Nicotine In Brain Research Articles

Administration of nicotine to adolescent rats evokes regionally selective upregulation of CNS α7 nicotinic acetylcholine receptors

α7 Nicotinic acetylcholine receptors (nAChRs) play a role in axonogenesis, synaptogenesis and synaptic plasticity, and are therefore targets for developmental neurotoxicants. We administered nicotine to adolescent rats and evaluated the effects on α7 nAChRs in the striatum, brainstem and cerebellum. During the period of nicotine administration (30–47.5 days of age), nicotine elicited α7 nAChR upregulation with a regional hierarchy of striatum>brainstem>cerebellum. Values returned to normal or became slightly subnormal almost immediately after the cessation of treatment (50 days of age) with no further changes through 75 days of age. The temporal and regional patterns of the effects on α7 nAChRs were distinct from those reported earlier for the α4β2 subtype, and neither adult nor fetal/neonatal administration upregulates the α7 subtype in the striatum. Targeting of the striatum is thus unique to nicotine exposure during adolescence and parallels earlier work showing regionally selective effects of this treatment on synaptic signaling. We obtained preliminary evidence for nicotine-induced oxidative stress as a potential contributory mechanism. The present findings reinforce the concept of biologically distinct effects of nicotine in the adolescent brain and provide evidence for a mechanistic involvement of α7 nAChRs in its unique effects during this developmental period.

Dementia rating and nicotinic receptor expression in the prefrontal cortex in schizophrenia

Background The etiology of dementia that occurs in patients with schizophrenia is not well understood. Nicotinic acetylcholine receptors have been implicated in cognitive function, and deficits in these receptors have been reported in schizophrenia. Methods The present study investigates possible associations of nicotinic receptor subunit expression in the dorsal lateral prefrontal cortex, an area known to be affected in schizophrenia, and dementia rating. Results α7 immunoreactivity was reduced by 20% to 28% and [ 3H]epibatidine binding was increased twofold in groups of patients with schizophrenia compared to normal control subjects matched for age, postmortem delay, and low levels of brain nicotine and cotinine. In contrast, no significant differences in α4, α3, or β2 immunoreactivity or α7 messenger RNA expression were observed in schizophrenia patients compared with control subject values. Clinical dementia ratings in patients with schizophrenia were correlated with neither [ 3H]epibatidine binding nor nicotinic receptor subunit expression. Conclusions These data indicate no relationship between the trend for reduced neocortical α7 subunit protein expression in schizophrenia and dementia. Further investigations are required to establish whether the reduction in α7 protein in the dorsal lateral prefrontal cortex is associated with clinical features other than dementia in schizophrenia.

Nicotine accumulation in the mouse brain is age-dependent and is quantitatively different in various segments

Mice, varying from newborn to adult mice, were injected intraperitoneally with 1 mg/kg of 14C-nicotine. The nicotine concentration in blood and different segments of the brain (Cerebellum, Pons and Medulla oblongata, Midbrain, B. olfactory, Hypothalamus, Hippocampus and Cortex) was determined in newborn (0-day old); in 1-, 3-, 5-, 7-, 9-, 12- and 17-day old; and in adult (26-day old) mice. The nicotine concentration in the blood did not differ in mice aged 0–12 days, but the concentration thereafter decreased substantially until adult age. The decrease was associated with metabolism of nicotine as evidenced by cotinine formation, as well accumulation in different regions of the brain. Irrespective of the small continuous brain growth from day 1 until adult age, the age dependent nicotine uptake was dramatically increased in all segments of the brain between 9 and 12 days of age. However, the various brain regions showed differences in magnitude of growth and age-dependent changes in the distribution of nicotine. The relative nicotine concentration in Pons Med. obl. seems to be high in young mice until 9 days of age; thereafter, the concentration decreases in comparison to the other regions. Conversely, the relative concentration in the hippocampus and cortex rises from day 9 to 17 of age. The present results show that the ability of the brain to accumulate nicotine increases with age, reaching a peek at day 12 of age. Furthermore, this study for the first time shows that the age-dependent brain nicotine accumulation is quantitatively different in the various segments of the brain.

Tissue-dependent effects of immunization with a nicotine conjugate vaccine on the distribution of nicotine in rats

Vaccination of rats against nicotine reduces nicotine distribution to brain even at nicotine doses greatly exceeding the estimated binding capacity of the available antibody. This observation suggests a differential effect by which vaccination reduces nicotine distribution to brain to a greater extent than to other tissues. To test this hypothesis, vaccinated rats received a single intravenous nicotine dose equal to twice the estimated binding capacity of nicotine-specific antibody in vaccinated rats. The total and bound serum nicotine concentrations were higher in the vaccinated rats compared to controls, while the unbound serum nicotine concentration was lower. Distribution of nicotine to brain was reduced in vaccinated rats in a time-dependent manner, with a greater reduction at 1 min (64%) than at 25 min (45%). Vaccination reduced nicotine distribution to muscle, testis, spleen, liver, heart, and kidney, but to a lesser extent than to brain, while nicotine distribution to fat was increased. Chronically infused nicotine showed a similarly altered pattern of tissue distribution in vaccinated rats, but differences were in general smaller than after a single nicotine dose; brain nicotine concentration was 24% lower in vaccinated rats, while lung nicotine concentration was higher. The presence of nicotine-specific antibody in tissues may have contributed to the increased nicotine concentrations in fat and lung. These data suggest that vaccination reduces nicotine distribution to brain not only by sequestering nicotine in serum but also by redirecting tissue distribution disproportionately away from brain, such that nicotine concentrations are reduced to a greater extent in brain than in other tissues.

Maternal vaccination against nicotine reduces nicotine distribution to fetal brain in rats.

Cigarette smoking during pregnancy is associated with a variety of adverse fetal outcomes. Nicotine is a likely contributor to these adverse effects, with fetal brain as one target organ. Vaccination of adult male rats against nicotine has been shown to reduce nicotine distribution to the brain. The current study examined whether vaccination of female rats before pregnancy would reduce the distribution to fetal brain of a single nicotine dose administered during gestation. Female rats immunized with a nicotine conjugate vaccine received a single dose of nicotine 0.03 mg/kg i.v. on gestational day 16 to 22. Five minutes later, vaccinated rats had substantially higher bound and lower unbound serum nicotine concentration and lower brain nicotine concentration than controls. Fetal brain nicotine concentration was reduced by 43% in vaccinated rats, comparable to the reduction in the maternal brain nicotine concentration. The whole-fetus nicotine concentration was not altered by vaccination. A similar experiment was performed in which pregnant rats were passively immunized with rabbit nicotine-specific IgG 7 or 21 mg/kg just before nicotine dosing. The effects of passive immunization on nicotine distribution in the mother were IgG dose-related and the higher dose reduced nicotine distribution to fetal brain by 60%. These data suggest that vaccine effects on nicotine distribution to serum and brain are similar in pregnant female rats to those previously reported in adult males. Vaccination of female rats before pregnancy, or passive immunization during pregnancy, can reduce the exposure of fetal brain to a single dose of maternally administered nicotine.

Detrimental Effects of Nicotine and Endotoxin in the Newborn Piglet Brain during Severe Hypoxemia

Hypoxia-ischemia is a major cause of perinatal brain damage, but evidence shows that brain injury also is associated with intrauterine infections and maternal smoking. The mechanisms are not known, and we therefore explored the effects of experimental inflammation or nicotine on perinatal brain metabolism and injury during severe hypoxemia. Twenty-eight 1-week-old piglets were anesthetized and instrumented with microdialysis probes in the striatum and brainstem. We studied three pretreatment groups: (1) 20 µg/kg i.v. nicotine (n = 9); (2) 1 µg/kg i.v. endotoxin from Escherichia coli (n = 11), or (3) control (n = 8). The piglets were subsequently exposed to 30 min of hypoxemia (6% O₂). In order to minimize any ischemic component and increase survival, this was abrupted for 1 min if blood pressure fell to 30 mm Hg. During hypoxemia, both the pretreatment with endotoxin and nicotine induced higher levels of extracellular lactate and peak lactate/pyruvate ratio compared with controls (54.7 ± 9.6 (p < 0.01) and 65.2 ± 13.1 (p < 0.02) vs. 15.9 ± 7.4, respectively), reflecting a deterioration of the metabolic status in these groups. The two pretreated groups reached significantly higher peak levels of extracellular glycerol (30.9 ± 4.1 vs. 77.9 ± 12.7 and 89.4 ± 14.2 µmol/l, respectively, p = 0.01), indicating a higher level of cellular membrane disintegration or leakage. In addition, 3 endotoxin piglets and 4 nicotine piglets died during reoxygenation, while all controls survived (p = 0.13 and p < 0.04, respectively). Mortality was associated with a rise in extracellular glutamate at the end of hypoxemia/start reoxygenation (p = 0.02). These findings contribute in explaining how nicotine and inflammatory response to bacterial toxins could act as cofactors for hypoxic-ischemic neurologic injury in the immature brain.

Active Immunization against Nicotine Prevents Reinstatement of Nicotine-Seeking Behavior in Rats

Background: The presently available pharmaceutical aids in smoking cessation possess a rather limited effectiveness. Therefore, we have synthesized a series of immunoconjugates that stimulate the induction of antibodies which may bind nicotine in the blood, thereby preventing it from passing the blood-brain barrier. Thus, the reinforcing action of nicotine in the brain, which is the driving force in tobacco smoking, should be abolished. Objective: The present study was undertaken to test this notion in a long-term relapse model in rats, measuring the reinstatement of nicotine-seeking behavior, following active immunization with IP18-KLH, one of our immunoconjugates. Methods: Male Wistar rats were immunized with a nicotine-KLH conjugate (nicotine immunogen) and Freund’s adjuvant after having been trained to meet the criteria of stable nicotine self-administration on a fixed ratio (FR3) schedule. The rats were subsequently extinguished from nicotine self-administration behavior and finally, as extinction was completed, they were exposed to small, priming doses of nicotine, which previously have been shown to reinstate the nicotine-seeking behavior. The antibody titers were measured by ELISA. Results: It was found that rats with high titers (>1:10,000) of antibodies against nicotine, in contrast to those with low/no nicotine selective antibodies, do not reinstate nicotine self-administration behavior when they are exposed to nicotine. Conclusions: Our findings indicate that active immunization against nicotine may effectively abolish the reinforcing action of nicotine in brain, an effect which is critical for relapse in nicotine dependence. These data suggest the potential utility of active immunization in smoking cessation programs.

The Selectively Bred High Alcohol Sensitivity (HAS) and Low Alcohol Sensitivity (LAS) Rats Differ in Sensitivity to Nicotine

Studies in rodents selectively bred to differ in alcohol sensitivity have suggested that nicotine and ethanol sensitivities may cosegregate during selective breeding. This suggests that ethanol and nicotine sensitivities may in part be genetically correlated. Male and female high alcohol sensitivity (HAS), control alcohol sensitivity, and low alcohol sensitivity (LAS) rats were tested for nicotine-induced alterations in locomotor activity, body temperature, and seizure activity. Plasma and brain levels of nicotine and its primary metabolite, cotinine, were measured in these animals, as was the binding of [3H]cytisine, [3H]epibatidine, and [125I]alpha-bungarotoxin in eight brain regions. Both replicate HAS lines were more sensitive to nicotine-induced locomotor activity depression than the replicate LAS lines. No consistent HAS/LAS differences were seen on other measures of nicotine sensitivity; however, females were more susceptible to nicotine-induced seizures than males. No HAS/LAS differences in nicotine or cotinine levels were seen, nor were differences seen in the binding of nicotinic ligands. Females had higher levels of plasma cotinine and brain nicotine than males but had lower brain cotinine levels than males. Sensitivity to a specific action of nicotine cosegregates during selective breeding for differential sensitivity to a specific action of ethanol. The differential sensitivity of the HAS/LAS rats is due to differences in central nervous system sensitivity and not to pharmacokinetic differences. The differential central nervous system sensitivity cannot be explained by differences in the numbers of nicotinic receptors labeled in ligand-binding experiments. The apparent genetic correlation between ethanol and nicotine sensitivities suggests that common genes modulate, in part, the actions of both ethanol and nicotine and may explain the frequent coabuse of these agents.

Impact of intravenous nicotine on BOLD signal response to photic stimulation

Functional magnetic resonance imaging (fMRI) is increasingly being applied in the study of brain effects of nicotine. In addition, because tobacco smoking is common, many subjects studied with fMRI for other reasons may have appreciable levels of nicotine in plasma and brain during scanning. However, there is concern that the vascular effects of nicotine may alter the coupling between blood oxygen level dependent (BOLD) signal and neuronal activity. The objective of this study was to test for evidence of alteration of BOLD signal response of occipital cortex, a region with a relatively low concentration of neuronal nicotine receptors, to photic stimulation during intravenous infusion of nicotine. Nine nicotine dependent healthy smokers were withdrawn from nicotine under controlled conditions and then scanned while receiving photic stimulation and successive intravenous infusions of saline and nicotine. No evidence for an effect of nicotine on BOLD signal response to photic stimulation was detected at the doses studied. This observation suggests that nicotine does not alter the coupling between BOLD signal and neuronal activity in the visual cortex.

Solid-Phase Extraction and HPLC Assay of Nicotine and Cotinine in Plasma and Brain

The aim of this study was to develop a simple and reliable assay for nicotine (NIC) and its major metabolite, cotinine (COT), in plasma and brain. A method was developed that uses an extraction method compatible with reverse-phase high-performance liquid chromatography (HPLC) separation and ultraviolet (UV) detection. Sequential solid-phase extraction on silica columns followed by extraction using octadecyl (C18) columns resulted in mean percent recovery (n = 5) of 51 ± 5, 64 ± 10, and 52 ± 10% for NIC, COT, and phenylimidazole (PI), respectively, in spiked 1-mL serum samples. Recovery (mean ± SEM) of the internal standard (PI) from spiked samples of nicotine-injected rats averaged 64.1 ± 1.5% (n = 138) from plasma, and 20.7±0.8% (n = 128) from brain. The limits of detection of NIC in plasma samples were approximately 8 ng per mL, and of COT, 13.6 ng per mL. Further optimization of our extraction method, using slower flow rates and solid-phase extraction on silica columns, followed by C18 column extraction, yielded somewhat better recoveries (38 ±3%) for 1-mL brain homogenates. Interassay precision (coefficient of variation) was determined on the basis of daily calibrations for 2 months and was found to be 7%, 9%, and 9% for NIC, COT, and PI, respectively, whereas intra-assay variability was 3.9% for both NIC and COT. Limited studies were performed on analytical columns for comparison of retention, resolution, asymmetry, and column capacity. We concluded that a simple two-step solid-phase extraction method, coupled with HPLC separation and UV detection, can be used routinely to measure NIC and COT in biological fluids and tissues.

Enhanced motor activity and brain dopamine turnover in mice during long-term nicotine administration in the drinking water

Nicotine was administered chronically to NMRI mice in their drinking water in gradually increasing concentrations to measure gross motor activity and brain nicotine concentrations over 24 h on the 50th day of nicotine administration. Also, the striatal postmortem tissue concentrations and accumbal extracellular concentrations of dopamine (DA) and its metabolites 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) were measured to study the role of dopaminergic systems in nicotine-induced hyperactivity in mice. The cerebral nicotine concentration was at its highest at the end of the dark period. The activity of nicotine-treated mice and their striatal DA metabolism were parallelly increased at 2 to 3 h after midnight and in the forenoon. Microdialysis experiments carried out in the forenoon showed that the extracellular levels of DA and DOPAC were elevated in the nucleus accumbens of these mice. Nicotine did not alter the circadian rhythmicity of activity in the mice. Rather, our findings suggest that the mice consume more nicotine when active and this might lead to enhanced release and metabolism of DA and further, to enhanced motor behavior. These findings support the suggestions that nicotine's effects on limbic and striatal DA are critical for its stimulating effects.

Induction of Fos-immunostaining by nicotine and nicotinic receptor antagonists in rat brain

Using Fos protein immunohistochemistry, we have studied the effects of acute nicotine (0.5 mg/kg s.c.) and nicotinic acetylcholine receptor (nAChR) antagonists in eleven rat brain areas. Acute nicotine elevated Fos-like immunostaining (Fos IS) significantly in all studied areas except the medial prefrontal cortex. Nicotine increased the Fos IS in cortical, limbic and hypothalamic areas by 2–10-fold, and in the interpeduncular nucleus as well as in the visual areas the increases were 15–150-fold. When given alone, the nAChR antagonists mecamylamine (1.0 or 5.0 mg/kg i.p.) and dihydro-beta-erythroidine (DHE; 1.4 or 2.8 mg/kg i.p.) increased Fos IS in most brain areas maximally by 2–10-fold, but methyllycaconitine (MLA; 4.0 mg/kg i.p.) only in three areas and maximally by 4-fold. The efficacy of nAChR antagonists in blocking nicotine's effects on Fos IS varied noticeably with respect to region and antagonist, and the combined effect of nicotine+antagonist did not exceed that of either treatment alone. Mecamylamine and DHE significantly reduced nicotine-induced Fos IS in most of the studied areas, and MLA only in two areas. Thus, nAChRs seem to mediate the effects of nicotine on Fos IS, and the differences in the effects of the antagonists studied suggest that more than one subtype of nAChRs are involved. The present experiments also provide evidence that nAChR blockade itself may result in increased Fos protein expression in the brain. This could be due to blockade of presynaptic nAChRs modulating transmitter release or interruption of complex polysynaptic feedback pathways.

Region-specific transcriptional response to chronic nicotine in rat brain

Even though nicotine has been shown to modulate mRNA expression of a variety of genes, a comprehensive high-throughput study of the effects of nicotine on the tissue-specific gene expression profiles has been lacking in the literature. In this study, cDNA microarrays containing 1117 genes and ESTs were used to assess the transcriptional response to chronic nicotine treatment in rat, based on four brain regions, i.e. prefrontal cortex (PFC), nucleus accumbens (NAs), ventral tegmental area (VTA), and amygdala (AMYG). On the basis of a non-parametric resampling method, an index (called jackknifed reliability index, JRI) was proposed, and employed to determine the inherent measurement error across multiple arrays used in this study. Upon removal of the outliers, the mean correlation coefficient between duplicate measurements increased to 0.978±0.0035 from 0.941±0.045. Results from principal component analysis and pairwise correlations suggested that brain regions studied were highly similar in terms of their absolute expression levels, but exhibited divergent transcriptional responses to chronic nicotine administration. For example, PFC and NAs were significantly more similar to each other ( r=0.7; P<10 −14) than to either VTA or AMYG. Furthermore, we confirmed our microarray results for two representative genes, i.e. the weak inward rectifier K + channel (TWIK-1), and phosphate and tensin homolog (PTEN) by using real-time quantitative RT-PCR technique. Finally, a number of genes, involved in MAPK, phosphatidylinositol, and EGFR signaling pathways, were identified and proposed as possible targets in response to nicotine administration.

A Nicotine Conjugate Vaccine Reduces Nicotine Distribution to Brain and Attenuates Its Behavioral and Cardiovascular Effects in Rats

Vaccination of animals to elicit drug-specific antibodies, or the passive transfer of such antibodies from other animals, can reduce the behavioral effects of drugs such as cocaine and heroin. To study the potential application of this approach to treating nicotine dependence, IgG was isolated from rabbits immunized with a nicotine–protein conjugate vaccine. Anesthetized rats received immune IgG containing nicotine-specific antibodies (Nic-IgG) or control-IgG IV. Thirty minutes later, rats received nicotine at 0.03 mg/kg IV, equivalent on an mg/kg basis to the nicotine intake from two cigarettes by a smoker. Compared to control-IgG, Nic-IgG reduced the brain nicotine concentration in a dose-related manner (65% reduction at the highest IgG dose). Pretreatment with Nic-IgG also reduced the distribution to brain of five repeated doses of nicotine (equivalent to the nicotine intake from 10 cigarettes) administered over 80 min. To study blood pressure effects, rats received control-IgG or Nic-IgG 1 day prior to administering nicotine. Nicotine-induced systolic blood pressure increases were attenuated by Nic-IgG in a dose-related manner, and were almost completely blocked by the highest Nic-IgG dose. Pretreatment with Nic-IgG also completely prevented the nicotine-induced stimulation of locomotor activity observed in rats receiving control-IgG. Nic-IgG did not prevent locomotor activation from cocaine, demonstrating its specificity for nicotine. These data demonstrate that the administration of nicotine-specific antibodies can reduce or prevent some of the pharmacokinetic, cardiovascular, and behavioral consequences of nicotine in rats. Effects were observed at nicotine doses and nicotine serum concentrations equal to or exceeding those typically associated with nicotine exposure in cigarette smokers. A potential role for immunization in the treatment of nicotine dependence is suggested.

Altered disposition of repeated nicotine doses in rats immunized against nicotine.

Active immunization of rats against nicotine reduces the distribution of a single intravenous dose of nicotine to brain. The current study examined the effects of immunization on the distribution of repeated doses of nicotine, and on estimates of nicotine pharmacokinetic parameters. Groups of rats received five repeated doses of nicotine over 80 min (total dose equivalent to the nicotine intake from one, three or 10 cigarettes in a human). The serum nicotine concentration in immunized rats was 160-430% higher than controls after the fifth dose, demonstrating binding of nicotine to antibody. Brain nicotine concentration in immunized rats was reduced by 30-46%. The reduction in distribution of nicotine to brain correlated with the serum hapten-specific antibody concentration, the percentage of nicotine bound in serum, and with the unbound nicotine concentration in serum. In immunized rats, nicotine had a smaller steady state volume of distribution, lower systemic clearance, and longer terminal half-life than in controls. These data demonstrate that immunization against nicotine reduces nicotine distribution to brain, even after multiple nicotine doses at rates approximating heavy cigarette smoking. Whether this reduction in nicotine distribution is large enough to alter nicotine's physiological or behavioral effects remains to be studied.

Desnitroimidacloprid and Nicotine Binding Site in Rat Recombinant α4β2 Neuronal Nicotinic Acetylcholine Receptor

Desnitroimidacloprid (desnitro-IMI) is proposed to be a bioactivation product of imidacloprid and to bind at the same site as [3H]nicotine in the nicotinic acetylcholine receptor (nAChR) of mouse brain membranes. The α4β2 nAChR subtype accounts for >90% of the binding sites for nicotine in rat brain. This study further characterizes the binding site for [3]desnitro-IMI and [3H]nicotine in rat recombinant α4β2 nAChR using receptor expressed in Sf9 insect cells so that the assays involved no other receptor subtypes or interference from metabolic activation and detoxification systems. The two radioligands gave the same Bmax of 7.5 pmol/mg protein and apparent Kd values of 3.3 nM for nicotine and 8.9 nM for desnitro-IMI by Scatchard analysis at 22°C. However, at 4°C, the observed apparent association rate is slower and dissociation rate is faster for [3H]desnitro-IMI than for [3H]nicotine and due to the rapid rate of dissociation of [3H]desnitro-IMI the Kd calculated from the determined association and dissociation rates more closely approximates 1.0 for both ligands. Eight cholinergic agents and nine nicotinoids are equipotent in displacing [3H]desnitro-IMI and [3H]nicotine, with IC50 values (nM) of 0.5 for epibatidine, 1 for cytisine, 4–6 for nicotine and desnitro-IMI, 15 for acetylcholine, and 155 for imidacloprid, with an overall correlation for inhibitor potencies of r2 = 0.99 (n = 17). This correlation of binding site properties extends to [3H]nicotine in the recombinant α4β2 receptor and rat brain membranes (r2 = 0.99, n = 12). Thus, desnitro-IMI and nicotine bind with high affinity to the same site in rat recombinant α4β2 neuronal nAChR. This recombinant receptor can be generated in sufficient quantities for high-throughput target site screening and structural analysis of the binding site.

Immunization of rats reduces nicotine distribution to brain.

The effect of active immunization against nicotine on the initial distribution of nicotine to brain was studied in anesthetized rats. Animals received nicotine 0.03 mg/kg nicotine (equivalent to the nicotine dose absorbed by a human smoking two cigarettes) as a rapid injection in the jugular vein. In control animals, the arterial serum nicotine concentration initially exceeded the venous concentration 4.6-fold, similar to the initial arteriovenous difference produced by cigarette smoking in humans. Animals immunized with the nicotine analog CMUNic maintained this arteriovenous gradient, but with both arterial and venous nicotine concentrations several times higher than in controls. The arterial nicotine concentration was higher in immunized animals even at the first (7.5 s) sampling time. The brain nicotine concentration at 3 min was 36% lower in the immunized animals. The time course of nicotine distribution to brain was studied in a second group of animals. Brain nicotine concentration was reduced in rats immunized with CMUNic over the entire 6-min sampling period immediately following nicotine dosing (mean reduction 38%). A reduction was found at the earliest sampling time (30 s) and was maximal at 1 min (48%). Nicotine protein binding in serum was markedly increased in animals immunized with CMUNic compared to controls (91.2 versus 10.9%), and the unbound nicotine concentration in serum was lower (10.0 versus 13.4 ng/ml). The reduction in brain nicotine concentration correlated with antibody affinity for nicotine, and the percentage of nicotine bound in serum. These data demonstrate that nicotine-specific antibodies produced by active immunization rapidly bind nicotine in arterial blood, reduce the unbound nicotine concentration, and reduce the early distribution of nicotine to brain. Effects were observed using a clinically relevant nicotine dose and route of administration. These data suggest that the use of immunization to modify the behavioral effects of nicotine may be possible.

Imaging the neurochemistry of nicotine actions: studies with positron emission tomography.

Although the effects of nicotine in the brains of laboratory animals have been investigated extensively, very little is known about its effects in the human brain. With positron emission tomography (PET), a non-invasive imaging technology that allows measurement of the concentration of positron-labeled compounds that are of physiological and pharmacological relevance, it has become possible to investigate the effects of nicotine in the human brain. These imaging studies have shown that nicotine has very fast pharmacokinetics in the human brain, that it changes cerebral blood flow (CBF) and brain metabolism, and that at least some of these effects show acute tolerance. PET studies have also shown that, in addition to nicotine, cigarettes possess other pharmacological actions that may contribute to their reinforcing effects, that cigarettes inhibit monoamine oxidase (MAO) A and B in the brain, and that this inhibition recovers with cigarette discontinuation. Although the nicotine receptors have not yet been imaged in the living human brain, PET studies in the primate brain have shown very high concentration of receptors in the thalamus and a high rate of blockade by doses of nicotine that approximate plasma levels achieved by humans when smoking cigarettes. However, further studies are required to determine the levels of nicotine receptor occupancies achieved when smoking a cigarette and those required for the nicotine patch to be therapeutically effective, to measure the half-life for MAO inhibition by cigarettes and the mechanisms underlying this inhibition, and to evaluate the effects of smoking on nicotine receptors and on other neurotransmitter systems in the human brain.

Nicotine-induced limbic cortical activation in the human brain: a functional MRI study.

Nicotine is a highly addictive substance, and cigarette smoking is a major cause of premature death among humans. Little is known about the neuropharmacology and sites of action of nicotine in the human brain. Such knowledge might help in the development of new behavioral and pharmacological therapies to aid in treating nicotine dependence and to improve smoking cessation success rates. Functional magnetic resonance imaging, a real-time imaging technique, was used to determine the acute CNS effects of intravenous nicotine in 16 active cigarette smokers. An injection of saline followed by injections of three doses of nicotine (0.75, 1.50, and 2.25 mg/70 kg of weight) were each administered intravenously over 1-minute periods in an ascending, cumulative-dosing paradigm while whole brain gradient-echo, echo-planar images were acquired every 6 seconds during consecutive 20-minute trials. Nicotine induced a dose-dependent increase in several behavioral parameters, including feelings of "rush" and "high" and drug liking. Nicotine also induced a dose-dependent increase in neuronal activity in a distributed system of brain regions, including the nucleus accumbens, amygdala, cingulate, and frontal lobes. Activation in these structures is consistent with nicotine's behavior-arousing and behavior-reinforcing properties in humans. The identified brain regions have been previously shown to participate in the reinforcing, mood-elevating, and cognitive properties of other abused drugs such as cocaine, amphetamine, and opiates, suggesting that nicotine acts similarly in the human brain to produce its reinforcing and dependence properties.

Maternal cigarette smoking and child psychiatric morbidity: a longitudinal study.

Previous studies have linked maternal smoking during pregnancy with behavioral disturbance in children. However, additional evidence is needed to address the causality of the relationship. The present study analyses result from an Australian cohort of 5342 5-year-old children whose mothers were recruited early in pregnancy. Smoking history was gathered for prepregnancy, first clinic visit (FCV), late pregnancy, and when the child was 6 months and 5 years of age. Behavior problems at the age of 5 were assessed using a modified Child Behavior Check List (CBCL) shown to have high agreement with the complete CBCL. This resulted in the formation of three scales: internalizing; social, attentional and thought; and externalizing behavior problems, which were then dichotimized at the 90th percentile in each case. Logistic regression was used to model these outcomes as a function of maternal smoking at five time points during which it was assessed. A series of models explored the effect of additional adjustment for confounding. The predictors of attrition (29.5%) throughout the cohort were also identified by multivariate modeling. The final analysis was carried out on a cohort of mother-child pairs for whom data and child behavior outcomes were complete. The mean age of children was 5 years, 6 months with a range from 4 to 6 years. The mean age of mothers at the time of birth of the child was 25 years, with a range from 13 to 47 years. Mothers lost to follow-up were more likely to be younger, single, and less well-educated than those who continued participation, although maternal smoking was not an independent determinant. Unadjusted analyses showed strong associations between externalizing child behavior and maternal smoking during pregnancy and at the 5-year follow-up, with relative risks (RRs) up to 2.6 for children of women smoking at least 20 cigarettes per day at the first antenatal clinic visit. A clear dose-response relationship existed in most relationships with higher levels of smoking being associated with higher rates of externalizing behavior problems. Weaker relationships occurred for internalizing behavior and social, attentional and thought behavior problems. Multivariate analysis of the timing in more detail that the association between maternal smoking and child behavior problems persisted, although the evidence for dose-response diminished. Moreover, it was primarily associated with smoking as determined by questions asked at the FCV (RR = 1.52, 2.03, 2.16) for 1 to 9, 10 to 19, and >/=20 cigarettes per day, respectively, compared with nonsmoking and secondarily by smoking determined at the 5-year follow-up (RR = 1.52, 1.87, 1.29) for 1 to 9, 10 to 19, and >/=20 cigarettes per day respectively, compared with nonsmoking. This association appeared to be independent of a wide range of possible confounders such as maternal age, education, social class, marital status and mental health, gestation at FCV, complications during pregnancy, the child's sex, gestational age at birth, and age at last follow-up. Adjustments were also made for the mother's employment since birth, family structure, and maternal mental health at the time of the CBCL assessment. Associations between externalizing behavior problems and maternal smoking at other times, and those between other behavioral problems examined and maternal smoking were not significant. Although previous studies have found evidence for an association between maternal smoking and child behavior problems, the strength of this study lies in its size, its detailed and consistent measurement of maternal smoking, and its ability to control for many social and biological factors linked to maternal smoking and child behavior. (ABSTRACT TRUNCATED)