RESUMO
Cadmium (Cd) is a heavy metal that is one of the most toxic environmental pollutants throughout the world. We previously reported that Cd exposure impairs olfactory memory in mice. However, the underlying mechanisms for its neurotoxicity for olfactory function are not well understood. Since adult Subventricular zone (SVZ) and Olfactory Bulb (OB) neurogenesis contributes to olfaction, olfactory memory defects caused by Cd may be due to inhibition of neurogenesis. In this study, using bromodeoxyuridine (BrdU) labeling and immunohistochemistry, we found that 0.6 mg/L Cd exposure through drinking water impaired adult SVZ/OB neurogenesis in C57BL/6 mice. To determine if the inhibition of olfactory memory by Cd can be reversed by stimulating adult neurogenesis, we utilized the transgenic caMEK5 mouse strain to conditional stimulate of adult neurogenesis by activating the endogenous ERK5 MAP kinase signaling pathway. This was accomplished by conditionally induced expression of active MEK5 (caMEK5) in adult neural stem/progenitor cells. The caMEK5 mice were exposed to 0.6 mg/L Cd for 38 weeks, and tamoxifen was administered to induce caMEK5 expression and stimulate adult SVZ/OB neurogenesis during Cd exposure. Short-term olfactory memory test and sand-digging based, odor-cued olfactory learning and memory test were conducted after Cd and tamoxifen treatments to examine their effects on olfaction. Here we report that Cd exposure impaired short-term olfactory memory and odor-cued associative learning and memory in mice. Furthermore, the Cd-impaired olfactory memory deficits were rescued by the tamoxifen-induction of caMEK5 expression. This suggests that Cd exposure impairs olfactory function by affecting adult SVZ/OB neurogenesis in mice.
Assuntos
Comportamento Animal , Ventrículos Laterais/enzimologia , Memória , Proteína Quinase 7 Ativada por Mitógeno/metabolismo , Neurogênese , Transtornos do Olfato/prevenção & controle , Bulbo Olfatório/enzimologia , Percepção Olfatória , Olfato , Animais , Aprendizagem por Associação , Cloreto de Cádmio , Sinais (Psicologia) , Modelos Animais de Doenças , Ativação Enzimática , Feminino , Ventrículos Laterais/patologia , Ventrículos Laterais/fisiopatologia , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Proteína Quinase 7 Ativada por Mitógeno/genética , Odorantes , Transtornos do Olfato/induzido quimicamente , Transtornos do Olfato/enzimologia , Transtornos do Olfato/fisiopatologia , Bulbo Olfatório/patologia , Bulbo Olfatório/fisiopatologia , Fatores de TempoRESUMO
Olfactory dysfunction is observed in several neurological disorders, including Huntington disease (HD), and correlates with global cognitive performance, depression and degeneration of olfactory regions in the brain. Despite clear evidence demonstrating olfactory dysfunction in HD patients, only limited details are available in murine models and the underlying mechanisms are unknown. In order to determine if alterations in the olfactory bulb (OB) are observed in HD we assessed OB weight or area from 3 to 12â¯months of age in the BACHD transgenic lines (TG5 and TG9). A significant decrease in the OB was observed at 6 and 12â¯months of age compared to WT. We also detected increased mRNA and protein expression of mutant huntingtin (mHTT) in the OB of TG5 compared to TG9 at specific ages. Despite the higher expression of mHTT in the TG5 OBs, there was increased nuclear accumulation of mHTT in the OB of TG9 compared to WT and TG5 rats. As we observed atrophy of the OB in the BACHD rats we assessed for caspase activation, a known mechanism underlying the cell death observed in HD. We characterized caspase-3, -6, -8 andâ¯-â¯9 mRNA and protein expression levels in the OB of the BACHD transgenic lines at 3, 6 and 12â¯months of age. Alterations in caspase mRNA and protein expression were detected in the TG5 and TG9 lines. However, the changes observed in the mRNA and protein levels are in some cases discordant, suggesting that the caspase protein modifications detected may be more attributable to post-translational modifications. The caspase activation studies support that cell death may be increased in the rodent HD OB and further our understanding of the olfactory dysfunction and the role of caspases in the pathogenesis of HD.
Assuntos
Caspases/metabolismo , Doença de Huntington/complicações , Transtornos do Olfato/etiologia , Bulbo Olfatório/enzimologia , Bulbo Olfatório/patologia , Animais , Atrofia/etiologia , Atrofia/patologia , Modelos Animais de Doenças , Ativação Enzimática/fisiologia , Humanos , Proteína Huntingtina/genética , Doença de Huntington/enzimologia , Doença de Huntington/patologia , Transtornos do Olfato/enzimologia , Transtornos do Olfato/patologia , Ratos , Ratos TransgênicosRESUMO
Neuroinflammation has been shown to constitute a crucial mechanism in the pathophysiology of epileptic brain and several genes of inflammatory mediators have been detected in surgically resected hippocampus tissue but not in non-related seizure brain regions. Interestingly, it has been reported an olfactory dysfunction in frontal lobe epilepsy (FLE). Our aim was to quantify the gene expression of inflammatory-related and nitric oxide synthase genes in olfactory bulbs (OB) tissue from FLE patients. RNA was isolated from OB resection of FLE patients and autopsy subjects without any neurological disease (n = 7, each). After cDNA synthesis, we performed qPCR for interleukin-1ß (IL-1ß), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), nuclear factor κB p65 (RELA), Toll-like receptor 4 (TLR 4), its agonist high mobility group box 1 (HMGB 1) as well nitric oxide synthase isozymes (NOS 1, 2 and 3). We found a significant increase in gene expression of pro-inflammatory cytokines (IL-1ß, IL-6 and TNFα), TLR4 receptor and in its agonist HMGB1 and the downstream transcription factor NFκB p65. Moreover, we observed an increase of both NOS1 and NOS3 and a slightly increase of NOS2; however, it was not significant. Our study describes the overexpression of inflammatory-related genes and NOS isozymes in OB from FLE patients. Even though, the number of patients was limited, our findings could point out that neuroinflammation and nitrosative stress-related genes in the OB could be produced in general manner in all brain regions and thus contribute in part, to the olfactory dysfunction observed in FLE patients.
Assuntos
Citocinas/metabolismo , Epilepsia do Lobo Frontal/enzimologia , Epilepsia do Lobo Frontal/imunologia , Óxido Nítrico Sintase/metabolismo , Bulbo Olfatório/enzimologia , Bulbo Olfatório/imunologia , Adulto , Idoso , Criança , Epilepsia Resistente a Medicamentos/diagnóstico por imagem , Epilepsia Resistente a Medicamentos/enzimologia , Epilepsia Resistente a Medicamentos/imunologia , Epilepsia Resistente a Medicamentos/cirurgia , Epilepsia do Lobo Frontal/diagnóstico por imagem , Epilepsia do Lobo Frontal/cirurgia , Feminino , Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
Long-term fear memory formation in the hippocampus and neocortex depends upon brain-derived neurotrophic factor (BDNF) signaling after acquisition. Incremental, appetitive odor discrimination learning is thought to depend substantially on the differentiation of adult-born neurons within the olfactory bulb (OB)-a process that is closely associated with BDNF signaling. We sought to elucidate the role of neurotrophin signaling within the OB on odor memory consolidation. Male mice were trained on odor-reward associative discriminations after bilateral infusion of the kinase inhibitor K252a, or vehicle control, into the OB. K252a is a partially selective inhibitor of tyrosine kinase (Trk) receptors, including the TrkB receptor for BDNF, though it also inhibits other plasticity-related kinases such as PKC and CaMKII/IV. K252a infusion into the OB did not impair odor acquisition or short-term (2 h) memory for the learned discriminations, but significantly impaired long-term (48 h) odor memory (LTM). This LTM deficit also was associated with reduced selectivity for the conditioned odorant in a reward-seeking digging task. Infusions of K252a immediately prior to testing did not impair LTM recall. These results indicate that kinase activation in the OB is required for the consolidation of odor memory of incrementally acquired information.
Assuntos
Consolidação da Memória/fisiologia , Memória de Longo Prazo/fisiologia , Bulbo Olfatório/enzimologia , Receptores Proteína Tirosina Quinases/fisiologia , Olfato/fisiologia , Animais , Carbazóis/administração & dosagem , Discriminação Psicológica , Inibidores Enzimáticos/administração & dosagem , Alcaloides Indólicos/administração & dosagem , Masculino , Consolidação da Memória/efeitos dos fármacos , Memória de Longo Prazo/efeitos dos fármacos , Camundongos , Odorantes , Percepção Olfatória/fisiologia , Receptores Proteína Tirosina Quinases/antagonistas & inibidoresRESUMO
Arginases are enzymes of the urea cycle that catalyze the hydrolysis of l-arginine to ornithine and urea. The enzymes are core components of the arginine-ornithine-glutamate-γ-amino butyric acid pathway of the central nervous system. In the present study, we immunohistochemically determined the localization of arginase 1 and 2 in the olfactory bulb (OB) of the roe dear (Capreolus pygargus). Reverse transcription PCR revealed that the mRNAs encoding both arginase 1 and 2 were expressed in the OB. Arginase 1 was localized to olfactory nerve axons, calcitonin gene-related peptide-positive mitral/tufted cells (excitatory neurons), and glutamate acid decarboxylase 65/67-immunopositive periglomerular cells of the main olfactory bulb. The arginase 2 immunoreactivities in the OB tissues were similar to those of arginase 1. Furthermore, both arginases were detected in the accessory olfactory bulb. These findings suggest that both arginase 1 and 2 are potentially associated with excitatory and inhibitory neurotransmitter activities in animal OBs, including those of the roe deer.
Assuntos
Arginase/metabolismo , Cervos/fisiologia , Bulbo Olfatório/enzimologia , Transmissão Sináptica/fisiologia , Animais , Arginase/genética , Cervos/genética , Imuno-Histoquímica , República da Coreia , Transmissão Sináptica/genéticaRESUMO
Trehalose is a nonreducing disaccharide that has recently attracted much attention because of its ability to inhibit protein aggregation, induce autophagy, and protect against dissections and strokes. In vertebrates, the biosynthesis of trehalose was long considered absent due to the lack of annotated genes involved in this process. In contrast, trehalase (TreH), which is an enzyme required for the cleavage of trehalose, is known to be conserved and expressed. Here, we show that trehalose is present as an endogenous metabolite in the rodent hippocampus. We found that primary astrocytes were able to synthesize trehalose and release it into the extracellular space. Notably, the TreH enzyme was observed only in the soma of neurons, which are the exclusive users of this substrate. A statistical analysis of the metabolome during different stages of maturation indicated that this metabolite is implicated in neuronal maturation. A morphological analysis of primary neurons confirmed that trehalose is required for neuronal arborization.
Assuntos
Astrócitos/enzimologia , Hipocampo/enzimologia , Plasticidade Neuronal/fisiologia , Neurônios/enzimologia , Trealose/biossíntese , Animais , Astrócitos/citologia , Células Cultivadas , Cerebelo/citologia , Cerebelo/enzimologia , Córtex Cerebral/citologia , Córtex Cerebral/enzimologia , Proteína Glial Fibrilar Ácida/metabolismo , Hipocampo/citologia , Masculino , Metaboloma , Metabolômica , Camundongos Endogâmicos C57BL , Microglia/citologia , Microglia/enzimologia , Proteínas Associadas aos Microtúbulos/metabolismo , Neurônios/citologia , Bulbo Olfatório/citologia , Bulbo Olfatório/enzimologia , Ratos Sprague-Dawley , Ratos WistarRESUMO
Alzheimer's disease (AD) involves the progressive degeneration of neurons critical for learning and memory. In addition, patients with AD typically exhibit impaired olfaction associated with neuronal degeneration in the olfactory bulb (OB). Because DNA base excision repair (BER) is reduced in brain cells during normal aging and AD, we determined whether inefficient BER due to reduced DNA polymerase-ß (Polß) levels renders OB neurons vulnerable to degeneration in the 3xTgAD mouse model of AD. We interrogated OB histopathology and olfactory function in wild-type and 3xTgAD mice with normal or reduced Polß levels. Compared to wild-type control mice, Polß heterozygous (Polß+/- ), and 3xTgAD mice, 3xTgAD/Polß+/- mice exhibited impaired performance in a buried food test of olfaction. Polß deficiency did not affect the proliferation of OB neural progenitor cells in the subventricular zone. However, numbers of newly generated neurons were reduced by approximately 25% in Polß+/- and 3xTgAD mice, and by over 60% in the 3xTgAD/Polß+/- mice compared to wild-type control mice. Analyses of DNA damage and apoptosis revealed significantly greater degeneration of OB neurons in 3xTgAD/Polß+/- mice compared to 3xTgAD mice. Levels of amyloid ß-peptide (Aß) accumulation in the OB were similar in 3xTgAD and 3xTgAD/Polß+/- mice, and cultured Polß-deficient neurons exhibited increased vulnerability to Aß-induced death. Olfactory deficit is an early sign in human AD, but the mechanism is not yet understood. Our findings in a new AD mouse model demonstrate that diminution of BER can endanger OB neurons, and suggest a mechanism underlying early olfactory impairment in AD.
Assuntos
Doença de Alzheimer/enzimologia , Doença de Alzheimer/patologia , DNA Polimerase beta/metabolismo , Bulbo Olfatório/enzimologia , Bulbo Olfatório/patologia , Olfato , Peptídeos beta-Amiloides/metabolismo , Animais , Apoptose , Morte Celular , Diferenciação Celular , Respiração Celular , Dano ao DNA , Modelos Animais de Doenças , Embrião de Mamíferos/metabolismo , Haploinsuficiência , Heterozigoto , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Mitocôndrias/metabolismo , Neurogênese , Neurônios/metabolismo , Neurônios/patologiaRESUMO
Herein, we assessed in a particular glial cell type, called olfactory ensheathing cells (OECs), the effect of some growth factors (GFs) on tissue transglutaminase (TG2) overexpression induced by amyloid-beta (Aß) with native full-length peptide 1-42 or by fragments, 25-35 or 35-25, as control. Previously, we demonstrated that TG2 overexpression induced by some stressors was down-regulated by GFs exposure in OECs. To monitor cell viability, an MTT test was used, while TG2 expression was examined using immunocytochemical and Western blot analysis. We also considered the involvement of the TG2-mediated apoptotic pathway. Vimentin expression was evaluated as well. Reactive oxygen species and reduced glutathione levels were utilized to test the oxidative intracellular status. Lactate dehydrogenase released into the medium, as a marker of necrotic cell death, was evaluated. We found that in OECs exposed to Aß(1-42) or Aß(25-35) for 24 h, TG2 expression increased, and we observed that the protein appeared prevalently localized in the cytosol. The pre-treatment with GFs, basic fibroblast growth factor (bFGF) or glial-derived neurotrophic factor (GDNF), down-regulated the TG2 level, which was prevalently limited to the nuclear compartment. Vimentin expression and caspase cleavage showed a significant enhancement in Aß(1-42) and Aß(25-35) exposed cells. The pre-treatment with bFGF or GDNF was able to restore the levels of the proteins to control values, and the intracellular oxidative status modified by the exposure to Aß(1-42) or Aß(25-35). Our data suggest that both bFGF or GDNF could be an innovative mechanism to contrast TG2 expression, which plays a key role in Alzheimer's disease.
Assuntos
Peptídeos beta-Amiloides/toxicidade , Proteínas de Ligação ao GTP/biossíntese , Regulação Enzimológica da Expressão Gênica , Peptídeos e Proteínas de Sinalização Intercelular/farmacologia , Bulbo Olfatório/enzimologia , Fragmentos de Peptídeos/toxicidade , Transglutaminases/biossíntese , Animais , Animais Recém-Nascidos , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/fisiologia , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Células Cultivadas , Proteínas de Ligação ao GTP/genética , Bulbo Olfatório/citologia , Bulbo Olfatório/efeitos dos fármacos , Proteína 2 Glutamina gama-Glutamiltransferase , Ratos , Ratos Wistar , Transglutaminases/genéticaRESUMO
Abnormally elevated hippocampal Caspase-6 (Casp6) activity is intimately associated with age-related cognitive impairment in humans and in mice. In humans, these high levels of Casp6 activity are initially localized in the entorhinal cortex, the area of the brain first affected by the formation of neurofibrillary tangles, according to Braak staging. The reason for the high vulnerability of entorhinal cortex neurons to neurofibrillary tangle pathology and Casp6 activity is unknown. Casp6 activity is involved in axonal degeneration, therefore, one possibility to explain increased vulnerability of the entorhinal cortex neurons would be that the afferent neurons of the olfactory bulb, some of which project their axons to the entorhinal cortex, are equally degenerating. To examine this possibility, we examined the presence of Casp6 activity, neurofibrillary tangle formation and amyloid deposition by immunohistochemistry with neoepitope antisera against the p20 subunit of active Casp6 and Tau cleaved by Casp6 (Tau∆Casp6), phosphorylated Tau paired helical filament (PHF-1) antibodies and anti-ß-amyloid antiserum, respectively, in brains from individuals with no or mild cognitive impairment and Alzheimer disease (AD) dementia. Co-localization of Casp6 activity, PHF-1 and ß-amyloid was detected mostly in the anterior olfactory nucleus (AON) of the olfactory bulb. The levels of active Casp6 in the AON, which were the highest in the AD brains, correlated with PHF-1 levels, but not with ß-amyloid levels. AON Tau∆Casp6 levels correlated with entorhinal cortex Casp6 activity and PHF-1 levels. Multiple regression analyses demonstrated that AON Casp6 activity was associated with lower global cognitive function, mini mental state exam, episodic memory and semantic memory scores. These results suggest that AON Casp6 activity could lead to Casp6-mediated degeneration in the entorhinal cortex, but cannot exclude the possibilities that entorhinal cortex degeneration signals degeneration in the AON or that the pathologies occur in both regions independently. Nevertheless, AON Casp6 activity reflects that of the entorhinal cortex.
Assuntos
Doença de Alzheimer/enzimologia , Caspase 6/metabolismo , Disfunção Cognitiva/enzimologia , Bulbo Olfatório/enzimologia , Córtex Olfatório/enzimologia , Idoso de 80 Anos ou mais , Doença de Alzheimer/patologia , Peptídeos beta-Amiloides/metabolismo , Disfunção Cognitiva/patologia , Proteínas de Ligação a DNA/metabolismo , Feminino , Hipocampo/metabolismo , Hipocampo/patologia , Humanos , Imuno-Histoquímica , Masculino , Memória/fisiologia , Entrevista Psiquiátrica Padronizada , Testes Neuropsicológicos , Bulbo Olfatório/patologia , Córtex Olfatório/patologia , Proteínas do Grupo Polycomb/metabolismo , Análise de Regressão , Índice de Gravidade de Doença , Proteínas tau/metabolismoRESUMO
Protein tyrosine phosphate δ (PTPδ), one of the receptor type IIa protein tyrosine phosphates, is known for its roles in axon guidance, synapse formation, cell adhesion, and tumor suppression. Alternative splicing of this gene generates at least four (A-D) isoforms; however, the major isoform in vivo is yet to be determined. The protein localization has neither been revealed. We have generated anti-mouse PTPδ-specific monoclonal antibody and analyzed the protein expression in wild-type and Ptpδ knockout mice. Immunoblot analysis of various organs revealed that neuronal tissues express both C-and D-isoforms of PTPδ, whereas non-neuronal tissues express only C-isoform. Immunohistochemistry of wild-type or Ptpδ heterozygous sections showed that olfactory bulb, cerebral cortex, hippocampus, cerebellum, and several nuclei in brain stem exhibit moderate to strong positive signals. These signals were absent in Ptpδ knockout specimens. Higher magnification revealed differences between expression patterns of PTPδ mRNA and its protein product. In hippocampus, weak mRNA expression in CA1 stratum pyramidale but strong immunostaining in the stratum lacunosum moleculare was observed, suggesting the axonal expression of PTPδ in the entorhinal cortical afferents. Olfactory mitral cells exhibited mRNA expression in cell bodies and protein localization in their dendritic fields, glomerular and external plexiform layers. Nissl staining showed that the external plexiform layer was reduced in Ptpδ knockout mice. Golgi-impregnation confirmed the poor dendritic growth of homozygous mitral cells. These results suggest that PTPδ may localize in axons as well as in dendrites to regulate their elaboration in the central nervous system.
Assuntos
Encéfalo/enzimologia , Neurônios/enzimologia , Proteínas Tirosina Fosfatases Classe 2 Semelhantes a Receptores/metabolismo , Animais , Anticorpos Monoclonais/administração & dosagem , Encéfalo/imunologia , Dendritos/enzimologia , Isoenzimas/metabolismo , Masculino , Camundongos , Camundongos Knockout , Bulbo Olfatório/citologia , Bulbo Olfatório/enzimologia , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Proteínas Tirosina Fosfatases Classe 2 Semelhantes a Receptores/genética , Proteínas Tirosina Fosfatases Classe 2 Semelhantes a Receptores/imunologiaRESUMO
Olfactory dysfunction is a recognized risk factor for the pathogenesis of Alzheimer's disease (AD), while the mechanisms are still not clear. Here, we applied bilateral olfactory bulbectomy (OBX), an olfactory deprivation surgery to cause permanent anosmia, in human tau-overexpressed mice (htau mice) to investigate changes of AD-like pathologies including aggregation of abnormally phosphorylated tau and cholinergic neuron loss. We found that tau phosphorylation in hippocampus was increased at Thr-205, Ser-214, Thr-231, and Ser-396 after OBX. OBX also increased the level of sarkosyl-insoluble Tau at those epitopes and accelerated accumulation of somatodendritic tau. Moreover, OBX resulted in the elevation of calpain activity accompanied by an increased expression of the cyclin-dependent kinase 5 (cdk5) neuronal activators, p35 and p25, in hippocampus. Furthermore, OBX induces the loss of the cholinergic neurons in medial septal. Administration of cdk5 pharmacological inhibitor roscovitine into lateral ventricles suppressed tau hyperphosphorylation and mislocalization and restored the cholinergic neuron loss. These findings suggest that olfactory deprivation by OBX hastens tau pathology and cholinergic system impairment in htau mice possibly via activation of cdk5.
Assuntos
Doença de Alzheimer/enzimologia , Doença de Alzheimer/patologia , Quinase 5 Dependente de Ciclina/metabolismo , Bulbo Olfatório/enzimologia , Bulbo Olfatório/patologia , Proteínas tau/metabolismo , Animais , Calpaína/metabolismo , Neurônios Colinérgicos/metabolismo , Dendritos/efeitos dos fármacos , Dendritos/metabolismo , Modelos Animais de Doenças , Ativação Enzimática/efeitos dos fármacos , Humanos , Camundongos , Bulbo Olfatório/efeitos dos fármacos , Bulbo Olfatório/cirurgia , Fosforilação/efeitos dos fármacos , Purinas/farmacologia , Roscovitina , Núcleos Septais/efeitos dos fármacos , Núcleos Septais/metabolismo , Solubilidade , Tauopatias/metabolismo , Tauopatias/patologiaRESUMO
INTRODUCTION: Dopamine is one of the major brain neurotransmitters, and the loss of dopaminergic neurons in basal ganglia cause motor deficits in Parkinson's disease. We proposed that the difficulty in olfaction observed in the elderly may be due to an alteration in the number of dopaminergic neurons. MATERIALS AND METHODS: Sections were taken from olfactory bulbs of post-mortem tissue specimens of 13 humans, males and females, aged from 19 to 63 years (≤35 and ≥50 years), with no history of neurological disorders. The tissues were fixed, embedded, cut on a freezing microtome, and prepared for immunohistochemical analysis using tyrosine hydroxylase (TH) and aromatic l-amino acid decarboxylase (AADC) antibodies. The number of positive neurons was counted. RESULTS: TH- and AADC-positive cells were present in the glomerular layer. There was no significant difference between the numbers of TH- and AADC-positive cells, in males and females, and in young and elderly individuals. The quantitative analysis revealed that the number of TH- and AADC-positive neurons were significantly higher in males than in females (P<0.05). Moreover, there was a significant increase in the number of TH- and AADC-positive neurons in the olfactory bulbs of the elderly compared with young individuals (P<0.05). CONCLUSION: Factors such as gender and age may affect the number of dopaminergic neurons, and there is a correlation between increased dopaminergic neurons and olfactory performance. Moreover, the increase in dopaminergic cells in the olfactory bulb of the elderly may indicate the existence of rostral migratory stream in adult humans.
Assuntos
Dopamina/fisiologia , Neurônios Dopaminérgicos/citologia , Bulbo Olfatório/citologia , Olfato , Adulto , Fatores Etários , Descarboxilases de Aminoácido-L-Aromático/análise , Descarboxilases de Aminoácido-L-Aromático/metabolismo , Contagem de Células , Dopamina/metabolismo , Neurônios Dopaminérgicos/química , Neurônios Dopaminérgicos/enzimologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Bulbo Olfatório/química , Bulbo Olfatório/enzimologia , Fatores Sexuais , Tirosina 3-Mono-Oxigenase/análise , Tirosina 3-Mono-Oxigenase/metabolismo , Adulto JovemRESUMO
Dopamine (DA) inhibits, whereas gonadotrophin-releasing hormone (GnRH) stimulates, luteinisiing (LH) cells in the pituitary of some but not all teleosts. A reduction in the hypophysiotropic dopaminergic tone is necessary for the stimulatory effect of GnRH on LH cells. Neuropeptide Y (NPY) has emerged as one of the potent, endogenous agent that modulates LH secretion directly or indirectly via GnRH. Involvement of NPY in the regulation of hypophysiotropic DA neurones, however, is not known, but there is good evidence suggesting an interaction in the mammalian hypothalamus. DA neurones, identified by tyrosine hydroxylase (TH)-immunoreactivity, were observed widely throughout the brain of the Indian major carp, Cirrhinus cirrhosus. The granule cells and ganglion cells of terminal nerve in the olfactory bulb, and cells in ventral telencephalon and preoptic area (POA) showed conspicuous TH immunoreactivity. In the POA, the nucleus preopticus periventricularis (NPP), divisible into anterior (NPPa) and posterior (NPPp) components, showed prominent TH-immunoreactivity. The majority of TH neurones in NPPa showed axonal extensions to the pituitary and were closely associated with LH cells. The NPPa also appeared to be the site for intense interaction between NPY and DA because it contains a rich network of NPY fibres and few immunoreactive cells. Approximately 89.7 ± 1.5% TH neurones in NPPa were contacted by NPY fibres. Superfused POA slices treated with a NPY Y2 -receptor agonist, NPY 13-36 resulted in a significant (P < 0.001) reduction in TH-immunoreactivity in NPPa. TH neurones in NPPa did not respond to NPY Y1 -receptor agonist, [Leu(31) , Pro(34) ] Neuropeptide Y treatment. We suggest that, by inhibiting DAergic neurones in NPPa via Y2 -receptors, NPY may contribute to the up-regulation of the GnRH-LH cells axis. The microcircuitry of DA and NPY and their interaction in NPPa might be a crucial component in the central regulation of LH secretion in the teleosts.
Assuntos
Carpas/fisiologia , Neuropeptídeo Y/fisiologia , Bulbo Olfatório/enzimologia , Hipófise/enzimologia , Área Pré-Óptica/fisiologia , Prosencéfalo/enzimologia , Tirosina 3-Mono-Oxigenase/fisiologia , Animais , Feminino , Telencéfalo/enzimologiaRESUMO
Akt, also known as protein kinase B, plays key roles in cell proliferation, survival and metabolism. Akt hyperactivation contributes to many pathophysiological conditions, including human cancers, and is closely associated with poor prognosis and chemo- or radiotherapeutic resistance. Phosphorylation of Akt at S473 (ref. 5) and T308 (ref. 6) activates Akt. However, it remains unclear whether further mechanisms account for full Akt activation, and whether Akt hyperactivation is linked to misregulated cell cycle progression, another cancer hallmark. Here we report that Akt activity fluctuates across the cell cycle, mirroring cyclin A expression. Mechanistically, phosphorylation of S477 and T479 at the Akt extreme carboxy terminus by cyclin-dependent kinase 2 (Cdk2)/cyclin A or mTORC2, under distinct physiological conditions, promotes Akt activation through facilitating, or functionally compensating for, S473 phosphorylation. Furthermore, deletion of the cyclin A2 allele in the mouse olfactory bulb leads to reduced S477/T479 phosphorylation and elevated cellular apoptosis. Notably, cyclin A2-deletion-induced cellular apoptosis in mouse embryonic stem cells is partly rescued by S477D/T479E-Akt1, supporting a physiological role for cyclin A2 in governing Akt activation. Together, the results of our study show Akt S477/T479 phosphorylation to be an essential layer of the Akt activation mechanism to regulate its physiological functions, thereby providing a new mechanistic link between aberrant cell cycle progression and Akt hyperactivation in cancer.
Assuntos
Ciclo Celular/fisiologia , Proteínas Proto-Oncogênicas c-akt/química , Proteínas Proto-Oncogênicas c-akt/metabolismo , Animais , Apoptose/genética , Proliferação de Células , Ciclina A2/metabolismo , Quinase 2 Dependente de Ciclina/metabolismo , Células-Tronco Embrionárias/citologia , Células-Tronco Embrionárias/metabolismo , Ativação Enzimática , Masculino , Alvo Mecanístico do Complexo 2 de Rapamicina , Camundongos , Complexos Multiproteicos/metabolismo , Neoplasias/enzimologia , Neoplasias/patologia , Bulbo Olfatório/citologia , Bulbo Olfatório/enzimologia , Bulbo Olfatório/metabolismo , Proteína Oncogênica v-akt/química , Proteína Oncogênica v-akt/metabolismo , Fosforilação , Fosfosserina/metabolismo , Fosfotreonina/metabolismo , Serina-Treonina Quinases TOR/metabolismoRESUMO
Melatonin is a neurohormone associated with circadian rhythms. A diurnal rhythm in olfactory sensitivity has been previously reported and melatonin receptor mRNAs have been observed in the olfactory bulb, but the effects of melatonin in the olfactory bulb have not been explored. First, we corroborated data from a previous study that identified melatonin receptor messenger RNAs in the olfactory bulb. We then investigated whether melatonin treatment would affect cells in the olfactory bulbs of rats. Using a combination of polymerase chain reaction (PCR), quantitative PCR (qPCR), cell culture, and electrophysiology, we discovered that melatonin receptors and melatonin synthesis enzymes were present in the olfactory bulb and we observed changes in connexin43 protein, GluR1 mRNA, GluR2 mRNA, Per1 mRNA, Cry2 mRNA, and K(+) currents in response to 2-iodomelatonin. Via qPCR, we observed that messenger RNAs encoding melatonin receptors and melatonin biosynthesis enzymes fluctuated in the olfactory bulb across 24h. Together, these data show that melatonin receptors are present in the olfactory bulb and likely affect olfactory function. Additionally, these data suggest that melatonin may be locally synthesized in the olfactory bulb.
Assuntos
Melatonina/metabolismo , Bulbo Olfatório/metabolismo , Receptor MT1 de Melatonina/metabolismo , Receptor MT2 de Melatonina/metabolismo , Animais , Astrócitos/metabolismo , Células Cultivadas , Ritmo Circadiano , Feminino , Técnicas In Vitro , Masculino , Camundongos , Camundongos Knockout , Neurônios/metabolismo , Bulbo Olfatório/enzimologia , Potássio/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Receptor MT1 de Melatonina/genética , Receptor MT2 de Melatonina/genéticaRESUMO
Compelling evidence from animal models and clinical studies suggest that transplantation of olfactory ensheathing cells (OECs), specialized glia in the olfactory system, combined with specific training may be therapeutically useful in the central nervous system (CNS) injuries and neurodegenerative diseases. The unique function of OECs could mainly attribute to both production of cell adhesion molecules and secretion of growth factors in OECs, which support neuron survival and neurite outgrowth. However, little is known about whether engulfment of neuronal degenerative debris by OECs also equally contributes to neuronal survival and neurite outgrowth. Furthermore, the molecular mechanisms responsible for neuronal degenerative corpses' removal remain elusive. Here, we used an in vitro model of primary culture of spinal cord neurons to investigate the effect of engulfment of degenerative neuron debris by OECs on neuronal survival and neurite outgrowth and the possible molecular mechanisms. Our results showed that OECs can engulf an amount of degenerated neuron debris, and this phagocytosis can make a substantial contribution to neuron growth, as demonstrated by increased number of neurons with longer neurite length and richer neurite branches when compared with the combination of neuron debris and OEC conditioned medium (OECCM). Moreover, p38 mitogen-activated protein kinase (p38MAPK) signaling pathway may mediate the OEC engulfment of debris because the p38MAPK-specific inhibitor, SB203580, can abrogate all the positive effects of OECs, including clearance of degenerated neuron debris and generation of bioactive molecules, indicating that p38MAPK is required for the process of phagocytosis of the neuron debris. In addition, the OEC phagocytic activity had no influence on its generation of bioactive molecules. Therefore, these findings provide new insight into further investigations on the OEC role in the repair of traumatic CNS injury and neurodegenerative diseases.
Assuntos
Citofagocitose/fisiologia , Neuritos/enzimologia , Bulbo Olfatório/citologia , Bulbo Olfatório/enzimologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Sobrevivência Celular/fisiologia , Ativação Enzimática/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Neurônios/enzimologiaRESUMO
The microscopic organization of the piriform cortex (PC) was studied in normal and experimental material from adult albino rats. In rapid-Golgi specimens a set of collaterals from the lateral olfactory tract (i.e., sublayer Ia) to the neuropil of the Layer II (LII) was identified. Specimens from experimental animals that received electrolytic lesion of the main olfactory bulb three days before sacrificing, were further processed for pre-embedding immunocytochemistry to the enzyme glutamic acid decarboxylase 67 (GAD 67). This novel approach permitted a simultaneous visualization at electron microscopy of both synaptic degeneration and GAD67-immunoreactive (GAD-I) sites. Degenerating and GAD-I synapses were separately found in the neuropil of Layers I and II of the PC. Previously overlooked patches of neuropil were featured in sublayer Ia. These areas consisted of dendritic and axonal processes including four synaptic types. Tridimensional reconstructions from serial thin sections from LI revealed the external appearance of the varicose and tubular dendrites as well as the synaptic terminals therein. The putative source(s) of processes to the neuropil of sublayer Ia is discussed in the context of the internal circuitry of the PC and an alternative model is introduced.
Assuntos
Neurópilo/ultraestrutura , Condutos Olfatórios/ultraestrutura , Animais , Biomarcadores/análise , Eletrólise , Feminino , Glutamato Descarboxilase/análise , Imuno-Histoquímica , Masculino , Microscopia Eletrônica , Rede Nervosa/enzimologia , Rede Nervosa/ultraestrutura , Técnicas de Rastreamento Neuroanatômico , Neurópilo/enzimologia , Bulbo Olfatório/enzimologia , Bulbo Olfatório/lesões , Bulbo Olfatório/ultraestrutura , Condutos Olfatórios/enzimologia , Condutos Olfatórios/lesões , Ratos , Ratos Sprague-Dawley , Ratos WistarRESUMO
Nonmuscle myosin II (NM II) is the name given to the multi-subunit protein product of three genes (myh9, myh10, and myh14) encoding different nonmuscle myosin heavy chains. The three NM II isoforms share a very similar molecular structure and play important roles in a variety of fundamental biological processes. NM II-B (myh10) has been shown to be essential for the formation of mouse neural system and heart. But so far the complete knowledge for its expression in developing zebrafish embryos is lacking. In current study, we proved the conservation of zebrafish NM II-B in vertebrate evolution by in silicon analysis. Afterwards the NM II-B (myh10) expression was demonstrated to initiate after gastrulation stage. At 20 hpf, the expression is mainly restricted in central nervous system (CNS). It was maintained and expanded to sensor organ including eye, otic vesicle, and olfactory bulb at 36 hpf and later. We also detected myh10 mRNA hybridization signal in 48 hpf zebrafish heart. In addition, we investigated myh9a and myh9b mRNA distribution in zebrafish developing embryos. It was shown that myh10 and myh9 have distinct expression pattern, with myh9s not in neural system but in epidermis, enveloping layer (EVL). Our study provides new insight into the NM II expression and the use of this model organism to tackle future studies on the role of NM II in embryo development.
Assuntos
Sistema Nervoso Central/metabolismo , Epiderme/metabolismo , Cadeias Pesadas de Miosina/genética , Miosina não Muscular Tipo IIB/genética , Peixe-Zebra/embriologia , Sequência de Aminoácidos , Animais , Sistema Nervoso Central/citologia , Sistema Nervoso Central/embriologia , Embrião não Mamífero/metabolismo , Desenvolvimento Embrionário/genética , Células Epidérmicas , Epiderme/embriologia , Regulação da Expressão Gênica no Desenvolvimento , Coração/embriologia , Coração/fisiologia , Humanos , Hibridização in Situ Fluorescente , Camundongos , Dados de Sequência Molecular , Cadeias Pesadas de Miosina/química , Cadeias Pesadas de Miosina/metabolismo , Miosina não Muscular Tipo IIB/química , Miosina não Muscular Tipo IIB/metabolismo , Bulbo Olfatório/citologia , Bulbo Olfatório/enzimologia , Bulbo Olfatório/metabolismo , Filogenia , Ratos , Alinhamento de Sequência , Peixe-Zebra/genética , Peixe-Zebra/metabolismoRESUMO
Endothelins (ETs) are widely expressed in the olfactory bulb (OB) and other brain areas where they function as neuropeptides. In a previous study we reported that in the OB ET-1 and ET-3 participate in the long-term regulation of tyrosine hydroxylase (TH), the key enzyme in catecholamine biosynthesis. ETs stimulate TH activity by increasing total and phosphorylated enzyme levels as well as its mRNA. ET-1 response is mediated by a super high affinity ETA receptor coupled to adenylyl cyclase/protein kinase A and Ca(2+)/calmodulin-dependent protein kinase II (CaMK-II) activation whereas that of ET-3 through an atypical receptor coupled not only to these signaling pathways but also to phospholipase C (PLC)/protein kinase C pathway. Given the participation of PLC and CaMKII in the regulation of TH by ETs in the OB we sought to establish the contribution of calcium to ETs response. Present findings show that calcium released from ryanodine-sensitive channels and extracellular calcium were necessary to stimulate TH by ETs through CaMK-II. On the other hand, intracellular calcium released by the endoplasmic reticulum partially mediated ETs-evoked increase in TH mRNA but calcium influx and CaMK-II inhibition abolished the response. However calcium mechanisms were not involved in ETs-evoked increase in TH protein content. Present findings support that different sources of calcium contribute to the long-term modulation of TH activity and expression mediated by ETs in the rat OB.
Assuntos
Cálcio/metabolismo , Endotelinas/fisiologia , Bulbo Olfatório/enzimologia , Tirosina 3-Mono-Oxigenase/metabolismo , Animais , Sequência de Bases , Primers do DNA , Masculino , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Tirosina 3-Mono-Oxigenase/genéticaRESUMO
Thioether-containing pesticides are more toxic in certain anadromous and catadromous fish species that have undergone acclimation to hypersaline environments. Enhanced toxicity has been shown to be mediated through the bioactivation of these xenobiotics by one or more flavin-containing monooxygenases (FMOs), which are induced by hyperosmotic conditions. To better understand the number of FMO genes that may be regulated by hyperosmotic conditions, rainbow trout (Oncorhynchus mykiss) were maintained and acclimated to freshwater (<0.5 g/L salinity) and to 18 g/L salinity. The expression of 3 different FMO transcripts (A, B and C) and associated enzymatic activities methyl p-tolyl sulfoxidation (MTSO) and benzydamine N-oxigenation (BZNO) were measured in four tissues. In freshwater-acclimated organisms FMO catalytic activities were as follows: liver>kidney>gills=olfactory tissues; in hypersaline-acclimated animals activities were higher in liver>gills>olfactory tissues>kidney. Acclimation to 18 g/L caused a significant induction in the stereoselective formation of R-MTSO in gill. In olfactory tissues, stereoselective (100%) formation of S-MTSO was observed and was unaltered by acclimation to hypersaline water. When specific transcripts were evaluated, salinity-acclimation increased FMO A in liver (up to 2-fold) and kidney (up to 3-fold) but not in olfactory tissues and gills. FMO B mRNA was significantly down-regulated in all tissues, and FMO C was unchanged by hypersaline acclimation. FMO B and C failed to correlate with any FMO catalytic activity, but FMO A mRNA expression linearly correlated to both FMO catalytic activities (MTSO and BZNO) in liver (r(2)=0.92 and r(2)=0.88) and kidney microsomes (r(2)=0.93 and r(2)=90). FMO A only correlated with MTSO activity in gills (r(2)=0.93). These results indicate unique tissue specific expression of FMO genes in salmonids and are consistent with salinity-mediated enhancement of thioether-containing pesticide bioactivation by FMO which may occur in liver or kidney after salinity acclimation.