia on rabbit electroretinographic oscillatory potentials.

Atypical neuroleptic-like behavioral effects of neurotensin.

To better characterize the neuroleptic-like properties of neurotensin, the dose-related effects of the peptide on the following behavioral phenomena were examined: a) the yawning-penile erection syndrome induced by small doses of the dopamine agonists apomorphine and N-propylnorapomorphine (NPA); b) yawning produced by the anticholinesterase physostigmine, and c) stereotyped climbing and sniffing produced by a larger dose of apomorphine. Several doses of the peptide were injected intraventricularly 30 min prior to drug administration. Results indicate that neurotensin markedly decreased yawning and penile erections produced by both apomorphine and NPA. These effects were seen with relatively small doses (0.9-3.75 micrograms). Neurotensin also potently decreased physostigmine-induced yawning with the initial inhibitory effect seen with 50 ng of the peptide. Apomorphine-induced climbing was significantly attenuated with 30.0 and 60.0 micrograms neurotensin, whereas stereotyped sniffing was unaffected, even by doses as large as 120.0 micrograms. These findings suggest that neurotensin might antagonize dopamine autoreceptors and indicate that the peptide possess central anticholinergic activity. Furthermore, these results lend support to the hypothesis that neurotensin's profile of central actions resemble that of atypical neuroleptics.