RESUMO
With the rise of e-cigarette use, teen nicotine exposure is becoming more widespread. Findings from clinical and preclinical studies show that the adolescent brain is particularly sensitive to nicotine. Animal studies have demonstrated that adolescent nicotine exposure increases reinforcement for cocaine and other drugs. However, the mechanisms that underlie these behaviors are poorly understood. Here, we report reactive microglia are critical regulators of nicotine-induced increases in adolescent cocaine self-administration. Nicotine has dichotomous, age-dependent effects on microglial morphology and immune transcript profiles. A multistep signaling mechanism involving D2 receptors and CX3CL1 mediates nicotine-induced increases in cocaine self-administration and microglial activation. Moreover, nicotine depletes presynaptic markers in a manner that is microglia-, D2- and CX3CL1-dependent. Taken together, we demonstrate that adolescent microglia are uniquely susceptible to perturbations by nicotine, necessary for nicotine-induced increases in cocaine-seeking, and that D2 receptors and CX3CL1 play a mechanistic role in these phenomena.
Assuntos
Cocaína/farmacologia , Comportamento de Procura de Droga/efeitos dos fármacos , Microglia/efeitos dos fármacos , Microglia/metabolismo , Nicotina/farmacologia , Aminopiridinas/farmacologia , Animais , Quimiocina CX3CL1/metabolismo , Modelos Animais de Doenças , Sistemas Eletrônicos de Liberação de Nicotina , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Masculino , Minociclina/farmacologia , Fenótipo , Pirróis/farmacologia , Ratos , Ratos Sprague-Dawley , Receptores de Dopamina D2/efeitos dos fármacos , Reforço Psicológico , Recompensa , Autoadministração , SinaptofisinaRESUMO
Addiction is a devastating disorder that produces persistent maladaptive changes to the central nervous system, including glial cells. Although there is an extensive body of literature examining the neuronal mechanisms of substance use disorders, effective therapies remain elusive. Glia, particularly microglia and astrocytes, have an emerging and meaningful role in a variety of processes beyond inflammation and immune surveillance, and may represent a promising therapeutic target. Indeed, glia actively modulate neurotransmission, synaptic connectivity and neural circuit function, and are critically poised to contribute to addictive-like brain states and behaviors. In this review, we argue that glia influence the cellular, molecular, and synaptic changes that occur in neurons following drug exposure, and that this cellular relationship is critically modified following drug exposure. We discuss direct actions of abused drugs on glial function through immune receptors, such as Toll-like receptor 4, as well as other mechanisms. We highlight how drugs of abuse affect glia-neural communication, and the profound effects that glial-derived factors have on neuronal excitability, structure, and function. Recent research demonstrates that glia have brain region-specific functions, and glia in different brain regions have distinct contributions to drug-associated behaviors. We will also evaluate the evidence demonstrating that glial activation is essential for drug reward and drug-induced dopamine release, and highlight clinical evidence showing that glial mechanisms contribute to drug abuse liability. In this review, we synthesize the extensive evidence that glia have a unique, pivotal, and underappreciated role in the development and maintenance of addiction.
Assuntos
Astrócitos/metabolismo , Encéfalo/metabolismo , Drogas Ilícitas/efeitos adversos , Microglia/metabolismo , Transtornos Relacionados ao Uso de Substâncias/metabolismo , Animais , Astrócitos/efeitos dos fármacos , Astrócitos/imunologia , Encéfalo/efeitos dos fármacos , Encéfalo/imunologia , Humanos , Microglia/efeitos dos fármacos , Microglia/imunologia , Neuroglia/efeitos dos fármacos , Neuroglia/imunologia , Neuroglia/metabolismo , Neurônios/efeitos dos fármacos , Neurônios/imunologia , Neurônios/metabolismo , Transtornos Relacionados ao Uso de Substâncias/imunologia , Receptor 4 Toll-Like/imunologia , Receptor 4 Toll-Like/metabolismoRESUMO
Myelination defects in the central nervous system (CNS) are associated with various psychiatric disorders, including drug addiction. As these disorders are often observed in individuals prenatally exposed to cigarette smoking, we tested the hypothesis that such exposure impairs central myelination in adolescence, an important period of brain development and the peak age of onset of psychiatric disorders. Pregnant Sprague Dawley rats were treated with nicotine (3 mg kg(-1) per day; gestational nicotine (GN)) or gestational saline via osmotic mini pumps from gestational days 4-18. Both male and female offsprings were killed on postnatal day 35 or 36, and three limbic brain regions, the prefrontal cortex (PFC), caudate putamen and nucleus accumbens, were removed for measurement of gene expression and determination of morphological changes using quantitative real-time PCR (qRT-PCR) array, western blotting and immunohistochemical staining. GN altered myelin gene expression at both the mRNA and protein levels, with striking sex differences. Aberrant expression of myelin-related transcription and trophic factors was seen in GN animals, which correlated highly with the alterations in the myelin gene expression. These correlations suggest that these factors contribute to GN-induced alterations in myelin gene expression and also indicate abnormal function of oligodendrocytes (OLGs), the myelin-producing cells in the CNS. It is unlikely that these changes are attributable solely to an alteration in the number of OLGs, as the cell number was changed only in the PFC of GN males. Together, our findings suggest that abnormal brain myelination underlies various psychiatric disorders and drug abuse associated with prenatal exposure to cigarette smoke.
Assuntos
Encéfalo/efeitos dos fármacos , Bainha de Mielina/metabolismo , Nicotina/farmacologia , Efeitos Tardios da Exposição Pré-Natal/induzido quimicamente , Animais , Western Blotting , Encéfalo/metabolismo , Química Encefálica/efeitos dos fármacos , Feminino , Expressão Gênica/efeitos dos fármacos , Masculino , Proteína Básica da Mielina/química , Proteína Básica da Mielina/metabolismo , Bainha de Mielina/química , Fibras Nervosas Mielinizadas/efeitos dos fármacos , Fibras Nervosas Mielinizadas/metabolismo , Córtex Pré-Frontal/química , Córtex Pré-Frontal/efeitos dos fármacos , Córtex Pré-Frontal/metabolismo , Gravidez , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Fatores Sexuais , Fatores de Transcrição/química , Fatores de Transcrição/metabolismoRESUMO
We have combined anatomical and functional methodologies to provide a comprehensive analysis of the properties of nicotinic acetylcholine receptors (nAChRs) on developing dopamine (DA) neurons of Sprague-Dawley rats. Double-labeling in situ hybridization was used to examine the expression of nAChR subunit mRNAs within developing midbrain DA neurons. As brain maturation progressed there was a change in the pattern of subunit mRNA expression within DA neurons, such that alpha3 and alpha4 subunits declined and alpha6 mRNA increased. Although there were strong similarities in subunit mRNA expression in substantia nigra (SNc) and ventral tegmental area (VTA), there was higher expression of alpha4 mRNA in SNc than VTA at gestational day (G) 15, and of alpha5, alpha6 and beta3 mRNAs during postnatal development. Using a superfusion neurotransmitter release paradigm to functionally characterize nicotine-stimulated release of [(3)H]DA from striatal slices, the properties of the nAChRs on DA terminals were also found to change with age. Functional nAChRs were detected on striatal terminals at G18. There was a decrease in maximal release in the first postnatal week, followed by an increase in nicotine efficacy and potency during the second and third postnatal weeks. In the transition from adolescence (postnatal days (P) 30 and 40) to adulthood, there was a complex pattern of functional maturation of nAChRs in ventral, but not dorsal, striatum. In males, but not females, there were significant changes in both nicotine potency and efficacy during this developmental period. These findings suggest that nAChRs may play critical functional roles throughout DA neuronal maturation.
Assuntos
Dopamina/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Neurônios/metabolismo , Receptores Nicotínicos/metabolismo , Substância Negra/metabolismo , Área Tegmentar Ventral/metabolismo , Envelhecimento/fisiologia , Animais , Animais Recém-Nascidos , Corpo Estriado/crescimento & desenvolvimento , Corpo Estriado/metabolismo , Feminino , Masculino , Neurônios/efeitos dos fármacos , Nicotina/farmacologia , Agonistas Nicotínicos/farmacologia , Gravidez , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Efeitos Tardios da Exposição Pré-Natal/fisiopatologia , Terminações Pré-Sinápticas/efeitos dos fármacos , Terminações Pré-Sinápticas/metabolismo , Subunidades Proteicas/genética , Subunidades Proteicas/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores Nicotínicos/genética , Caracteres Sexuais , Substância Negra/crescimento & desenvolvimento , Regulação para Cima/fisiologia , Área Tegmentar Ventral/crescimento & desenvolvimentoRESUMO
Gestational exposure of rats to nicotine produces long-lasting alterations in brain development. Vaccination of adult female rats against nicotine reduces the distribution of maternally administered nicotine to fetal brain, suggesting that vaccination might protect against these effects. In the current study, the effects of vaccination on nicotine-induced changes in fetal (3)H-epibatidine binding and c-fos mRNA expression were evaluated using tissue from a previous pharmacokinetic study of vaccination. An intermittent nicotine dosing regimen designed to resemble nicotine intake in a smoker was administered from GD1-20. Peak nicotine levels in fetal brain were reduced by vaccination, whereas the chronic accumulation of nicotine in fetal brain was not. Gestational nicotine exposure produced significant increases in (125)I-epibatidine binding to brain and spinal cord on GD20, and decreased c-fos mRNA expression in fetal striatum, adrenal and lung. Vaccination did not significantly alter these effects. These data suggest that nicotine dosing, using a clinically relevant intermittent bolus dose regimen, produces substantial changes in fetal nicotinic receptor and c-fos mRNA expression. The decrease in c-fos mRNA expression contrasts with previously reported increases, and suggests that the nicotine dosing regimen used may influence its effects. The lack of effect of vaccination suggests that the cumulative exposure of fetal tissues to nicotine may influence the measured parameters to a greater extent than peak exposure levels.
Assuntos
Expressão Gênica/fisiologia , Nicotina/imunologia , Nicotina/farmacologia , Proteínas Proto-Oncogênicas c-fos/metabolismo , Receptores Nicotínicos/metabolismo , Vacinação , Análise de Variância , Animais , Autorradiografia/métodos , Encéfalo/efeitos dos fármacos , Compostos Bicíclicos Heterocíclicos com Pontes/farmacocinética , Embrião de Mamíferos , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Expressão Gênica/efeitos dos fármacos , Agonistas Nicotínicos/farmacocinética , Gravidez , Ligação Proteica/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-fos/genética , Piridinas/farmacocinética , RNA Mensageiro/metabolismo , Ratos , Receptores Nicotínicos/efeitos dos fármacos , Trítio/farmacocinéticaRESUMO
Nicotinic acetylcholine receptor (nAChR)-evoked release of norepinephrine (NE) has been demonstrated in a number of brain regions that receive sole noradrenergic innervation from the locus coeruleus (LC). Many of these structures display enhanced nicotine-stimulated NE release in the neonate. We have examined the hypothalamus in order to determine if this region, which receives NE projections from both the LC and medullary catecholaminergic nuclei, also demonstrates maturational changes in nAChR-mediated NE release. Quantification of radiolabeled-NE release from rat hypothalamus slices by a maximally effective dose of nicotine revealed a peak response during the first postnatal week. This was followed by a decrease at postnatal day (P) 14, and a second peak at P21. Thereafter, release was equivalent to that observed at P14. Comparison of the pharmacological properties of nAChRs mediating NE release in neonatal (P7) and mature hypothalamus suggested involvement of different nAChR subtypes at the two ages. Using the selective toxin, DSP-4, nAChR-mediated NE release in the neonatal hypothalamus was shown to be from LC terminals. Our findings demonstrate an early sensitivity of hypothalamic LC terminals to nAChR regulation that may be associated with development of systems controlling critical homeostatic functions such as stress, feeding and cardiovascular regulation.
Assuntos
Animais Recém-Nascidos/metabolismo , Hipotálamo/metabolismo , Norepinefrina/metabolismo , Receptores Nicotínicos/metabolismo , Aconitina/análogos & derivados , Aconitina/farmacologia , Animais , Benzilaminas/farmacologia , Relação Dose-Resposta a Droga , Hipotálamo/citologia , Técnicas In Vitro , Masculino , Neurônios Aferentes/efeitos dos fármacos , Neurônios Aferentes/metabolismo , Neurotransmissores/metabolismo , Antagonistas Nicotínicos/farmacologia , Ratos , Ratos Sprague-Dawley , Receptores Nicotínicos/efeitos dos fármacos , Tubocurarina/farmacologiaRESUMO
The present study focuses on the nicotine-induced modulation of mRNA and protein expression of a number of genes involved in glutamatergic synaptic transmission in rat brain over different time periods of exposure. A subchronic (3 days) but not the chronic (7 or 14 days) administration of nicotine resulted in the up-regulation of Homer2a/b mRNA in the amygdala while in the ventral tegmental area (VTA) no change in expression of either Homer2a/b or Homer1b/c was observed. Although the increase in Homer2a/b mRNA was not translated into the protein level in the amygdala, a slight but significant up-regulation of Homer1b/c protein was observed in the same region at day 3. Both Homer forms were up-regulated at the protein level in the VTA at day 3. In the nucleus accumbens, 14 days of nicotine treatment up-regulated mRNA of Homer2b/c by 68.2% (P < 0.05), while the short form Homer1a gene was down-regulated by 65.0% at day 3 (P < 0.05). In regard to other components of the glutamatergic signalling, we identified an acute and intermittent increase in the mRNA and protein levels of mGluR1 and mGluR5 in the amygdala. In the VTA, however, the effects of nicotine on mGluR mRNA expression were long-lasting but rather specific to mGluR1. Nevertheless, mGluR1 protein levels in the VTA area were up-regulated only at day 3, as in the amygdala. These data provide further evidence for the involvement of nicotine in the glutamatergic neuronal synaptic activity in vivo, suggesting a role for the newly identified Homer proteins in this paradigm.
Assuntos
Proteínas de Transporte/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Nicotina/administração & dosagem , Agonistas Nicotínicos/administração & dosagem , Receptores de Glutamato Metabotrópico/metabolismo , Tonsila do Cerebelo/efeitos dos fármacos , Tonsila do Cerebelo/metabolismo , Animais , Western Blotting/métodos , Proteínas de Transporte/classificação , Proteínas de Transporte/genética , Esquema de Medicação , Expressão Gênica/efeitos dos fármacos , Proteínas de Arcabouço Homer , Masculino , Modelos Neurológicos , RNA Mensageiro/metabolismo , Ratos , Receptores de Glutamato Metabotrópico/classificação , Receptores de Glutamato Metabotrópico/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Fatores de Tempo , Área Tegmentar Ventral/efeitos dos fármacos , Área Tegmentar Ventral/metabolismoRESUMO
Nicotine enhances cognitive and attentional processes through stimulation of the basal forebrain cholinergic system. Although muscarinic cholinergic autoreceptors have been well characterized, pharmacological characterization of nicotinic autoreceptors has proven more difficult. The present study used double-labeling in situ hybridization to determine expression of nicotinic acetylcholine receptor (nAChR) subunit mRNAs within basal forebrain cholinergic neurons in order to gain information about possible nAChR autoreceptor properties. Cholinergic cells of the mesopontine tegmentum and striatal interneurons were also examined, as were septohippocampal GABAergic neurons that interact with cholinergic neurons to regulate hippocampal activity. alpha7 and beta2 nAChR mRNAs were found to be co-expressed in almost all cholinergic cells and in the majority of GABAergic neurons examined. alpha4 nAChR mRNA expression was restricted to cholinergic cells of the nucleus basalis magnocellularis, and to non-cholinergic cells of the medial septum and mesopontine tegmentum. These data suggest possible regional differences in the pharmacological properties of nicotinic autoreceptors on cholinergic cells. Whereas most cholinergic cells express rapidly desensitizing alpha7 homomers or alpha7beta2 heteromers, cortical projection neurons may also express a pharmacologically distinct alpha4beta2 nAChR subtype. There may also be differential nAChR regulation of cholinergic and non-cholinergic cells within the mesopontine tegmentum that are implicated in acquisition of nicotine self-administration.
Assuntos
Acetilcolina/metabolismo , Encéfalo/metabolismo , Fibras Colinérgicas/metabolismo , Neurônios/metabolismo , Receptores Nicotínicos/genética , Animais , Núcleo Basal de Meynert/citologia , Núcleo Basal de Meynert/metabolismo , Encéfalo/citologia , Fibras Colinérgicas/ultraestrutura , Regulação da Expressão Gênica/genética , Masculino , Neostriado/citologia , Neostriado/metabolismo , Neurônios/citologia , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Tegmento Mesencefálico/citologia , Tegmento Mesencefálico/metabolismo , Receptor Nicotínico de Acetilcolina alfa7 , Ácido gama-Aminobutírico/metabolismoRESUMO
Early neonatal handling of rat pups produces dampened hypothalamic-pituitary-adrenal axis reactivity to stress in adult male offspring. However, less is known about whether there is a similar effect for females. Although, most studies of neonatal handling have examined subsequent effects during adulthood, adolescence is an important developmental stage for stress responsivity. To address these issues, the effect of neonatal handling on the endocrine stress response and brain activity of male and female rats was determined in response to acute restraint stress during adolescence. Consistent with previous findings in adult males, neonatal handling reduced restraint stress-induced hormone levels in adolescent males. However, in contrast, we found elevated plasma hormone concentrations in handled females. A gender-specific handling effect on brain activity was also evident, with significantly increased stress-induced activation of the posterior cingulate cortex of handled females, as measured by c-fos mRNA expression. The striking gender difference in the effect of early neonatal handling provides evidence that this must be considered as an important variable in subsequent stress responsivity induced by early manipulations.
Assuntos
Caracteres Sexuais , Maturidade Sexual/fisiologia , Estresse Fisiológico/fisiopatologia , Doença Aguda , Hormônio Adrenocorticotrópico/sangue , Fatores Etários , Animais , Animais Recém-Nascidos , Corticosterona/sangue , Feminino , Giro do Cíngulo/crescimento & desenvolvimento , Giro do Cíngulo/fisiologia , Masculino , Gravidez , Proteínas Proto-Oncogênicas c-fos/genética , RNA Mensageiro/análise , Ratos , Restrição FísicaRESUMO
Presynaptic modulation of synaptic transmission is the primary function of central nicotinic acetylcholine receptors (nAChRs) in developing and adult brain. nAChR activation regulates release of various neurotransmitters, including norepinephrine (NA). Given evidence that NA may serve a critical functional role in cerebellar development, we have undertaken studies to determine whether nAChRs modulate NA release in developing cerebellum. In vitro experiments using cerebellar slices examined the effects of nAChR stimulation on release of radiolabeled NA ([3H]NA). Our data indicate the presence of functional nAChRs on NA terminals in immature cerebellum and subsequent developmental regulation of receptor properties. During postnatal week one, the maximally effective dose of nicotine released 35.0 +/- 1.2% of cerebellar [3H]NA stores. There was a subsequent decline in maximal nicotine-stimulated NA release until postnatal day 30, when Emax values were statistically indistinguishable from adult. Although the efficacy of nicotine changed substantially throughout development, EC50 values did not differ significantly (EC50 = 4.4-12.0 micro m). Pharmacological analysis indicated that this developmental shift in maximum nicotine effect reflects a change in the properties of the nAChRs. These data support recent findings of a possible functional role of nAChRs in regulating cerebellar ontogeny, and provides further support for the role of NA as a neurotrophic factor during development.
Assuntos
Cerebelo/crescimento & desenvolvimento , Cerebelo/metabolismo , Fluoxetina/análogos & derivados , Regulação da Expressão Gênica no Desenvolvimento , Norepinefrina/metabolismo , Receptores Nicotínicos/metabolismo , Fatores Etários , Envelhecimento/metabolismo , Animais , Animais Recém-Nascidos , Relação Dose-Resposta a Droga , Feminino , Fluoxetina/farmacologia , Técnicas In Vitro , Masculino , Nicotina/farmacologia , Agonistas Nicotínicos/farmacologia , Antagonistas Nicotínicos/farmacologia , Norepinefrina/farmacocinética , Proteínas da Membrana Plasmática de Transporte de Norepinefrina , Ratos , Ratos Sprague-Dawley , Receptores Nicotínicos/efeitos dos fármacos , Receptores Nicotínicos/genética , Simportadores/antagonistas & inibidores , TrítioRESUMO
Prolactin releasing peptide (PrRP) is a recently identified neuropeptide that stimulates prolactin release from pituitary cells. The presence of its receptor outside the hypothalamic-pituitary axis suggests that it may have other functions. We present here evidence that PrRP can modulate the activity of the reticular thalamic nucleus, a brain region with prominent PrRP receptor expression that is critical for sleep regulation and the formation of non-convulsive absence seizures. Intracerebroventricular injection of PrRP (1-10 nmol) into sleeping animals significantly suppresses sleep oscillations and promotes rapid and prolonged awakening. Higher concentrations of PrRP (10-100 nmol) similarly suppress spike wave discharges seen during absence seizures in genetic absence epilepsy rats from Strasbourg, an animal model for this disorder. In concordance with these findings, PrRP suppressed evoked oscillatory burst activity in reticular thalamic slices in vitro. These results indicate that PrRP modulates reticular thalamic function and that activation of its receptor provides a new target for therapies directed at sleep disorders and absence seizures.
Assuntos
Epilepsia Tipo Ausência/metabolismo , Hormônios Hipotalâmicos/metabolismo , Núcleos Intralaminares do Tálamo/metabolismo , Neurônios/metabolismo , Neuropeptídeos/metabolismo , Receptores de Neuropeptídeos/metabolismo , Transtornos do Sono-Vigília/metabolismo , Vigília/fisiologia , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/fisiologia , Animais , Relógios Biológicos/efeitos dos fármacos , Relógios Biológicos/fisiologia , Eletroencefalografia/efeitos dos fármacos , Epilepsia Tipo Ausência/tratamento farmacológico , Epilepsia Tipo Ausência/fisiopatologia , Glutamato Descarboxilase/genética , Glutamato Descarboxilase/metabolismo , Hormônios Hipotalâmicos/farmacologia , Núcleos Intralaminares do Tálamo/efeitos dos fármacos , Núcleos Intralaminares do Tálamo/fisiopatologia , Isoenzimas/genética , Isoenzimas/metabolismo , Masculino , Neurônios/efeitos dos fármacos , Neuropeptídeos/farmacologia , Técnicas de Cultura de Órgãos , Hormônio Liberador de Prolactina , RNA Mensageiro/metabolismo , Ratos , Ratos Mutantes , Ratos Sprague-Dawley , Receptores de Neuropeptídeos/efeitos dos fármacos , Receptores de Neuropeptídeos/genética , Sono/efeitos dos fármacos , Sono/fisiologia , Transtornos do Sono-Vigília/tratamento farmacológico , Transtornos do Sono-Vigília/fisiopatologia , Vigília/efeitos dos fármacos , Ácido gama-Aminobutírico/biossínteseRESUMO
The endogenous cannabinoids (endocannabinoids) are lipid molecules that may mediate retrograde signaling at central synapses and other forms of short-range neuronal communication. The monoglyceride 2-arachidonoylglycerol (2-AG) meets several criteria of an endocannabinoid substance: (i) it activates cannabinoid receptors; (ii) it is produced by neurons in an activity-dependent manner; and (iii) it is rapidly eliminated. 2-AG inactivation is only partially understood, but it may occur by transport into cells and enzymatic hydrolysis. Here we tested the hypothesis that monoglyceride lipase (MGL), a serine hydrolase that converts monoglycerides to fatty acid and glycerol, participates in 2-AG inactivation. We cloned MGL by homology from a rat brain cDNA library. Its cDNA sequence encoded for a 303-aa protein with a calculated molecular weight of 33,367 daltons. Northern blot and in situ hybridization analyses revealed that MGL mRNA is heterogeneously expressed in the rat brain, with highest levels in regions where CB(1) cannabinoid receptors are also present (hippocampus, cortex, anterior thalamus, and cerebellum). Immunohistochemical studies in the hippocampus showed that MGL distribution has striking laminar specificity, suggesting a presynaptic localization of the enzyme. Adenovirus-mediated transfer of MGL cDNA into rat cortical neurons increased MGL expression and attenuated N-methyl-D-aspartate/carbachol-induced 2-AG accumulation in these cells. No such effect was observed on the accumulation of anandamide, another endocannabinoid lipid. The results suggest that hydrolysis by means of MGL is a primary mechanism for 2-AG inactivation in intact neurons.
Assuntos
Encéfalo/enzimologia , Glicerídeos/metabolismo , Monoacilglicerol Lipases/metabolismo , Sequência de Aminoácidos , Animais , Ácidos Araquidônicos/metabolismo , Sequência de Bases , Encéfalo/citologia , Células COS , Moduladores de Receptores de Canabinoides , Canabinoides/metabolismo , Células Cultivadas , Chlorocebus aethiops , DNA Complementar , Endocanabinoides , Expressão Gênica , Células HeLa , Humanos , Hidrólise , Dados de Sequência Molecular , Monoacilglicerol Lipases/genética , Neurônios/citologia , Neurônios/enzimologia , Alcamidas Poli-Insaturadas , Ratos , Ratos WistarRESUMO
The primary role of nicotinic acetylcholine receptors in adult and developing brain is to modulate neurotransmission. Using in vitro neurotransmitter release, we have examined mechanisms underlying nicotine-induced [(3)H]norepinephrine release from developing and adult rat hippocampus. At birth, nicotine significantly stimulated hippocampal [(3)H]norepinephrine release with a monotonic increase in maximal drug effect over the first ten postnatal days. No developmental changes in agonist or antagonist potency were observed. Comparison of synaptosomal and slice preparations, as well as examination of the effects of tetrodotoxin, indicated that at least two nicotinic acetylcholine receptor populations regulated [(3)H]norepinephrine release from neonatal and adult hippocampus; one localized on noradrenergic terminals, the other on adjacent cells. To further characterize the indirect mechanism of nicotine action in the adult, we examined the effects of pharmacological blockade of various neurotransmitter systems that provide excitatory input to hippocampal noradrenergic terminals. Whereas glutamate and muscarinic receptor blockade was ineffective, the GABA-A receptor antagonists, bicuculline and picrotoxin, inhibited the indirect component of nicotine-mediated [(3)H]norepinephrine release. Furthermore, pentobarbital, an allosteric effector at GABA-A receptors, potentiated the effect of submaximal concentrations of nicotine. These findings are consistent with the hypothesis that nicotine-induced GABA release serves as an additional stimulus for [(3)H]norepinephrine secretion within rat hippocampus.
Assuntos
Hipocampo/crescimento & desenvolvimento , Hipocampo/metabolismo , Norepinefrina/metabolismo , Receptores Nicotínicos/fisiologia , Animais , Animais Recém-Nascidos , Relação Dose-Resposta a Droga , Feminino , Hipocampo/efeitos dos fármacos , Masculino , Nicotina/farmacologia , Agonistas Nicotínicos/farmacologia , Antagonistas Nicotínicos/farmacologia , Gravidez , Ratos , Ratos Sprague-Dawley , TrítioRESUMO
Urotensin II (UII) is a peptide known to be a potent vasoconstrictor. The urotensin II receptor (UII-R) is expressed not only in peripheral tissues but also in the brain of rodents. As a basis for studies of UII central nervous system actions, UII-R localization in the rat brain was analyzed by in situ hybridization and by in situ binding. UII-R mRNA was found in the mesopontine tegmental area colocalizing with choline acetyltransferase. Binding sites were detected throughout the brain with the highest levels found in the pedunculopontine tegmental area, the lateral dorsal tegmental area, and the lateral septal, medial habenular, and interpeduncular nuclei. The majority of these brain nuclei are sites of axonal termination originating from the mesopontine areas, suggesting that UII-R is a presynaptic receptor. This distribution of UII-R in the cholinergic mesopontine area indicates that the UII system may be involved in sensory-motor integration and perhaps in central nervous system blood flow.
Assuntos
Acetilcolina/fisiologia , Ponte/fisiologia , Receptores de Superfície Celular/genética , Receptores Acoplados a Proteínas G , Tegmento Mesencefálico/fisiologia , Animais , Colina O-Acetiltransferase/genética , Expressão Gênica/fisiologia , Habenula/fisiologia , Hibridização In Situ , Radioisótopos do Iodo , Masculino , RNA Mensageiro/análise , Ratos , Ratos Sprague-Dawley , Receptores Pré-Sinápticos/fisiologia , Núcleos Septais/fisiologiaRESUMO
The melanin-concentrating hormone (MCH) system is thought to be an important regulator of food intake. Recently the orphan G protein-coupled receptor SLC-1 was identified as the MCH receptor (MCHR). Preliminary analyses of MCHR mRNA distribution have supported a role for the MCH system in nutritional homeostasis. We report here a complete anatomical distribution of the MCHR mRNA. We have found high levels of expression of MCHR mRNA in most anatomical areas implicated in control of olfaction, with the exception of the main olfactory bulb. Dense labeling was also detected in the hippocampal formation, subiculum, and basolateral amygdala, all of which are important in learning and memory, and in the shell of the nucleus accumbens, a substrate for motivated behavior and feeding. Within the hypothalamus, MCHR mRNA was moderately expressed in the ventromedial nucleus, arcuate nucleus, and zona incerta, all of which serve key roles in the neuronal circuitry of feeding. In the brainstem, strong expression was observed in the locus coeruleus, which is implicated in arousal, as well as in nuclei that contribute to orofacial function and mastication, including the facial, hypoglossal, motor trigeminal, and dorsal motor vagus nuclei. In most regions there was a good correspondence between MCHR mRNA distribution and that of MCH-immunoreactive fibers. Taken together, these data suggest that MCH may act at various levels of the brain to integrate various aspects of feeding behavior. However, the extensive MCHR distribution throughout the brain suggests that this receptor may play a role in other functions, most notably reinforcement, arousal, sensorimotor integration, and autonomic control.
Assuntos
Hipocampo/fisiologia , Condutos Olfatórios/fisiologia , Ratos Sprague-Dawley/fisiologia , Receptores do Hormônio Hipofisário/genética , Tonsila do Cerebelo/química , Tonsila do Cerebelo/fisiologia , Animais , Apetite/fisiologia , Núcleo Arqueado do Hipotálamo/química , Núcleo Arqueado do Hipotálamo/fisiologia , Tronco Encefálico/química , Tronco Encefálico/fisiologia , Condicionamento Psicológico/fisiologia , Expressão Gênica/fisiologia , Hipocampo/química , Hibridização In Situ , Masculino , Núcleo Accumbens/química , Núcleo Accumbens/fisiologia , Condutos Olfatórios/química , RNA Mensageiro/análise , Ratos , Olfato/fisiologia , Núcleo Hipotalâmico Ventromedial/química , Núcleo Hipotalâmico Ventromedial/fisiologiaRESUMO
Cholinergic markers in the middle layers of rat auditory cortex are transiently upregulated during the second postnatal week, at which time alpha 7 nicotinic acetylcholine receptors (nAChRs) selectively regulate NMDA receptor (NMDAR)-mediated EPSPs. To investigate the developmental role of this regulation, we determined whether manipulating nAChR function at specific times during the first 4 weeks after birth could alter subsequent neuronal function. Rat pups were injected twice daily with nicotine (1 or 2 mg/kg) or saline during approximately the first, second, or fourth postnatal week (i. e., before, during, or after the peak upregulation of nAChRs). Glutamate EPSPs and intrinsic membrane properties were measured during whole-cell recordings from visually identified pyramidal neurons in layers II-IV of brain slices prepared at least 15 hr after the last injection. Chronic nicotine exposure (CNE) had little effect on intrinsic membrane properties and during week 1 or 4 did not affect synaptic function. However, CNE during week 2 resulted in EPSPs with long durations, multiple peaks, and enhanced NMDAR components. These changes remained significant even 10 d after CNE. Rapid application of nicotine, which in control neurons selectively enhances NMDAR EPSPs during week 2, produced only weak effects after CNE. Receptor binding studies showed that CNE-induced EPSP alterations occurred in the absence of altered alpha 7 nAChR numbers or agonist binding affinity. Thus, altered stimulation of nAChRs by CNE during week 2, but not before or after, disrupts the development of glutamate synapses in rat auditory cortex.
Assuntos
Córtex Auditivo/efeitos dos fármacos , Córtex Auditivo/crescimento & desenvolvimento , Neurônios/efeitos dos fármacos , Nicotina/toxicidade , Efeitos Tardios da Exposição Pré-Natal , Receptores Nicotínicos/efeitos dos fármacos , Sinapses/efeitos dos fármacos , Fatores Etários , Animais , Animais Recém-Nascidos , Córtex Auditivo/fisiopatologia , Esquema de Medicação , Potenciais Pós-Sinápticos Excitadores/efeitos dos fármacos , Potenciais Pós-Sinápticos Excitadores/fisiologia , Feminino , Potenciais da Membrana/efeitos dos fármacos , Potenciais da Membrana/fisiologia , Microinjeções , Neurônios/metabolismo , Neurônios/ultraestrutura , Gravidez , Ratos , Ratos Sprague-Dawley , Receptores de N-Metil-D-Aspartato/efeitos dos fármacos , Receptores de N-Metil-D-Aspartato/fisiologia , Receptores Nicotínicos/metabolismo , Sinapses/metabolismo , Sinapses/ultraestrutura , Transmissão Sináptica/efeitos dos fármacos , Transmissão Sináptica/fisiologia , Receptor Nicotínico de Acetilcolina alfa7RESUMO
A growing body of evidence indicates that neuronal nicotinic acetylcholine receptors (nAChRs), in addition to promoting fast cholinergic transmission, may modulate other neuronal activities within the central nervous system (CNS). In particular, the alpha7 nAChR is highly permeable to Ca2+ and may serve a distinct role in regulating neuronal plasticity. By elevating intracellular Ca2+ levels in discrete neuronal locations, these ligand-gated ion channels may influence numerous physiological processes in developing and adult CNS. In this article, we review evidence that both pre- and postsynaptic alpha7 nAChRs modulate transmitter release in the brain and periphery through Ca2+-dependent mechanisms. The possible role of alpha7 nAChRs in regulating neuronal growth and differentiation in developing CNS is also evaluated. We consider an interaction between cholinergic and glutamatergic transmission and propose a hypothesis on the possible coregulation of intracellular Ca2+ by N-methyl-D-aspartate (NMDA) receptors and alpha7 nAChRs. Finally, the clinical significance of alterations in the normal function of alpha7 nAChRs is discussed as it pertains to prenatal nicotine exposure, schizophrenia, and epilepsy.
Assuntos
Sistema Nervoso Central/fisiologia , Plasticidade Neuronal/fisiologia , Neurônios/fisiologia , Receptores Nicotínicos/fisiologia , Animais , Cálcio/fisiologia , Modelos Neurológicos , Transmissão Sináptica , Receptor Nicotínico de Acetilcolina alfa7RESUMO
To investigate the hypothesis that differing mRNA levels underlie gender differences in the contractile response of the rat tail artery, alpha2-adrenoceptor mRNA was measured using in situ hybridization. Messenger RNA for the alpha2A- and alpha2C-adrenoceptor subtypes was found localized to the smooth muscle layer. There was no detectable mRNA present for the alpha2B-adrenoceptor subtype. Levels of alpha2C-adrenoceptor mRNA were greater in female compared to male tail arteries (417 +/- 35 vs. 263 +/- 38 dpm/mg, P = 0.01), while levels of alpha2A-adrenoceptor mRNA were the same in both sexes. Levels of alpha2-adrenoceptor mRNA may parallel levels of functioning protein present in the rat tail artery.