RESUMO
In Parkinson's disease (PD), progressive degeneration of nigrostriatal innervation leads to atrophy and loss of dendritic spines of striatal medium spiny neurons (MSNs). The loss disrupts corticostriatal transmission, impairs motor behavior, and produces nonmotor symptoms. Nigral neurons express brain-derived neurotropic factor (BDNF) and dopamine D3 receptors, both protecting the dopamine neurons and the spines of MSNs. To restore motor and nonmotor symptoms to normality, we assessed a combined therapy in a bilateral rat Parkinson's model, with only 30% of surviving neurons. The preferential D3 agonist pramipexole (PPX) was infused for four ½ months via mini-osmotic pumps and one month after PPX initiation; the BDNF-gene was transfected into the surviving nigral cells using the nonviral transfection NTS-polyplex vector. Overexpression of the BDNF-gene associated with continuous PPX infusion restored motor coordination, balance, normal gait, and working memory. Recovery was also related to the restoration of the average number of dendritic spines of the striatal projection neurons and the number of TH-positive neurons of the substantia nigra and ventral tegmental area. These positive results could pave the way for further clinical research into this promising therapy.
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In Alzheimer's disease (AD), two mutually exclusive amino-terminal-dependent conformations have been reported to occur during the aggregation of Tau protein into neurofibrillary tangles (NFTs). An early conformation of full-length Tau, involving the bending of the amino terminus over the third repeated domain, is recognized by the Alz-50 antibody, followed by a second conformation recognized by Tau-66 antibody that depends on the folding of the proline-rich region over the third repeated domain in a molecule partially truncated at the amino- and carboxyl-termini. α-1-antichymotrypsin (ACT) is an acute phase serum glycoprotein that accumulates abnormally in the brain of AD patients, and since it is considered to promote the in vitro and in vivo aggregation of amyloid-ß, we here seek further evidence that ACT may also contribute to the abnormal aggregation of Tau in AD. By analyzing brain samples from a population of AD cases under immunofluorescence and high-resolution confocal microscopy, we demonstrate here the abundant expression of ACT in hippocampal neurons, visualized as a granular diffuse accumulation, frequently reaching the nuclear compartment. In a significant number of these neurons, intracellular NFTs composed of abnormally phosphorylated and truncated Tau at Asp421 were also observed to coexist in separated regions of the cytoplasm. However, we found strong colocalization between ACT and diffuse aggregates of Tau-66-positive granules, which was not observed with Alz-50 antibody. These results suggest that ACT may play a role during the development of Tau conformational changes facilitating its aggregation during the formation of the neurofibrillary pathology in AD.
Assuntos
Doença de Alzheimer , Humanos , Doença de Alzheimer/metabolismo , Proteínas tau/metabolismo , Emaranhados Neurofibrilares/metabolismo , Emaranhados Neurofibrilares/patologia , Neurônios/metabolismo , Encéfalo/metabolismo , AnticorposRESUMO
Several types of sensory perception have circadian rhythms. The spinal cord can be considered a center for controlling circadian rhythms by changing clock gene expression. However, to date, it is not known if mechanonociception itself has a circadian rhythm. The hypothalamic A11 area represents the primary source of dopamine (DA) in the spinal cord and has been found to be involved in clock gene expression and circadian rhythmicity. Here, we investigate if the paw withdrawal threshold (PWT) has a circadian rhythm, as well as the role of the dopaminergic A11 nucleus, DA, and DA receptors (DR) in the PWT circadian rhythm and if they modify clock gene expression in the lumbar spinal cord. Naïve rats showed a circadian rhythm of the PWT of almost 24 h, beginning during the night-day interphase and peaking at 14.63 h. Similarly, DA and DOPAC's spinal contents increased at dusk and reached their maximum contents at noon. The injection of 6-hydroxydopamine (6-OHDA) into the A11 nucleus completely abolished the circadian rhythm of the PWT, reduced DA tissue content in the lumbar spinal cord, and induced tactile allodynia. Likewise, the repeated intrathecal administration of D1-like and D2-like DA receptor antagonists blunted the circadian rhythm of PWT. 6-OHDA reduced the expression of Clock and Per1 and increased Per2 gene expression during the day. In contrast, 6-OHDA diminished Clock, Bmal, Per1, Per2, Per3, Cry1, and Cry2 at night. The repeated intrathecal administration of the D1-like antagonist (SCH-23390) reduced clock genes throughout the day (Clock and Per2) and throughout the night (Clock, Per2 and Cry1), whereas it increased Bmal and Per1 throughout the day. In contrast, the intrathecal injection of the D2 receptor antagonists (L-741,626) increased the clock genes Bmal, Per2, and Per3 and decreased Per1 throughout the day. This study provides evidence that the circadian rhythm of the PWT results from the descending dopaminergic modulation of spinal clock genes induced by the differential activation of spinal DR.
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Amyloid-beta oligomers (AßO) have been proposed as the most potent neurotoxic and inflammation inducers in Alzheimer's disease (AD). AßO contribute to AD pathogenesis by impairing the production of several cytokines and inflammation-related signaling pathways, such as the Janus kinases/signal transducer of transcription factor-3 (JAK/STAT3) pathway. STAT3 modulates glial activation, indirectly regulates Aß deposition, and induces cognitive decline in AD transgenic models. However, in vivo studies using an AßO microinjection rat model have not yet explored STAT3 role. The main purpose of this study was to elucidate if a single microinjection of AßO could promote an increased expression of STAT3 in glial cells favoring neuroinflammation and neurodegeneration. We designed a model of intrahippocampal microinjection and assessed glial activation, cytokines production, STAT3 expression, and neurodegeneration in time. Our results showed robust expression of STAT3 in glial cells (mainly in astrocytes) and neurons, correlating with neuronal death in response to AßO administration. A STAT3 inhibition assay conducted in rat primary hippocampal cultures, suggested that the induction of the transcription factor by AßO in astrocytes leads them to an activation state that may favor neuronal death. Notwithstanding, pharmacological inhibition of the JAK2/STAT3 pathway should be focused on astrocytes because it is also essential in neurons survival. Overall, these findings strongly suggest the participation of STAT3 in the development of neurodegeneration.
Assuntos
Peptídeos beta-Amiloides/metabolismo , Astrócitos/metabolismo , Gliose/etiologia , Gliose/metabolismo , Neurônios/metabolismo , Fator de Transcrição STAT3/metabolismo , Doença de Alzheimer/etiologia , Doença de Alzheimer/metabolismo , Doença de Alzheimer/patologia , Animais , Astrócitos/patologia , Biomarcadores , Morte Celular , Modelos Animais de Doenças , Suscetibilidade a Doenças , Imunofluorescência , Gliose/patologia , Hipocampo/metabolismo , Hipocampo/patologia , Imuno-Histoquímica , Agregados Proteicos , Agregação Patológica de Proteínas/genética , Agregação Patológica de Proteínas/metabolismo , Multimerização Proteica , Ratos , Fator de Transcrição STAT3/genéticaRESUMO
In Alzheimer's disease, tau protein undergoes post-translational modifications including hyperphosphorylation and truncation, which promotes two major conformational changes associated with progressive N-terminal folding. Along with the development of the disease, tau ubiquitination was previously shown to emerge in the early and intermediate stages of the disease, which is closely associated with early tau truncation at aspartic acid 421, but not with a subsequently truncated tau molecule at glutamic acid 391. In the same group of cases, using multiple immunolabeling and confocal microscopy, a possible relationship between the ubiquitin-targeting of tau and the progression of conformational changes adopted by the N-terminus of this molecule was further studied. A comparable number of neurofibrillary tangles was found displaying ubiquitin, an early conformation recognized by the Alz-50 antibody, and a phosphorylation. However, a more reduced number of neurofibrillary tangles were immunoreactive to Tau-66 antibody, a late tau conformational change marker. When double-labeling profiles of neurofibrillary tangles were assessed, ubiquitination was clearly demonstrated in tau molecules undergoing early N-terminal folding, but was barely observed in late conformational changes of the N-terminus adopted by tau. The same pattern of colocalization was visualized in neuritic pathology. Overall, these results indicate that a more intact conformation of the N-terminus of tau may facilitate tau ubiquitination, but this modification may not occur in a late truncated and more compressed folding of the N-terminus of the tau molecule.
Assuntos
Doença de Alzheimer/patologia , Encéfalo/patologia , Emaranhados Neurofibrilares/química , Ubiquitinação/fisiologia , Proteínas tau/química , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/metabolismo , Encéfalo/metabolismo , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Emaranhados Neurofibrilares/patologia , Conformação Proteica , Proteínas tau/metabolismoRESUMO
We previously demonstrated that, in the early stages of tau processing in Alzheimer's disease, the N-terminal part of the molecule undergoes a characteristic cascade of phosphorylation and progressive misfolding of the proteins resulting in a structural conformation detected by Alz-50. In this immunohistochemical study of AD brain tissue, we have found that C-terminal truncation of tau at Asp-421 was an early event in tau aggregation and analyzed the relationship between phospho-dependent tau epitopes located at the C-terminus with truncation at Glu-391. The aim of this study was to determine whether C-terminal truncation may trigger events leading to the assembly of insoluble PHFs from soluble tau aggregates present in pre-tangle cells. Our findings suggest that there is a complex interaction between phosphorylated and truncated tau species. A model is presented here in which truncated tau protein represents an early neurotoxic species while phosphorylated tau species may provide a neuroprotective role in Alzheimer's disease.
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Obesity is a serious worldwide health problem, affecting 20-40% of the population in several countries. According to animal models, obesity is related to changes in the expression of proteins that control energy homeostasis and in neurotransmission associated to regulation of food intake. For example, it has been reported that diet-induced obesity produces overexpression of dopamine D4 receptor (D4R) mRNA in the ventromedial hypothalamic nucleus (VMH) of mice. Neurons in the VMH send dense glutamatergic projections to other hypothalamic regions as the paraventricular nucleus (PVN), where multiple signals are integrated to finely regulate energy homeostasis and food intake. Although it is well established that dopaminergic transmission in the hypothalamus plays a key role in modulating feeding, the specific mechanisms involved in the activation of D4R in the PVN and its modulatory action on glutamate release and feeding behavior have remained unexplored. To fill this gap, we characterize the behavioral and neurochemical role of D4R in the PVN. In behavioral experiments, we examined the effects of activation of dopamine D4 receptors in the PVN on food intake and on the behavioral satiety sequence in rats exposed to a food-restricted feeding program. In vitro experiments were conducted to study the effects of activation of dopamine D4 receptors on [(3)H]glutamate release and on plasma corticosterone in explants of the PVN. We found that activation of D4R in the PVN induced inhibition of glutamate release and stimulated food intake by inhibiting satiety. Furthermore, activation of D4R in the PVN decreased plasma levels of corticosterone, and this effect was reverted by NMDA. According to our findings, D4R in the PVN may be a target for the pharmacotherapy for obesity as well as eating disorder patients who show restrictive patterns and overweight.
Assuntos
Ácido Glutâmico/metabolismo , Hiperfagia/fisiopatologia , Núcleo Hipotalâmico Paraventricular/metabolismo , Receptores de Dopamina D4/metabolismo , Animais , Benzamidas/farmacologia , Corticosterona/sangue , Agonistas de Dopamina/farmacologia , Antagonistas de Dopamina/farmacologia , Interações Medicamentosas , Ingestão de Alimentos/efeitos dos fármacos , Jejum , Agonistas de Receptores de GABA-A/farmacologia , Ácido Glutâmico/farmacologia , Hiperfagia/induzido quimicamente , Masculino , Muscimol/farmacologia , Núcleo Hipotalâmico Paraventricular/efeitos dos fármacos , Piperazinas/farmacologia , Piridinas/farmacologia , Pirimidinas/farmacologia , Pirróis/farmacologia , Ratos , Ratos Wistar , Saciação/efeitos dos fármacos , Trítio/farmacocinéticaRESUMO
The aim of this study was to determine the effect of chronic undernutrition on the content and release of γ-amino butyric acid (GABA) and glutamate (GLU) transmitters in the rat spinal cord. The release of [(3)H]-GABA and [(3)H]-GLU was determined by radioactive liquid scintillation techniques, and the concentrations of GABA and GLU in spinal cord preparations from control and undernourished young rats (50-60 days old) were measured by reverse-phase HPLC. The GABA and GLU contents in the lumbar spinal dorsal horn (L6 segment) were significantly lower in undernourished rats relative to control rats (22.2 ± 3.7 and 10.7 ± 1.9 %, respectively; P < 0.05). Spinal cord blocks from undernourished animals also showed lower rates of [(3)H]-GABA and [(3)H]-GLU release than controls (27.6 ± 3.5 and 12.8 ± 2.5 %, respectively; P < 0.01). We propose that the decreases in GLU content and release are consistent with a reduced activation of either afferent fibers, spinal glutaminergic neurons, or both. Furthermore, we propose that the decreased content and release of GABA in undernourished animals are related to a depression in pre- and post-synaptic inhibition. In addition, we hypothesize that the reductions in GABA content and release serve as compensatory mechanisms to counterbalance decreases in sensory transmission and GLU content in the spinal cord of the chronically undernourished rat.
Assuntos
Ácido Glutâmico/metabolismo , Desnutrição/metabolismo , Medula Espinal/metabolismo , Ácido gama-Aminobutírico/metabolismo , Animais , Índice de Massa Corporal , Peso Corporal/fisiologia , Cálcio/fisiologia , Interpretação Estatística de Dados , Feminino , Neurotransmissores/metabolismo , Potássio/farmacologia , Ratos , Ratos Wistar , Medula Espinal/efeitos dos fármacos , Estimulação QuímicaRESUMO
BACKGROUND: Feeding behavior is deeply affected by serotonergic neurotransmission. This regulatory activity is mediated mainly by specific 5-HT1/2 receptors, and the paraventricular nucleus (PVN) plays a key role in this phenomena. In order to reveal the involvement of 5-HT1A and 5-HT1B receptors in the paraventricular nucleus of the hypothalamus (PVN) on serotonin-induced hypophagia, we examined the effects of intra-PVN injections of serotonin in WAY 100635 or SB 216641-pretreated rats on the structure of feeding behavior. MATERIAL/METHODS: Male Wistar rats were kept at 21+/-1 degrees C with a 12 h light /dark cycle on a self-selection feeding paradigm, provided with freely available and separate sources of protein, carbohydrate, fat and water. Blockade of 5-HT1A or 5-HT1B receptors in the paraventricular nucleus was effected by WAY 100635 (2 microg) or SB-216641 (2 microg) pretreatment; ten minutes later, 5-HT (2 microg) was applied into the same nucleus, then food intake and meal patterns were measured in a 30-minute period. The behavioral test was conducted at the beginning of the dark phase. RESULTS: The suppressive effect of 5-HT on carbohydrate intake was blocked by both WAY 100635 and SB 216641 at the beginning of the active (dark) feeding period. CONCLUSIONS: The hypophagic effect induced by 5-HT requires activation of 5-HT1A and 5-HT1B receptors, and the specific contribution of these subtype receptors is different, since the 5-HT1A subtype showed higher behavioral selectivity.