RESUMO
OBJECTIVE: To establish in an exploratory neuroimaging study whether γ-hydroxybutyrate (sodium oxybate [SO]), a sedative, anti-narcoleptic drug with abuse potential, transiently inhibits striatal dopamine release in the human. METHODS: Ten healthy participants (30 years; 6M, 4F) and one participant with narcolepsy received a baseline positron emission tomography scan of [C-11]raclopride, a D2/3 dopamine receptor radioligand sensitive to dopamine occupancy, followed approximately one week later by an oral sedative 3g dose of SO and two [C-11]raclopride scans (1 h, 7 h post SO). Plasma SO levels and drowsiness duration were assessed. RESULTS: No significant changes were detected in [C-11]raclopride binding in striatum overall 1 or 7 h after SO, but a small non-significant increase in [C-11]raclopride binding, implying decreased dopamine occupancy, was noted in limbic striatal subdivision at one hour (+6.5%; p uncorrected = 0.045; +13.2%, narcolepsy participant), returning to baseline at 7 h. A positive correlation was observed between drowsiness duration and percent change in [C-11]raclopride binding in limbic striatum (r = 0.73; p = 0.017). CONCLUSIONS: We did not find evidence in this sample of human subjects of a robust striatal dopamine change, as was reported in non-human primates. Our preliminary data, requiring extension, suggest that a 3g sedative SO dose might cause slight transient inhibition of dopamine release in limbic striatum.
Assuntos
Dopamina , Oxibato de Sódio , Encéfalo/diagnóstico por imagem , Encéfalo/metabolismo , Radioisótopos de Carbono/metabolismo , Corpo Estriado/metabolismo , Dopamina/metabolismo , Humanos , Neuroimagem , Oxibato de Sódio/farmacologiaRESUMO
OBJECTIVES: Transient receptor potential canonical type 3 (TRPC3) channels are activated in B lymphoblast cell lines from patients with bipolar disorder (BD), and its expression is reduced by chronic lithium treatment, implicating TRPC3 in the intracellular calcium (Ca2+ ) dyshomeostasis of BD. Thrombin, via a protease-activated receptor, moderates Ca2+ signaling and TRPC3 in astrocytes, and also cell proliferation. We examined whether lithium pretreatment attenuates thrombin-stimulated TRPC3 expression and function in astrocytes, and levels of the calcium-binding peptide, S100B, which is expressed mainly in these cells. METHODS: Human astroglioma, U-87MG, cells were pretreated with 1 mmol L-1 LiCl for 1 day (acute), 3 days (subacute), and 7 days (chronic). To examine the role of TRPC3, genetically stable knockdown TRPC3 cells (TRPC3Low cells) were constructed using U-87MG cells. Thrombin (2.0 U/mL)-stimulated Ca2+ mobilization was measured by ratiometric fluorimetry. Changes in TRPC3 and S100B expression levels were determined by quantitative reverse transcription-polymerase chain reaction and immunoblotting, respectively. Cell proliferation was also measured using the WST-8 assay. RESULTS: In this cell model, thrombin-stimulated Ca2+ mobilization, and both TRPC3 and S100B expression were suppressed by chronic LiCl pretreatment and the knockdown of TRPC3. Additionally, cell proliferation was attenuated in TRPC3Low cells, compared with the negative control vector-transfected cell. CONCLUSIONS: The reduced Ca2+ mobilization and S100B expression levels following chronic LiCl pretreatment and in TRPC3Low cells support the notion that TRPC3 modulates S100B expression and is the target of the LiCl effect. Downregulation of TRPC3 may be an important mechanism by which lithium ameliorates pathophysiological intracellular Ca2+ disturbances as observed in BD, accounting, in part, for its mood-stabilizing effects.
Assuntos
Transtorno Bipolar , Sinalização do Cálcio , Cálcio/metabolismo , Lítio/farmacologia , Subunidade beta da Proteína Ligante de Cálcio S100/metabolismo , Canais de Cátion TRPC/metabolismo , Antimaníacos/farmacologia , Astrocitoma/metabolismo , Transtorno Bipolar/tratamento farmacológico , Transtorno Bipolar/metabolismo , Sinalização do Cálcio/efeitos dos fármacos , Sinalização do Cálcio/fisiologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Humanos , Trombina/farmacologiaRESUMO
OBJECTIVES: Recent findings implicate the calcium-permeable nonselective ion channels transient receptor potential (TRP) melastatin subtype 2 (TRPM2) and canonical subtype 3 (TRPC3) in the pathogenesis of bipolar disorder (BD). These channels are involved in calcium and oxidative stress signaling, both of which are disrupted in BD. Thus, we sought to determine if these channels are differentially affected by oxidative stress in cell lines of BD patient origin. METHODS: B lymphoblast cell lines (BLCLs) from bipolar I disorder (BD-I) patients (n = 6) and healthy controls (n = 5) were challenged with the oxidative stressor rotenone (2.5 µM and 10 µM) or vehicle for acute (24 hours) and chronic (four days) intervals. Cell viability was measured using propidium iodide, while TRPM2- and TRPC3-mediated calcium fluxes were measured in the presence of their respective activators (H(2) O(2) and 1-oleoyl-2-acetyl-sn-glycerol) using Fluo-4. Changes in TRPM2 and TRPC3 expression levels were determined by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and Western blotting. RESULTS: Cell viability decreased with increasing dose and duration of rotenone treatment, with BD-I patient BLCLs more susceptible than controls acutely (p < 0.001). A dose-dependent decrease in TRPC3 protein expression occurred after chronic (24%, p = 0.008) but not acute rotenone treatment. Interestingly, H(2) O(2) -provoked TRPM2-dependent calcium fluxes revealed an interaction between the effects of stressor addition and diagnostic subject group (p = 0.003). CONCLUSIONS: These data support an important role for TRPM2 and TRPC3 in sensing and responding to oxidative stress and in transducing oxidative stress signaling to intracellular calcium homeostasis and cellular stress responses, all of which have been implicated in the pathophysiology of BD.
Assuntos
Linfócitos B/metabolismo , Transtorno Bipolar/patologia , Estresse Oxidativo/fisiologia , Canais de Cátion TRPC/metabolismo , Canais de Cátion TRPM/metabolismo , Adulto , Linfócitos B/efeitos dos fármacos , Transtorno Bipolar/imunologia , Cálcio/metabolismo , Sobrevivência Celular , Relação Dose-Resposta a Droga , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Heme Oxigenase-1/genética , Heme Oxigenase-1/metabolismo , Humanos , Inseticidas/farmacologia , Masculino , Estresse Oxidativo/efeitos dos fármacos , RNA Mensageiro/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Rotenona/farmacologia , Canais de Cátion TRPC/genética , Canais de Cátion TRPM/genética , Fatores de Tempo , Adulto JovemRESUMO
The olfactory epithelium (OE) contains neural precursor cells which can be easily harvested from a minimally invasive nasal biopsy, making them a valuable cell source to study human neural cell lineages in health and disease. Glycogen synthase kinase-3 (GSK-3) has been implicated in the etiology and treatment of neuropsychiatric disorders and also in the regulation of murine neural precursor cell fate in vitro and in vivo. In this study, we examined the impact of decreased GSK-3 activity on the fate of adult human OE neural precursors in vitro. GSK-3 inhibition was achieved using ATP-competitive (6-bromoindirubin-3'-oxime and CHIR99021) or substrate-competitive (TAT-eIF2B) inhibitors to eliminate potential confounding effects on cell fate due to off-target kinase inhibition. GSK-3 inhibitors decreased the number of neural precursor cells in OE cell cultures through a reduction in proliferation. Decreased proliferation was not associated with a reduction in cell survival but was accompanied by a reduction in nestin expression and a substantial increase in the expression of the neuronal differentiation markers MAP1B and neurofilament (NF-M) after 10 days in culture. Taken together, these results suggest that GSK-3 inhibition promotes the early stages of neuronal differentiation in cultures of adult human neural precursors and provide insights into the mechanisms by which alterations in GSK-3 signaling affect adult human neurogenesis, a cellular process strongly suspected to play a role in the etiology of neuropsychiatric disorders.
Assuntos
Diferenciação Celular , Proliferação de Células , Quinase 3 da Glicogênio Sintase/antagonistas & inibidores , Células-Tronco Neurais/citologia , Neurônios/citologia , Mucosa Olfatória/citologia , Animais , Linhagem Celular , Linhagem da Célula , Sobrevivência Celular , Quinase 3 da Glicogênio Sintase/metabolismo , Humanos , Camundongos , Células-Tronco Neurais/imunologia , Células-Tronco Neurais/metabolismo , Neurogênese , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Mucosa Olfatória/enzimologia , Mucosa Olfatória/metabolismo , Nervo Olfatório/metabolismo , Transdução de SinaisRESUMO
OBJECTIVES: Disrupted intracellular calcium (Ca(2+) ) homeostasis (ICH) related to mitochondrial and/or endoplasmic reticulum (ER) dysfunction has been implicated in bipolar disorder (BD). The anti-apoptotic protein B-cell CLL/lymphoma 2 (Bcl-2), encoded in a putative BD susceptibility locus, modulates ER-Ca(2+) dynamics. Recently, an intronic single-nucleotide polymorphism (SNP) in the Bcl-2 gene, rs956572, was suggested as a functionally active SNP that influences its messenger RNA (mRNA) and protein level as well as human gray matter volume. We sought to evaluate the impact of this variant on ICH in BD. METHODS: Basal intracellular Ca(2+) concentrations ([Ca(2+) ](B) ) and rs956572 genotypes were determined in B lymphoblast cell lines (BLCLs) from bipolar I disorder (BD-I) (n=150), bipolar II disorder (BD-II) (n=65), and major depressive disorder (n=30) patients, and from healthy subjects (n=70). Bcl-2 mRNA and protein levels were determined by quantitative reverse transcriptase polymerase chain reaction and immunoblotting, respectively. Functional interactions of rs956572 with ICH were assessed by thapsigargin- and lysophosphatidic acid (LPA)-stimulated Ca(2+) responses. RESULTS: Although rs956572 variation was not significantly associated with BD, BD-I, or BD-II, BLCL [Ca(2+) ](B) was significantly higher in BD-I G/G patients compared with other genotypes and with healthy subjects. Bcl-2 mRNA and protein levels were lowest in BD-I G/G patients. Compared with A carriers, BD-I patients with G/G variants showed a modest enhancing effect on thapsigargin- and LPA-stimulated Ca(2+) responses. CONCLUSIONS: These findings support the notion that genetic variation in Bcl-2 affecting its expression impacts ICH in BD. Moreover, we show here for the first time that this interactive effect is diagnostically specific to BD-I.
Assuntos
Transtorno Bipolar/genética , Cálcio/metabolismo , Transtorno Depressivo Maior/genética , Retículo Endoplasmático/metabolismo , Genes bcl-2/genética , Homeostase , RNA Mensageiro/genética , Adulto , Linfócitos B/metabolismo , Transtorno Bipolar/metabolismo , Estudos de Casos e Controles , Transtorno Depressivo Maior/metabolismo , Retículo Endoplasmático/genética , Feminino , Genes bcl-2/fisiologia , Genótipo , Humanos , Immunoblotting , Leucemia Linfocítica Crônica de Células B/genética , Leucemia Linfocítica Crônica de Células B/metabolismo , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Adulto JovemRESUMO
Animal data indicate that the recreational drug ecstasy (3,4-methylenedioxymethamphetamine) can damage brain serotonin neurons. However, human neuroimaging measurements of serotonin transporter binding, a serotonin neuron marker, remain contradictory, especially regarding brain areas affected; and the possibility that structural brain differences might account for serotonin transporter binding changes has not been explored. We measured brain serotonin transporter binding using [(11)C] N,N-dimethyl-2-(2-amino-4-cyanophenylthio) benzylamine in 50 control subjects and in 49 chronic (mean 4 years) ecstasy users (typically one to two tablets bi-monthly) withdrawn from the drug (mean 45 days). A magnetic resonance image for positron emission tomography image co-registration and structural analyses was acquired. Hair toxicology confirmed group allocation but also indicated use of other psychoactive drugs in most users. Serotonin transporter binding in ecstasy users was significantly decreased throughout all cerebral cortices (range -19 to -46%) and hippocampus (-21%) and related to the extent of drug use (years, maximum dose), but was normal in basal ganglia and midbrain. Substantial overlap was observed between control and user values except for insular cortex, in which 51% of ecstasy user values fell below the lower limit of the control range. Voxel-based analyses confirmed a caudorostral gradient of cortical serotonin transporter binding loss with occipital cortex most severely affected. Magnetic resonance image measurement revealed no overall regional volume differences between groups; however, a slight left-hemispheric biased cortical thinning was detected in methamphetamine-using ecstasy users. The serotonin transporter binding loss was not related to structural changes or partial volume effect, use of other stimulant drugs, blood testosterone or oestradiol levels, major serotonin transporter gene promoter polymorphisms, gender, psychiatric status, or self-reported hyperthermia or tolerance. The ecstasy group, although 'grossly behaviourally normal', reported subnormal mood and demonstrated generally modest deficits on some tests of attention, executive function and memory, with the latter associated with serotonin transporter decrease. Our findings suggest that the 'typical'/low dose (one to two tablets/session) chronic ecstasy-polydrug user might display a highly selective mild to marked loss of serotonin transporter in cerebral cortex/hippocampus in the range of that observed in Parkinson's disease, which is not gender-specific or completely accounted for by structural brain changes, recent use of other drugs (as assessed by hair analyses) or other potential confounds that we could address. The striking sparing of serotonin transporter-rich striatum (although possibly affected in 'heavier' users) suggests that serotonergic neurons innervating cerebral cortex are more susceptible, for unknown reasons, to ecstasy than those innervating subcortical regions and that behavioural problems in some ecstasy users during abstinence might be related to serotonin transporter changes limited to cortical regions.
Assuntos
Transtornos Relacionados ao Uso de Anfetaminas/metabolismo , Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/metabolismo , Alucinógenos/farmacologia , N-Metil-3,4-Metilenodioxianfetamina/farmacologia , Proteínas da Membrana Plasmática de Transporte de Serotonina/metabolismo , Adulto , Transtornos Relacionados ao Uso de Anfetaminas/diagnóstico por imagem , Transtornos Relacionados ao Uso de Anfetaminas/patologia , Benzilaminas/metabolismo , Encéfalo/diagnóstico por imagem , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Radioisótopos de Carbono , Córtex Cerebral/diagnóstico por imagem , Doença Crônica , Feminino , Hormônios/sangue , Humanos , Imageamento por Ressonância Magnética , Masculino , Testes Neuropsicológicos , Tomografia por Emissão de Pósitrons , Escalas de Graduação Psiquiátrica , Proteínas da Membrana Plasmática de Transporte de Serotonina/genética , Sono , Inquéritos e QuestionáriosRESUMO
Suicide and suicidal behaviour are a major health concern worldwide particularly in patients with mood disorders. Family, adoption and twin studies show that genetics influences suicidal behaviour. The serotonin transporter (5HTT) plays an important role in the pathophysiology of mood disorders and may also be involved in suicidal behaviour since 5HTT binding is decreased in the brain of suicide completers. Because the effect of genomic imprinting in the 5HTT gene on suicidal behaviour has not been investigated, we analysed the parent-of-origin effect (POE) of four 5HTT markers and the differential expression of the 5HTT G2651T (rs1042173) alleles in suicide attempters affected by bipolar disorder. We performed a family based association study and ETDT/QTDT analyses of the rs25531, HTTLPR, VNTR-2 and G2651T polymorphisms in 312 nuclear families with at least one subject affected by bipolar disorder. The main outcomes investigated in this study are bipolar disorder diagnosis, suicide attempts, suicidal behaviour severity and age at onset of bipolar disorder. We also compared the allele-specific mRNA levels in lymphoblastoid cells from 13 bipolar suicide attempters and 8 bipolar non-suicide attempters. Allele 2651T was transmitted significantly more often to bipolar patients (P = 0.042). There was no significant difference between maternal and paternal transmission ratios. Furthermore, there was no significant difference in the ratio of T/G-specific mRNA expression between bipolar attempters and non-attempters. These data do not support a role for differential allelic expression of 5HTT for suicidal behaviour in bipolar disorder. Small sample size and the fact that RNA was obtained from lymphoblastoid cell lines were some of the limitations of this study.
Assuntos
Transtorno Bipolar/genética , Transtorno Bipolar/metabolismo , Proteínas da Membrana Plasmática de Transporte de Serotonina/genética , Ideação Suicida , Tentativa de Suicídio/psicologia , Regiões 3' não Traduzidas/genética , Adulto , Idade de Início , Alcoolismo/complicações , Alcoolismo/psicologia , Alelos , DNA/genética , Manual Diagnóstico e Estatístico de Transtornos Mentais , Feminino , Expressão Gênica/fisiologia , Estudos de Associação Genética , Humanos , Íntrons/genética , Masculino , Pessoa de Meia-Idade , Pais , Polimorfismo Genético/genética , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Transtornos Relacionados ao Uso de Substâncias/complicações , Transtornos Relacionados ao Uso de Substâncias/psicologia , Resultado do TratamentoRESUMO
Irregularities of intracellular calcium (Ca2+) homeostasis have been implicated in the pathophysiology of bipolar disorder (BD). Findings that chronic ex-vivo treatment with lithium modifies lysophosphatidic acid (LPA)-stimulated Ca2+ responses in B lymphoblast cell lines (BLCLs) from BD-I patients and healthy controls, and differentially decreases levels of the type-3 canonical transient receptor potential Ca2+-permeable channel in BLCLs from BD-I patients, support the view that the amelioration of these abnormalities is important in the therapeutic action of lithium. To determine whether other clinically efficacious mood stabilizers share these effects, LPA (100 mum)- and thapsigargin (TG, 200 nm)-stimulated Ca2+ responses were determined in BLCLs from BD-I patients and healthy controls treated acutely (24 h) and chronically (7 d) ex vivo with therapeutically relevant concentrations of lithium (0.75 mm), valproate (0.5 mm), lamotrigine (15 mum) or respective vehicles. Chronic treatment with valproate significantly attenuated LPA-stimulated Ca2+ responses ([downward arrow]8%: F's=9.1-9.4, d.f.=1, 9, p's<0.05) compared to vehicle in BLCLs from BD-I patients and healthy controls, similar to chronic lithium treatment ([downward arrow]8%: F=6.2, d.f.=1, 21, p<0.05), but also attenuated TG-evoked Ca2+ responses ([downward arrow]10% to [downward arrow]19%: F's=5.5-15.5, d.f.=1, 12, p's<0.05). However, chronic lamotrigine treatment did not affect LPA- or TG-stimulated Ca2+ responses. These results suggest that chronic lithium and valproate treatments act differently from lamotrigine in respect of modulation of receptor- and/or capacitance-mediated Ca2+ flux. These differential effects on Ca2+ responses may be relevant to the distinctive clinical profiles of these mood stabilizers.
Assuntos
Antimaníacos/farmacologia , Linfócitos B/efeitos dos fármacos , Cálcio/metabolismo , Líquido Intracelular/efeitos dos fármacos , Cloreto de Lítio/farmacologia , Adulto , Linfócitos B/metabolismo , Linfócitos B/patologia , Transtorno Bipolar/patologia , Linhagem Celular Transformada , Interações Medicamentosas , Inibidores Enzimáticos/farmacologia , Feminino , Humanos , Líquido Intracelular/metabolismo , Lipopolissacarídeos/farmacologia , Masculino , Pessoa de Meia-Idade , Tapsigargina/farmacologia , Ácido Valproico/farmacologiaRESUMO
OBJECTIVE: Recent case-control studies implicate the transient receptor potential melastatin 2 (TRPM2) channel in conferring risk for bipolar disorder (BD), though the risk variants differed. As confounding effects of population structure could not be unequivocally ruled out as the basis for the discordance, we tested the association of TRPM2 with BD in a family design, which is immune to population stratification, for those TRPM2 single nucleotide polymorphisms (SNPs) previously reported as associated with BD. METHODS: The exon 11 SNP (rs1556314) and four informative intronic SNPs (rs1785437, rs1618355, rs933151, and rs749909) were genotyped in 300 BD families by TaqMan allelic discrimination and results were analyzed using chi(2) test, transmission disequilibrium test, and pedigree-based association. SNP rs1556314 was also genotyped in our case-control sample set comprised of 184 BD and 195 healthy Caucasian subjects. RESULTS: The SNP rs1556314 in exon 11 was significantly associated with bipolar disorder type I (BD-I) (p = 0.011, p(permutation) = 0.015) in the case-control dataset and in the family design (p = 0.018, p(permutation) = 0.052, TDTPHASE). Interestingly, the C-T-A haplotype of SNPs rs1618355, rs933151, and rs749909 was significantly associated with early age at onset in BD-I families. CONCLUSION: Significant association of TRPM2 genetic variants with BD in case-control and family datasets further supports a role for TRPM2 in the pathogenesis of this disorder. Overtransmission of the G allele of rs1556314 at exon 11 of TRPM2 in BD-I but not bipolar disorder type II (BD-II) further supports different genetic contributions to the pathogenesis of these bipolar phenotypes.
Assuntos
Transtorno Bipolar/genética , Saúde da Família , Predisposição Genética para Doença , Polimorfismo de Nucleotídeo Único/genética , Canais de Cátion TRPM/genética , Adolescente , Adulto , Idade de Início , Idoso , Transtorno Bipolar/classificação , Transtorno Bipolar/etiologia , Estudos de Casos e Controles , Distribuição de Qui-Quadrado , Éxons/genética , Feminino , Frequência do Gene , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Risco , Adulto JovemRESUMO
This study investigated whether abnormalities in serotonin transporter binding occur in Parkinson's disease (PD) patients with concurrent depression. We estimated serotonin transporter levels in seven clinically depressed early-stage PD patients and in seven healthy matched-control subjects during a single positron emission tomography (PET) scan with the serotonin transporter radioligand, [(11)C]DASB. Depressed PD patients displayed a wide-spread increase (8-68%) in [(11)C]DASB specific binding outside of the striatum, which was significant in dorsolateral (37%) and prefrontal (68%) cortices. Elevated [(11)C]DASB binding was positively correlated with depressive symptoms but not with disease severity or duration. Compatible with recent PET/[(11)C]DASB findings in major depression, the present preliminary data suggest that increased [(11)C]DASB binding, possibly reflecting greater serotonin transporter density (up-regulation), might be a pathological feature of depression in Parkinson's disease-and possibly a characteristic of depressive illness in general.
Assuntos
Compostos de Anilina , Depressão/diagnóstico por imagem , Depressão/etiologia , Doença de Parkinson/complicações , Tomografia por Emissão de Pósitrons/métodos , Proteínas da Membrana Plasmática de Transporte de Serotonina/metabolismo , Sulfetos , Idoso , Estudos de Casos e Controles , Depressão/patologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Doença de Parkinson/diagnóstico por imagem , Córtex Pré-Frontal/diagnóstico por imagem , Ligação Proteica/efeitos dos fármacos , Escalas de Graduação Psiquiátrica , Estatísticas não ParamétricasRESUMO
We have previously reported an association between the BDNF Val66Met polymorphism and bipolar disorder (BD). However, the possibility that genomic imprinting in BDNF gene affects risk for BD has not been investigated. To examine the possibility of genomic imprinting in the BDNF gene in BD, we analyzed the parent-of-origin effect (POE) and differential expression of the BDNF Val66Met alleles in BD. We performed a family-based association study and ETDT analyses of the Val66Met polymorphism in 312 BD nuclear families, and compared allele-specific mRNA levels in both post-mortem brain samples and B lymphoblasts from BD patients and controls. The BDNF Val66 allele was transmitted significantly more often to patients with BD (maternal transmissions: 46/22, p=0.003; paternal transmissions: 55/30, p=0.006). There was no significant difference between maternal and paternal transmission ratios. There was no significant difference in the ratio of Val/Met-specific mRNA expression between BD and controls, in either brain or B lymphoblasts. The Val/Met ratio was much lower in the brain vs. B lymphoblasts. These data do not support a role for genomic imprinting as a modifier of the contribution of BDNF gene to risk of susceptibility to BD.
Assuntos
Transtorno Bipolar/genética , Fator Neurotrófico Derivado do Encéfalo/genética , Metionina/genética , Polimorfismo Genético/genética , RNA Mensageiro/metabolismo , Valina/genética , Adulto , Alelos , Linfócitos B/metabolismo , Transtorno Bipolar/patologia , Encéfalo/metabolismo , Distribuição de Qui-Quadrado , Saúde da Família , Feminino , Frequência do Gene , Humanos , Masculino , Pessoa de Meia-Idade , Mudanças Depois da MorteRESUMO
BACKGROUND: Aberrant intracellular calcium (Ca2+) signaling in patients with bipolar-I disorder (BD-I) suggests disturbed endoplasmic reticulum (ER) function in BD. We examined whether the ER stress response is altered in BD-I patients and the relationship to basal intracellular Ca2+ levels ([Ca2+]B), in B lymphoblasts (BLCLs) from BD-I patients. METHODS: Endoplasmic reticulum stress-induced X-box binding protein 1 (XBP1), C/EBP homologous protein (CHOP), and GRP78 expression in BLCLs from BD-I subjects stratified on elevated or normal [Ca2+]B and control subjects were determined by real-time quantitative reverse transcription polymerase chain reaction. The XBP1 -116C/G polymorphism, which impairs the XBP1 loop in the ER stress response, were genotyped with a TaqMan-based assay. RESULTS: Compared with control subjects, thapsigargin- and tunicamycin-induced increases in XBP1 and CHOP but not GRP78 messenger RNA levels were significantly lower in BD-I patients. However, induction of these genes did not differ significantly in the two BD-I subgroups stratified on [Ca2+]B. Furthermore, the attenuated XBP1 induction cannot be explained solely by differences of XBP1 -116C/G genotype frequency. CONCLUSIONS: Our findings suggest that the ER stress response is impaired in BD-I patients but irrespective of altered intracellular Ca2+ homeostasis as reflected in elevated [Ca2+]B. Moreover, an effect of XBP1 -116C/G polymorphism could not account for the attenuated XBP1 induction in bipolar-I disorder observed in this study.
Assuntos
Linfócitos B/fisiologia , Transtorno Bipolar/fisiopatologia , Sinalização do Cálcio/genética , Cálcio/fisiologia , Proteínas de Ligação a DNA/genética , Retículo Endoplasmático/fisiologia , Proteínas de Choque Térmico/genética , Chaperonas Moleculares/genética , Proteínas Nucleares/genética , Adulto , Linfócitos B/metabolismo , Transtorno Bipolar/genética , Transtorno Bipolar/metabolismo , Cálcio/metabolismo , Sinalização do Cálcio/fisiologia , Linhagem Celular , Proteínas de Ligação a DNA/metabolismo , Proteínas de Ligação a DNA/fisiologia , Retículo Endoplasmático/genética , Retículo Endoplasmático/metabolismo , Chaperona BiP do Retículo Endoplasmático , Feminino , Proteínas de Choque Térmico/metabolismo , Humanos , Masculino , Chaperonas Moleculares/metabolismo , Proteínas Nucleares/metabolismo , Proteínas Nucleares/fisiologia , Polimorfismo de Nucleotídeo Único , RNA Mensageiro/metabolismo , Fatores de Transcrição de Fator Regulador X , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Transcrição CHOP/genética , Fator de Transcrição CHOP/metabolismo , Fatores de Transcrição , Proteína 1 de Ligação a X-BoxRESUMO
Despite much evidence that lithium and valproate, two commonly used mood stabilizers, exhibit neuroprotective properties against an array of insults, the pharmacological relevance of such effects is not clear because most of these studies examined the acute effect of these drugs in supratherapeutic doses against insults which were of limited disease relevance to bipolar disorder. In the present study, we investigated whether lithium and valproate, at clinically relevant doses, protects human neuroblastoma (SH-SY5Y) and glioma (SVG and U87) cells against oxidative stress and endoplasmic reticulum stress in a time-dependent manner. Pretreatment of SH-SY5Y cells for 7 days, but not 1 day, with 1 mM of lithium or 0.6 mM of valproate significantly reduced rotenone and H2O2-induced cytotoxicity, cytochrome c release and caspase-3 activation, and increased Bcl-2 levels. Conversely, neither acute nor chronic treatment of SH-SY5Y cells with lithium or valproate elicited cytoprotective responses against thapsigargin-evoked cell death and caspase-3 activation. Moreover, inhibitors of glycogen synthase kinase-3 (GSK-3), kenpaullone and SB216763, abrogated rotenone-induced, but not H2O2-induced, cytotoxicity. Thus the cytoprotective effects of lithium and valproate against H2O2-induced cell death is likely independent of GSK-3 inhibition. On the other hand, chronic lithium or valproate treatment did not ameliorate cytotoxicity induced by rotenone, H2O2, and thapsigargin in SVG astroglial and U87 MG glioma cell lines. Our results suggest that lithium and valproate may decrease vulnerability of human neural, but not glial, cells to cellular injury evoked by oxidative stress possibly arising from putative mitochondrial disturbances implicated in bipolar disorder.
Assuntos
Antimaníacos/farmacologia , Retículo Endoplasmático/fisiologia , Compostos de Lítio/farmacologia , Fármacos Neuroprotetores/farmacologia , Estresse Oxidativo , Ácido Valproico/farmacologia , Caspase 3 , Caspases/metabolismo , Morte Celular , Linhagem Celular Tumoral , Citocromos c/metabolismo , Citoproteção , Complexo I de Transporte de Elétrons/antagonistas & inibidores , Retículo Endoplasmático/efeitos dos fármacos , Ativação Enzimática , Inibidores Enzimáticos/farmacologia , Glioma , Quinase 3 da Glicogênio Sintase/antagonistas & inibidores , Glicogênio Sintase Quinase 3 beta , Humanos , Peróxido de Hidrogênio/farmacologia , Neuroblastoma , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Rotenona/farmacologia , Tapsigargina/farmacologia , Regulação para CimaRESUMO
Altered 1-oleoyl-lysophosphatidic acid (LPA, 100 microM)-stimulated calcium responses occur in B-lymphoblast cell lines from bipolar disorder patients, but the mechanism(s) involved is uncertain. Lysophosphatidic acid shares a structurally similar fatty acid side chain with the diacylglycerol analogue, 1-oleoyl-2-acetyl-sn-glycerol (OAG), a known activator of subtypes 3, 6 and 7 of the canonical transient receptor potential (TRPC) cation channel subfamily. Accordingly, the objective of this study was to determine whether the LPA-stimulated calcium response in B-lymphoblasts is mediated, in part, through this TRPC channel subfamily. Divalent cation selectivity in response to thapsigargin, LPA and OAG were used to distinguish TRPC-like character of the responses to these agents in BLCLs. The sensitivity to gadolinium, an inhibitor of capacitative calcium channels, was used to determine the store-operated nature of the responses. The TRPC isoforms that are present in BLCLs as identified by immunoblotting and/or PCR include TRPC1, 3 and 5. Minimal barium influx in calcium-free buffer was observed following thapsigargin stimulation. However, LPA stimulated barium influx of a magnitude similar to that induced by OAG. Thapsigargin-provoked calcium influx was completely inhibited by gadolinium (10 microM), whereas LPA and OAG-stimulated responses were partially inhibited and potentiated, respectively. The results suggest that 100 microM LPA stimulates calcium entry through channels with characteristics similar to TRPC3, as TRPC6 and 7 are absent in B-lymphoblasts.
Assuntos
Linfócitos B , Transtorno Bipolar/metabolismo , Sinalização do Cálcio/efeitos dos fármacos , Lisofosfolipídeos/farmacologia , Canais de Cátion TRPC/metabolismo , Linfócitos B/efeitos dos fármacos , Linfócitos B/metabolismo , Bário/metabolismo , Linhagem Celular , Diglicerídeos/farmacologia , Humanos , Immunoblotting , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais/efeitos dos fármacos , Tapsigargina/farmacologiaRESUMO
OBJECTIVES: Intracellular calcium (Ca(2+)) dyshomeostasis (ICDH) has been implicated in bipolar disorder (BD) pathophysiology. We previously showed that SNP rs956572 in the B-cell CLL/lymphoma 2 (Bcl-2) gene associates with elevated B lymphoblast (BLCL) intracellular Ca(2+) concentrations ([Ca(2+)]B) differentially in BD-I. Genome-wide association studies strongly support the association between BD and the SNP rs1006737, located within the L-type voltage-dependent Ca(2+) channel α1C subunit gene (CACNA1C). Here we investigated whether this CACNA1C variant also associates with ICDH and interacts with SNP rs956572 on [Ca(2+)]B in BD-I. METHODS: CACNA1C SNP rs1006737 was genotyped in 150 BD-I, 65 BD-II, 30 major depressive disorder patients, and 70 healthy subjects with available BLCL [Ca(2+)]B and Bcl-2 SNP rs956572 genotype measures. RESULTS: SNP rs1006737 was significantly associated with BD-I. The [Ca(2+)]B was significantly higher in BD-I rs1006737 A compared with healthy A allele carriers and also in healthy GG compared with A allele carriers. There was no significant interaction between SNP rs1006737 and SNP rs956572 on [Ca(2+)]B. CONCLUSIONS: Our study further supports the association of SNP rs1006737 with BD-I and suggests that CACNA1C SNP rs1006737 and Bcl-2 SNP rs956572, or specific causal variants in LD with these proxies, act independently to increase risk and ICDH in BD-I.
Assuntos
Transtorno Bipolar/genética , Canais de Cálcio Tipo L/genética , Cálcio/metabolismo , Homeostase , Polimorfismo de Nucleotídeo Único , Proteínas Proto-Oncogênicas c-bcl-2/genética , Adolescente , Adulto , Alelos , Canadá , Estudos de Casos e Controles , Feminino , Predisposição Genética para Doença , Estudo de Associação Genômica Ampla , Heterozigoto , Humanos , Masculino , Adulto JovemRESUMO
Cardiovascular and hypothalamic pituitary axis (HPA) disturbances have been observed in individuals who are pathological gamblers (PGs). These may partly derive from chronic exposure to gambling. Response to amphetamine (AMPH) may reveal such disturbances while controlling for differential conditioned responses to gambling in PGs vs healthy controls (HCs). This study assessed heart rate (HR), systolic blood pressure (SBP) and diastolic blood pressure (DBP) and plasma cortisol following oral AMPH (0.4 mg/kg) in male PGs (n=12) and HCs (n=11) who underwent a positron emission tomography (PET) scan. The Stop Signal Task enabled assessment of the link between physiological and behavioral dysregulation. Trait moderating effects were explored. The responses of PGs to AMPH differed from those of HCs on every index. PGs displayed persistent elevation in DBP and concomitant reduction in HR (i.e. baroreflex) compared to HCs beyond 90 min post-dose. PGs displayed deficits in cortisol compared to HCs that were partially reversed by AMPH. Impairment on the Stop Signal Task correlated positively with HR in controls, but negatively with HR in PGs, suggesting that strong initial and compensatory cardiac responses to a stimulant may each predict disinhibition. Extraversion predicted greater disinhibition in PGs. Noradrenergic disturbances may contribute to sensitized responses to stimulant challenge and disinhibition in PGs.
Assuntos
Anfetamina/efeitos adversos , Sistema Cardiovascular/efeitos dos fármacos , Estimulantes do Sistema Nervoso Central/efeitos adversos , Jogo de Azar/induzido quimicamente , Hipotálamo/efeitos dos fármacos , Hipófise/efeitos dos fármacos , Adulto , Pressão Sanguínea/efeitos dos fármacos , Estudos de Casos e Controles , Jogo de Azar/sangue , Frequência Cardíaca/efeitos dos fármacos , Humanos , Hidrocortisona/sangue , Hipotálamo/metabolismo , Masculino , Tomografia por Emissão de Pósitrons/métodosRESUMO
Plasma-free tryptophan is inversely related to the severity of subjective pain in 8 patients who fulfilled criteria for a variety of non-articular rheumatism, the "fibrositis syndrome". The observation is consistent with animal and human studies suggesting a relationship between reduced brain serotonin metabolism and pain reactivity.
Assuntos
Fibromialgia/fisiopatologia , Dor/etiologia , Triptofano/sangue , Adulto , Feminino , Fibromialgia/sangue , Humanos , Masculino , Doenças Reumáticas , Serotonina/fisiologia , SíndromeRESUMO
Elevated basal intracellular calcium (Ca(2+)) levels ([Ca(2+)](B)) in B lymphoblast cell lines (BLCLs) from bipolar I disorder (BD-I) patients implicate altered Ca(2+) homeostasis in this illness. Chronic lithium treatment affects key proteins modulating intracellular Ca(2+) signaling. Thus, we sought to determine if chronic exposure to therapeutic lithium concentrations also modifies intracellular Ca(2+) homeostasis in this surrogate cellular model of signal transduction disturbances in BD. BLCLs from BD-I (N=26) and healthy subjects (N=17) were regrown from frozen stock and incubated with 0.75 mM lithium or vehicle for 24 h (acute) or 7 days (chronic). [Ca(2+)](B), lysophosphatidic acid (LPA)-stimulated Ca(2+) mobilization ([Ca(2+)](S)), and thapsigargin-induced store-operated Ca(2+) entry (SOCE) were determined using ratiometric fluorometry with Fura-2. Compared with vehicle, chronic lithium exposure resulted in significantly higher [Ca(2+)](B) (F=8.47; p=0.006) in BLCLs from BD-I and healthy subjects. However, peak LPA-stimulated [Ca(2+)](S) and SOCE were significantly reduced (F=11.1, p=0.002 and F=8.36, p=0.007, respectively). Acute lithium exposure did not significantly affect measured parameters. In summary, the effect of chronic lithium to elevate [Ca(2+)](B) in BLCLs while attenuating both receptor-stimulated and SOCE components of intracellular Ca(2+) mobilization in BLCLs suggests that modulation of intracellular Ca(2+) homeostasis may be important to the therapeutic action of lithium.
Assuntos
Cálcio/metabolismo , Homeostase/efeitos dos fármacos , Espaço Intracelular/efeitos dos fármacos , Lítio/farmacologia , Adulto , Linfócitos B/efeitos dos fármacos , Linfócitos B/virologia , Transtorno Bipolar/tratamento farmacológico , Estudos de Casos e Controles , Contagem de Células , Linhagem Celular , Transformação Celular Viral/efeitos dos fármacos , Distribuição de Qui-Quadrado , Esquema de Medicação , Inibidores Enzimáticos/farmacologia , Feminino , Fura-2/metabolismo , Herpesvirus Humano 4/metabolismo , Humanos , Espaço Intracelular/metabolismo , Lítio/uso terapêutico , Lisofosfolipídeos/farmacologia , Masculino , Análise Multivariada , Tapsigargina/farmacologia , Fatores de TempoRESUMO
Earlier findings of elevated basal and stimulated PKA activities, and increased immunoreactive levels of PKA regulatory and catalytic subunits in discrete postmortem brain regions from bipolar disorder (BD) patients suggest that disturbances in PKA are involved in the pathophysiology of BD. PKA subunit mRNA levels were measured using SYBR Green real-time RT-PCR to determine if previously observed differences in immunoreactive levels of PKA RIIbeta and Calpha subunits were associated with corresponding changes in mRNA levels in temporal and frontal cortices from the same BD patients and matched controls. In distinct contrast to the higher immunolabeling levels of the PKA subunits previously reported in the BD brain, there were no significant differences in RIIbeta and Calpha subunit mRNA levels in the temporal and frontal cortices of BD patients compared with controls. These findings infer that the elevated PKA immunolabeling and activity found in the selected cerebral cortical regions of BD postmortem brain were due to a posttranscriptional mechanism, rather than changes in regulation of gene transcription and/or mRNA stability of the PKA subunits.
Assuntos
Transtorno Bipolar/metabolismo , Encéfalo/metabolismo , Domínio Catalítico/genética , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Northern Blotting , Estudos de Casos e Controles , Proteínas Quinases Dependentes de AMP Cíclico/genética , Primers do DNA/metabolismo , DNA Complementar/química , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Mudanças Depois da Morte , Isoformas de Proteínas/genética , RNA/análise , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Estatística como AssuntoRESUMO
Recent studies in rat brain and cell cultures have demonstrated that expression of the peptide-folding chaperone protein calreticulin is increased by valproate treatment, while the anti-apoptotic Bcl-2 is increased by both lithium and valproate. We asked whether a similar pattern of regulation by these drugs is evident in human neuronal and glial cells. One-week treatment with 1 mM valproate induced a significant (90%) increase in calreticulin mRNA and protein levels in SVG, a glial cell line, but reduced its mRNA levels by 38% in hNT neuronal cells. Valproate also markedly increased Bcl-2 mRNA levels by 260%, but only in hNT neurons. In contrast, lithium had no significant effect in either cell type. Valproate-induced increases in calreticulin may therefore show glial specificity in humans, while changes in Bcl-2 levels may be neuron specific. These results highlight the cell model dependence of outcomes in molecular studies of mood stabilizer effects and the need for caution in interpreting findings in model systems.