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1.
Transl Psychiatry ; 13(1): 118, 2023 04 08.
Artigo em Inglês | MEDLINE | ID: mdl-37031222

RESUMO

The frontal pole (Brodmann area 10, BA10) is the largest cytoarchitectonic region of the human cortex, performing complex integrative functions. BA10 undergoes intensive adolescent grey matter pruning prior to the age of onset for bipolar disorder (BP) and schizophrenia (SCHIZ), and its dysfunction is likely to underly aspects of their shared symptomology. In this study, we investigated the role of BA10 neurotransmission-related gene expression in BP and SCHIZ. We performed qPCR to measure the expression of 115 neurotransmission-related targets in control, BP, and SCHIZ postmortem samples (n = 72). We chose this method for its high sensitivity to detect low-level expression. We then strengthened our findings by performing a meta-analysis of publicly released BA10 microarray data (n = 101) and identified sources of convergence with our qPCR results. To improve interpretation, we leveraged the unusually large database of clinical metadata accompanying our samples to explore the relationship between BA10 gene expression, therapeutics, substances of abuse, and symptom profiles, and validated these findings with publicly available datasets. Using these convergent sources of evidence, we identified 20 neurotransmission-related genes that were differentially expressed in BP and SCHIZ in BA10. These results included a large diagnosis-related decrease in two important therapeutic targets with low levels of expression, HTR2B and DRD4, as well as other findings related to dopaminergic, GABAergic and astrocytic function. We also observed that therapeutics may produce a differential expression that opposes diagnosis effects. In contrast, substances of abuse showed similar effects on BA10 gene expression as BP and SCHIZ, potentially amplifying diagnosis-related dysregulation.


Assuntos
Transtorno Bipolar , Esquizofrenia , Humanos , Adolescente , Transtorno Bipolar/genética , Transtorno Bipolar/metabolismo , Esquizofrenia/metabolismo , Lobo Frontal/metabolismo , Expressão Gênica , Transmissão Sináptica/genética
2.
Brain Struct Funct ; 226(2): 481-499, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33386994

RESUMO

Transcript labeling in intact tissues using in situ hybridization chain reaction has potential to provide vital spatiotemporal information for molecular characterization of heterogeneous neuronal populations. However, large tissue labeling in non-perfused or fresh-frozen rodent and postmortem human samples, which provide more flexible utilization than perfused tissues, is largely unexplored. In the present study, we optimized the combination of in situ hybridization chain reaction in fresh-frozen rodent brains and then evaluated the uniformity of neuronal labeling between two clearing methods, CLARITY and iDISCO+. We found that CLARITY yielded higher signal-to-noise ratios but more limited imaging depth and required longer clearing times, whereas, iDISCO+ resulted in better tissue clearing, greater imaging depth and a more uniform labeling of larger samples. Based on these results, we used iDISCO+-cleared fresh-frozen rodent brains to further validate this combination and map the expression of a few genes of interest pertaining to mood disorders. We then examined the potential of in situ hybridization chain reaction to label transcripts in cleared postmortem human brain tissues. The combination failed to produce adequate mRNA labeling in postmortem human cortical slices but produced visually adequate labeling in the cerebellum tissues. We next, investigated the multiplexing ability of in situ hybridization chain reaction in cleared tissues which revealed inconsistent fluorescence output depending upon the fluorophore conjugated to the hairpins. Finally, we applied our optimized protocol to assess the effect of glucocorticoid receptor overexpression on basal somatostatin expression in the mouse cortex. The constitutive glucocorticoid receptor overexpression resulted in lower number density of somatostatin-expressing neurons compared to wild type. Overall, the combination of in situ hybridization chain reaction with clearing methods, especially iDISCO+, may find broad application in the transcript analysis in rodent studies, but its limited use in postmortem human tissues can be improved by further optimizations.


Assuntos
Encéfalo/metabolismo , Hibridização in Situ Fluorescente , Neurônios/metabolismo , Animais , Humanos , Camundongos , Ratos , Ratos Sprague-Dawley
3.
Proc Natl Acad Sci U S A ; 115(40): E9489-E9498, 2018 10 02.
Artigo em Inglês | MEDLINE | ID: mdl-30224492

RESUMO

Two classes of peptide-producing neurons in the arcuate nucleus (Arc) of the hypothalamus are known to exert opposing actions on feeding: the anorexigenic neurons that express proopiomelanocortin (POMC) and the orexigenic neurons that express agouti-related protein (AgRP) and neuropeptide Y (NPY). These neurons are thought to arise from a common embryonic progenitor, but our anatomical and functional understanding of the interplay of these two peptidergic systems that contribute to the control of feeding remains incomplete. The present study uses a combination of optogenetic stimulation with viral and transgenic approaches, coupled with neural activity mapping and brain transparency visualization to demonstrate the following: (i) selective activation of Arc POMC neurons inhibits food consumption rapidly in unsated animals; (ii) activation of Arc neurons arising from POMC-expressing progenitors, including POMC and a subset of AgRP neurons, triggers robust feeding behavior, even in the face of satiety signals from POMC neurons; (iii) the opposing effects on food intake are associated with distinct neuronal projection and activation patterns of adult hypothalamic POMC neurons versus Arc neurons derived from POMC-expressing lineages; and (iv) the increased food intake following the activation of orexigenic neurons derived from POMC-expressing progenitors engages an extensive neural network that involves the endogenous opioid system. Together, these findings shed further light on the dynamic balance between two peptidergic systems in the moment-to-moment regulation of feeding behavior.


Assuntos
Proteína Agouti Sinalizadora/biossíntese , Núcleo Arqueado do Hipotálamo/metabolismo , Comportamento Alimentar/fisiologia , Neurônios/metabolismo , Neuropeptídeo Y/biossíntese , Pró-Opiomelanocortina/biossíntese , Transdução de Sinais/fisiologia , Proteína Agouti Sinalizadora/genética , Animais , Núcleo Arqueado do Hipotálamo/citologia , Comportamento Alimentar/psicologia , Camundongos , Camundongos Transgênicos , Neurônios/citologia , Neuropeptídeo Y/genética , Pró-Opiomelanocortina/genética
4.
J Comp Neurol ; 524(10): 2059-79, 2016 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-26587737

RESUMO

In the retina, dopamine is a key molecule for daytime vision. Dopamine is released by retinal dopaminergic amacrine cells and transmits signaling either by conventional synaptic or by volume transmission. By means of volume transmission, dopamine modulates all layers of retinal neurons; however, it is not well understood how dopamine modulates visual signaling pathways in bipolar cells. Here we analyzed Drd1a-tdTomato BAC transgenic mice and found that the dopamine D1 receptor (D1R) is expressed in retinal bipolar cells in a type-dependent manner. Strong tdTomato fluorescence was detected in the inner nuclear layer and localized to type 1, 3b, and 4 OFF bipolar cells and type 5-2, XBC, 6, and 7 ON bipolar cells. In contrast, type 2, 3a, 5-1, 9, and rod bipolar cells did not express Drd1a-tdTomato. Other interneurons were also found to express tdTomato including horizontal cells and a subset (25%) of AII amacrine cells. Diverse visual processing pathways, such as color or motion-coded pathways, are thought to be initiated in retinal bipolar cells. Our results indicate that dopamine sculpts bipolar cell performance in a type-dependent manner to facilitate daytime vision. J. Comp. Neurol. 524:2059-2079, 2016. © 2015 Wiley Periodicals, Inc.


Assuntos
Receptores de Dopamina D1/metabolismo , Retina/citologia , Células Bipolares da Retina/classificação , Células Bipolares da Retina/metabolismo , Vias Visuais/fisiologia , Células Amácrinas/metabolismo , Animais , Biotina/análogos & derivados , Biotina/metabolismo , Colina O-Acetiltransferase/metabolismo , Canais Disparados por Nucleotídeos Cíclicos Ativados por Hiperpolarização/metabolismo , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Proteínas do Tecido Nervoso/metabolismo , Opsinas/metabolismo , Canais de Potássio/metabolismo , RNA Mensageiro/metabolismo , Receptores de Dopamina D1/genética , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Células Bipolares da Retina/citologia , Sinaptotagmina II/metabolismo , Fatores de Transcrição/metabolismo
5.
J Comp Neurol ; 518(22): 4591-611, 2010 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-20886624

RESUMO

The hypothalamus regulates numerous autonomic responses and behaviors. The neuroactive substances corticotropin-releasing factor (CRF), arginine-vasopressin (AVP), histidine decarboxylase (HDC), melanin-concentrating hormone (MCH), and orexin/hypocretins (ORX) produced in the hypothalamus mediate a subset of these processes. Although the expression patterns of these genes have been well studied in rodents, less is known about them in humans. We combined classical histological techniques with in situ hybridization histochemistry to produce both 2D and 3D images and to visually align and quantify expression of the genes for these substances in nuclei of the human hypothalamus. The hypothalamus was arbitrarily divided into rostral, intermediate, and caudal regions. The rostral region, containing the paraventricular nucleus (PVN), was defined by discrete localization of CRF- and AVP-expressing neurons, whereas distinct relationships between HDC, MCH, and ORX mRNA-expressing neurons delineated specific levels within the intermediate and caudal regions. Quantitative mRNA signal intensity measurements revealed no significant differences in overall CRF or AVP expression at any rostrocaudal level of the PVN. HDC mRNA expression was highest at the level of the premammillary area, which included the dorsomedial and tuberomammillary nuclei as well as the dorsolateral hypothalamic area. In addition, the overall intensity of hybridization signal exhibited by both MCH and ORX mRNA-expressing neurons peaked in distinct intermediate and caudal hypothalamic regions. These results suggest that human hypothalamic neurons involved in the regulation of the HPA axis display distinct neurochemical patterns that may encompass multiple local nuclei.


Assuntos
Arginina Vasopressina/metabolismo , Hormônio Liberador da Corticotropina/metabolismo , Regulação da Expressão Gênica/fisiologia , Histidina Descarboxilase/metabolismo , Hormônios Hipotalâmicos/metabolismo , Hipotálamo/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Melaninas/metabolismo , Neuropeptídeos/metabolismo , Hormônios Hipofisários/metabolismo , Adulto , Idoso , Arginina Vasopressina/genética , Mapeamento Encefálico , Hormônio Liberador da Corticotropina/genética , Feminino , Perfilação da Expressão Gênica , Histidina Descarboxilase/genética , Humanos , Hormônios Hipotalâmicos/genética , Hipotálamo/citologia , Imageamento Tridimensional/métodos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Masculino , Melaninas/genética , Pessoa de Meia-Idade , Neurônios/metabolismo , Neuropeptídeos/genética , Orexinas , Hormônios Hipofisários/genética , RNA Mensageiro/metabolismo
6.
J Neurosci Res ; 87(7): 1645-58, 2009 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-19115412

RESUMO

Clinical and experimental studies implicate the use of serotonin (5-HT)1A receptor agonists for the reduction of L-3,4-dihydroxyphenylalanine (L-DOPA)-induced dyskinesia (LID). Although raphe nuclei likely play a role in these antidyskinetic effects, an unexplored population of striatal 5-HT1A receptors (5-HT1AR) may also contribute. To better characterize this mechanism, L-DOPA-primed hemiparkinsonian rats received the 5-HT1AR agonist +/-8-OH-DPAT (0, 0.1, 1.0 mg/kg, i.p.) with or without cotreatment with the 5-HT1AR antagonist WAY100635 (0.5 mg/kg, i.p.) 5 min after L-DOPA, after which abnormal involuntary movements (AIMs), rotations, and forelimb akinesia were quantified. To establish the effects of 5-HT1AR stimulation on L-DOPA-induced c-fos and preprodynorphin (PPD) mRNA within the dopamine-depleted striatum, immunohistochemistry and real-time reverse transcription polymerase chain reaction, respectively, were used. Finally, to determine the contribution of striatal 5-HT1AR to these effects, L-DOPA-primed hemiparkinsonian rats received bilateral intrastriatal microinfusions of +/-8-OH-DPAT (0, 5, or 10 microg/side), WAY100635 (5 microg/side), or both (10 microg + 5 microg/side) 5 min after L-DOPA, after which AIMs and rotations were examined. Systemic +/-8-OH-DPAT dose- and receptor-dependently attenuated L-DOPA-mediated AIMs and improved forelimb akinesia. Striatal c-fos immunoreactivity and PPD mRNA ipsilateral to the lesion were strongly induced by L-DOPA, while +/-8-OH-DPAT suppressed these effects. Finally, intrastriatal infusions of +/-8-OH-DPAT reduced AIMs while coinfusion of WAY100635 reversed its antidyskinetic effect. Collectively, these results support the hypothesis that the cellular and behavioral properties of 5-HT1AR agonists are conveyed in part via a population of functional 5-HT1AR within the striatum.


Assuntos
Antiparkinsonianos/uso terapêutico , Corpo Estriado/efeitos dos fármacos , Levodopa/uso terapêutico , Transtornos Parkinsonianos/tratamento farmacológico , Receptor 5-HT1A de Serotonina/metabolismo , 8-Hidroxi-2-(di-n-propilamino)tetralina/farmacologia , Animais , Dinorfinas/metabolismo , Discinesia Induzida por Medicamentos/tratamento farmacológico , Discinesia Induzida por Medicamentos/metabolismo , Levodopa/efeitos adversos , Masculino , Atividade Motora/efeitos dos fármacos , Oxidopamina , Transtornos Parkinsonianos/induzido quimicamente , Transtornos Parkinsonianos/metabolismo , Piperazinas/farmacologia , Precursores de Proteínas/metabolismo , Proteínas Proto-Oncogênicas c-fos/metabolismo , Piridinas/farmacologia , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Agonistas do Receptor 5-HT1 de Serotonina , Antagonistas do Receptor 5-HT1 de Serotonina , Antagonistas da Serotonina/farmacologia , Agonistas do Receptor de Serotonina/farmacologia
7.
Synapse ; 57(1): 1-7, 2005 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-15858838

RESUMO

Recent evidence suggests that striatal neurokinin receptors modulate dopamine (DA)-induced motor behaviors. To further examine this, we studied the effects of intrastriatal neurokinin 1 receptor (NK1R) antagonism on motor behaviors induced by direct infusion of the full DA D1 receptor agonist SKF 82958. Adult male Sprague-Dawley rats received bilateral intrastriatal 0.8-mul infusions of the NK1R receptor antagonist LY 306,740 (0, 27, or 54 nmol/side) followed by intrastriatal infusions of SKF 82958 (0 or 24 nmol/side) into the dorsal striatum. Following each infusion, rats were placed into automated activity monitors for the quantification of horizontal activity, total distance traveled, movement bouts, and stereotypy counts. As expected, SKF 82958 increased motor activity on all behavioral measures. More importantly, whereas 27 nmol was without effect, prior infusion of 54 nmol LY 306,740 significantly reduced most aspects of behavior. The results of this study suggest that functional NK1Rs within the striatum play a permissive role in the motor behaviors induced by D1R stimulation.


Assuntos
Agonistas de Dopamina/farmacologia , Atividade Motora/efeitos dos fármacos , Neostriado/metabolismo , Antagonistas dos Receptores de Neurocinina-1 , Receptores de Dopamina D1/agonistas , Acetamidas/administração & dosagem , Acetamidas/farmacologia , Animais , Benzazepinas/administração & dosagem , Benzazepinas/farmacologia , Agonistas de Dopamina/administração & dosagem , Masculino , Microinjeções , Neostriado/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Comportamento Estereotipado/efeitos dos fármacos
8.
Am J Psychiatry ; 160(1): 47-55, 2003 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-12505801

RESUMO

OBJECTIVE: The hypothesis that human cocaine users lose vesicular monoamine transporter (VMAT2) protein was tested in striatal samples from cocaine users and age-, sex-, and postmortem interval-matched comparison subjects. METHOD: Striatal samples were retrieved at autopsy; immunoblot assays were then performed by using a highly specific VMAT2 antibody. Striatal radioligand binding to VMAT2 was assessed with dihydrotetrabenazine ([(3)H]DTBZ) and dopamine levels employing high-performance liquid chromatography. RESULTS: Cocaine users displayed a marked reduction in VMAT2 immunoreactivity as well as reduced [(3)H]DTBZ binding and dopamine levels. It did not appear that the reduction in VMAT2 immunoreactivity was related to ethanol use, but dopamine levels were lower in subjects with only ethanol diagnoses. Subjects suffering from cocaine-induced mood disorders displayed a greater loss of VMAT2 immunoreactivity that approached significance. CONCLUSIONS: Human cocaine users lose VMAT2 protein, which might reflect damage to striatal dopamine fibers. These neuronal changes could play a role in causing disordered mood and motivational processes in more severely dependent patients.


Assuntos
Transtornos Relacionados ao Uso de Cocaína/genética , Corpo Estriado/patologia , Perda de Heterozigosidade , Glicoproteínas de Membrana/genética , Proteínas de Membrana Transportadoras , Neuropeptídeos , Adolescente , Adulto , Alcoolismo/genética , Alcoolismo/patologia , Western Blotting , Transtornos Relacionados ao Uso de Cocaína/patologia , Dopamina/metabolismo , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Valores de Referência , Proteínas Vesiculares de Transporte de Aminas Biogênicas , Proteínas Vesiculares de Transporte de Monoamina
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