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The gradual recovery of activity of serotonergic neurons following an initial inhibition has been hypothesized to play an important role in the delayed onset of efficacy of selective serotonin reuptake inhibitors. This study explored the clinical relevance of this hypothesis by examining the effects of different doses and routes of administration of fluoxetine on the recovery of activity of serotonergic neurons over the course of a 21-day exposure. Single-unit, extracellular recordings of serotonergic neurons were made in the dorsal raphe nucleus of anesthetized male rats. Acute i.v., s.c. and i.p. administration of fluoxetine inhibited the activity of serotonergic neurons. With chronic administration of fluoxetine, at clinically relevant doses, the activity of serotonergic neurons gradually recovered to baseline levels over the course of 14-21 days. The dose of fluoxetine (5, 10 or 20 mg/kg per day) did not make a significant difference in the time course of the recovery of activity of serotonergic neurons. A significant, non-parallel shift in the dose-response curve of serotonergic neurons to the serotonin-1A (5-HT1A) agonist 8-OH-DPAT occurred over the 21 days of treatment with fluoxetine, indicating a desensitization of the 5-HT1A receptor during this period. The recovery of firing did not correlate with either plasma or cerebrospinal fluid levels of fluoxetine or norfluoxetine. These results indicate that, similar to the effects of dose on the speed of onset of the clinical effects of SSRIs, increasing the dose of fluoxetine does not hasten the recovery of firing of serotonergic neurons during chronic administration. These results support the hypothesis that desensitization of the 5-HT1A receptor and consequent recovery of firing of 5-HT cells in the dorsal raphe nucleus plays a role in the delayed therapeutic onset of fluoxetine.

LY426965 [(2S)-(+)-1-cyclohexyl-4-[4-(2-methoxyphenyl)-1-piperazinyl]2-methyl- 2-phenyl-1-butanone monohydrochloride] is a novel compound with high affinity for the cloned human 5-hydroxytryptamine (HT)(1A) receptor (K(i) = 4.66 nM) and 20-fold or greater selectivity over other serotonin and nonserotonin receptor subtypes. Both in vitro and in vivo studies indicate that LY426965 is a full antagonist and has no partial agonist properties. LY426965 did not stimulate [(35)S]guanosine-5'-O-(3-thio) triphosphate (GTPgammaS) binding to homogenates of cells expressing the cloned human 5-HT(1A) receptor in vitro but did inhibit 300 nM 5-HT-stimulated [(35)S]GTPgammaS binding with a K(i) value of 3.07 nM. After both p.o. and s.c. administration, LY426965 blocked the lower lip retraction, flat body posture, hypothermia, and increase in rat serum corticosterone induced by the 5-HT(1A) agonist 8-OH-DPAT (8-hydroxy-2-dipropylaminotetralin). In pigeons, LY426965 dose-dependently blocked the stimulus cue induced by 8-OH-DPAT but had no 8-OH-DPAT-like discriminative properties. LY426965 completely reversed the effects of nicotine withdrawal on the auditory startle reflex in rats. In microdialysis experiments, LY426965 administered together with fluoxetine significantly increased extracellular levels of serotonin above those achievable with fluoxetine alone. In electrophysiological studies, the administration of LY426965 produced a slight elevation of the firing rate of 5-HT neurons in the dorsal raphe nucleus of anesthetized rats and both blocked and reversed the effects of fluoxetine on 5-HT neuronal activity. These preclinical results indicate that LY426965 is a selective, full 5-HT(1A) antagonist that may have clinical use as pharmacotherapy for smoking cessation and depression and related disorders.

In order to explore the neurophysiology of nicotine withdrawal, we examined the activity of serotonergic neurons in the dorsal raphe nucleus in rats undergoing withdrawal from chronic exposure to nicotine. Animals were exposed to nicotine (6 mg/kg per day base) via SC implanted osmotic minipumps. After 12 days the pumps were removed and the animals allowed to go through spontaneous withdrawal. Rats were anesthetized on various days of the procedure and the effect of the 5-hydroxytryptamine (5-HT)-1A agonist 8-OH-DPAT on the single-unit activity of serotonergic neurons in the dorsal raphe nucleus was examined. The sensitivity of serotonergic neurons to 8-OH-DPAT was not changed by the chronic administration of nicotine or saline; slightly increased on day 2 of withdrawal; significantly increased on days 3 and 4 of withdrawal; and no longer significantly increased by day 7 of withdrawal. These results indicate that serotonergic neurons in the dorsal raphe nucleus have an increased sensitivity to systemically administered 8-OH-DPAT in rats undergoing nicotine withdrawal and that the serotonergic system may play a role in the symptoms of nicotine withdrawal.

Pre-pulse inhibition (PPI) of the acoustic startle response is the diminution of the startle response when the startle stimulus is preceded by a weaker, non-startle-eliciting stimulus. Deficits in PPI occur in animals following the administration of apomorphine and in schizophrenic patients. In this study, we examined the ability of the novel atypical antipsychotic olanzapine and the cholecystokinin (CCK)-B antagonist LY288513 to reverse the apomorphine-induced disruption of pre-pulse inhibition. Olanzapine (3.0 and 5.0 mg/kg, i.p.), but not LY288513 (1.0-100 mg/kg, p.o.), blocked apomorphine-induced disruption of PPI. These results indicate that olanzapine, but not LY288513, has dopamine antagonist properties in vivo and predict that olanzapine, but not LY288513, will have antipsychotic activity in man.

In order to explore the potential clinical utility of CCK-B antagonists for the treatment of nicotine withdrawal symptoms, the auditory startle reflex was examined in rats undergoing withdrawal from the chronic administration of nicotine. Rats were exposed to nicotine continuously for 12 days (6 mg kg-1 day-1) via osmotic minipumps. After 12 days the pumps were removed and the animals allowed to go through spontaneous withdrawal for several days. Acute treatment with the CCK-B antagonist LY288513, at doses that have no effect on startle responses in naive rats, blocked the nicotine withdrawal-induced increase in the acoustic startle reflex. These results indicate that CCK-B antagonists may be an efficacious treatment for some nicotine withdrawal symptoms in man and may represent a novel pharmacotherapy for smoking cessation.

The activity of LY288513, an investigational cholecystokinin (CCK)B antagonist, was evaluated in a wide range of pharmacological tests in mice and rats. The anxiolytic benzodiazepine, diazepam, served as a reference standard for LY288513 in many of the tests. In the elevated plus-maze, LY288513 (3, 10 mg/kg, IP; 10, 30 mg/kg, PO) produced an anxiolytic-like action in mice with a magnitude of effect similar to that of diazepam. However, unlike diazepam, LY288513 produced no overt clinical signs and did not affect muscle tone, neuromuscular coordination, or sensorimotor reactivity. Also, in contrast to diazepam, LY288513 did not produce changes in the thresholds for electroshock- or pentylenetetrazol-induced convulsions. High doses of LY288513 (1000 mg/kg, PO) were required to reduce spontaneous activity levels, decrease body temperature, or potentiate the CNS-depressant effects of hexobarbital. LY288513 had no analgesic activity in mouse writhing or tail-flick tests. Electrophysiological studies in anesthetized rats showed that acute administration of LY288513 decreased the number of spontaneously active dopamine neurons in the substantia nigra and ventral tegmental area. However, LY288513 did not produce catalepsy. These data indicate that LY288513 possess both anxiolytic and antipsychotic potential.

In order to explore the neurophysiology of nicotine withdrawal, we examined the activity of substantia nigra (A9) and ventral tegmental area (A10) dopamine cells in rats undergoing withdrawal from chronic exposure to nicotine. Animals were exposed to nicotine (6 mg kg-1 day-1 base) via s.c. implanted osmotic minipumps. After 12 days the pumps were removed and the animals allowed to go through spontaneous withdrawal. Rats were anesthetized on various days of the procedure and single-unit recordings were made from A9 and A10 dopamine cells. Chronic administration of nicotine led to a decreased firing rate of A10, but not A9, dopamine cells. Upon withdrawal from the chronic exposure to nicotine, the firing rates of A10 dopamine cells returned to control levels, while the firing rate of A9 dopamine cells significantly increased above control levels. This increased dopamine neuronal activity may play a role in some behavioural symptoms of nicotine withdrawal.

The diphenylpyrazolidinone cholecystokinin (CCK)-B antagonist LY262691 has recently been demonstrated to decrease the number of spontaneously active dopamine (DA) cells in the ventral tegmental area (A10) and substantia nigra (A9) of the anesthetized rat. In the present study, three structural analogs of LY262691 with high selectivity for CCK-B receptors, LY262684, LY191009 and LY242040, also decreased the number of spontaneously active A10 DA cells. Neither an inactive analog (LY206890) nor a CCK-A-selective analog (LY219057) affected the number of spontaneously active A10 DA cells. L-365,260, a benzodiazepine CCK-B antagonist, also decreased the number of spontaneously active A10 DA cells. In addition, the more active optical isomer of LY262691 (LY288513) caused twice as large a decrease in the number of spontaneously active A10 DA cells as the less active optical isomer (LY288512). The diphenylpyrazolidinone CCK-B antagonists, but neither the inactive nor the CCK-A selective analog, also decreased the number of spontaneously active A9 DA cells; however, none of these compounds produced catalepsy in awake animals. Single-unit recordings indicated that LY262691 administration inhibited the activity of individual A9 and A10 DA neurons. These results indicate that the firing of A9 and A10 DA neurons is suppressed specifically by antagonism of CCK-B, but not CCK-A receptors. CCK-B antagonists may therefore represent a novel class of antipsychotic drugs. Furthermore, because CCK-B antagonists have no cataleptogenic effects, they may also have a reduced propensity for producing extrapyramidal side effects. In addition, these actions on midbrain DA neurons may contribute to the known anxiolytic activity of CCK-B antagonists.

Chronic administration of antipsychotic drugs has previously been shown to decrease the number of spontaneously active midbrain dopamine cells. In an effort to evaluate CCK antagonists as potential antipsychotic drugs, we have examined the effects of a selective CCK-B antagonist, LY262691, on the number of spontaneously active midbrain dopamine neurons using extracellular, single-unit recordings in anesthetized rats. Acute and chronic administration of LY262691 decreased the number of spontaneously active A9 and A10 dopamine cells. Administration of apomorphine did not reverse the effect of LY262691 on A9 and A10 dopamine neurons. These results suggest that LY262691 may have an antipsychotic effect without delayed onset, and that its effects on dopamine cells may not be mediated through depolarization inactivation.

Acute and chronic administration of low doses (e.g. 0.1, 0.3 mg/kg i.p.) of the selective 5-HT3 antagonist zatosetron decreased the number of spontaneously active A 10 dopamine cells but did not change the number of spontaneously active A9 dopamine cells; higher doses (1.0, 10 mg/kg) were less effective. The decrease in the number of spontaneously active A 10 dopamine cells following zatosetron administration was not reversed by the administration of apomorphine. These data indicate that zatosetron's effects on spontaneously active dopamine neurons: (1) differs from other 5-HT3 antagonists; (2) may not be mediated by depolarization inactivation; and, (3) may be predictive of an atypical antipsychotic action without delayed onset.

Acute administration of ethanol is accompanied by alterations in dopamine turnover and release, and chronic ethanol exposure is associated with changes in biochemical measures of dopamine receptor function. This paper presents data examining the effects of chronic ethanol exposure on behavioral responses to the dopamine receptor agonist apomorphine. Measurements of behavior were obtained through the use of an electronic motility monitor which permitted the quantification of movements in terms of their characteristic frequency components. Results are presented which indicate that apomorphine-induced movements with modal frequencies of 2 Hz and of 8-9 Hz are significantly increased during the 12 to 24 hr following ethanol withdrawal, suggesting an increase in the functional responsiveness of central dopaminergic systems.

Changes in several measures of dopamine function have been observed following acute or chronic ethanol exposure. The present study examined the effects of chronic ethanol exposure on the hypothermia following acute administration of the dopamine agonist apomorphine. Animals withdrawn from chronic ethanol exposure showed a significantly greater decrease in body temperature following apomorphine than did ethanol-naive controls, suggesting an increase in sensitivity to dopaminergic stimulation during ethanol withdrawal.

Increased biochemical measures of GABA activity are observed after acute administration of ethanol and decreased activity has sometimes been observed after chronic ethanol exposure. Since chronic alterations in neurotransmitter activity may result in changes in receptor function, it is possible that changes in GABA-receptive neurons may accompany chronic ethanol treatment. In the present study we examined the incidence of muscimol-induced motor behaviors in ethanol-naive and chronic ethanol-treated animals. Male Sprague-Dawley rats received bilateral cannula implants into substantia nigra pars reticulata for subsequent administration of muscimol or saline. After recovery from surgery, rats received chronic treatment in ethanol-vapor inhalation chambers for 15 days. Animals were then removed from the chambers and examined 10 hours after removal. Muscimol resulted in a general increase in motility in both control and ethanol-treated animals. Animals withdrawn from chronic ethanol exposure, however, exhibited significantly less muscimol-stimulated, repetitive 9 Hz movements. These results suggest that GABA receptive cells within the substantia nigra or its vicinity may be functionally less responsive to GABAergic stimulation after chronic ethanol administration.

Ethanol withdrawal symptoms in clinical populations are observed to occur in unique clusters which differ in time of onset relative to the time of withdrawal and in their duration. Since periods of mild symptoms are sometimes observed between these clusters of more severe symptoms, the symptom clusters may reflect separate periods of hyperexcitability during which times different neuronal mechanisms are involved. To investigate this possibility in an animal model of ethanol withdrawal, rats were chronically exposed to ethanol in vapor inhalation chambers. Upon withdrawal from this exposure regimen, the time course of spontaneous seizure activity was observed for a period of 84 hr and compared to sensitivity to seizures elicited by audiogenic stimuli or by the convulsant drug picrotoxin. Spontaneous seizure events were observed to occur in clusters, and these clusters were differentially correlated with periods of increased sensitivity to induced seizure activity. These results further support the suggestion that seizure sensitivity during ethanol withdrawal may indicate the involvement of multiple, independent, neuronal mechanisms.