61,713 research outputs found
Stimulation of microglial metabotropic glutamate receptor mGlu2 triggers tumor necrosis factor alpha-induced neurotoxicity in concert with microglial-derived fas ligand
Activated microglia may be detrimental to neuronal survival in a number of neurodegenerative diseases. Thus, strategies that reduce microglial neurotoxicity may have therapeutic benefit. Stimulation of group II metabotropic glutamate (mGlu) receptors on rat primary microglia with the specific group II agonist 2S, 2 ' R, 3 ' R- 2-(2 ', 3 '-dicarboxy-cyclopropyl) glycine for 24 h induced microglial activation and resulted in a neurotoxic microglial phenotype. These effects were attributable to preferential mGlu2 stimulation, because N-acetyl-L-aspartyl-L-glutamate, a specific mGlu3 agonist, did not induce microglial activation or neurotoxicity. Stimulation of microglial mGlu2 but not mGlu3 induced caspase-3 activation in cerebellar granule neurons in culture, using microglial-conditioned media as well as cocultures. Stimulation of microglial mGlu2 induced tumor necrosis factor-alpha(TNF alpha) release, which contributed to microglial neurotoxicity mediated via neuronal TNF receptor 1 and caspase-3 activation. Stimulation of microglial group I or III mGlu receptors did not induce TNF alpha release. TNF alpha was only neurotoxic in the presence of microglia or microglial-conditioned medium. The toxicity of TNF alpha could be prevented by coexposure of neurons to conditioned medium from microglia stimulated by the specific group III agonist L-2-amino-4-phosphono-butyric acid. The neurotoxicity of TNF alpha derived from mGlu2-stimulated microglia was potentiated by microglial-derived Fas ligand (FasL), the death receptor ligand. FasL was constitutively expressed in microglia and shed after mGlu2 stimulation. Our data suggest that selective and inverse modulation of microglial mGlu2 and mGlu3 may prove a therapeutic target in neuroinflammatory diseases such as Alzheimer's disease and multiple sclerosis
Microglial K(+) channel expression in young adult and aged mice.
The K(+) channel expression pattern of microglia strongly depends on the cells' microenvironment and has been recognized as a sensitive marker of the cells' functional state. While numerous studies have been performed on microglia in vitro, our knowledge about microglial K(+) channels and their regulation in vivo is limited. Here, we have investigated K(+) currents of microglia in striatum, neocortex and entorhinal cortex of young adult and aged mice. Although almost all microglial cells exhibited inward rectifier K(+) currents upon membrane hyperpolarization, their mean current density was significantly enhanced in aged mice compared with that determined in young adult mice. Some microglial cells additionally exhibited outward rectifier K(+) currents in response to depolarizing voltage pulses. In aged mice, microglial outward rectifier K(+) current density was significantly larger than in young adult mice due to the increased number of aged microglial cells expressing these channels. Aged dystrophic microglia exhibited outward rectifier K(+) currents more frequently than aged ramified microglia. The majority of microglial cells expressed functional BK-type, but not IK- or SK-type, Ca(2+) -activated K(+) channels, while no differences were found in their expression levels between microglia of young adult and aged mice. Neither microglial K(+) channel pattern nor K(+) channel expression levels differed markedly between the three brain regions investigated. It is concluded that age-related changes in microglial phenotype are accompanied by changes in the expression of microglial voltage-activated, but not Ca(2+) -activated, K(+) channels
Distinct Microglial Responses in Two Transgenic Murine Models of TAU Pathology
Microglial cells are crucial players in the pathological process of neurodegenerative
diseases, such as Alzheimer’s disease (AD). Microglial response in AD has been
principally studied in relation to amyloid-beta pathology but, comparatively, little is known
about inflammatory processes associated to tau pathology. In the hippocampus of
AD patients, where tau pathology is more prominent than amyloid-beta pathology,
a microglial degenerative process has been reported. In this work, we have directly
compared the microglial response in two different transgenic tau mouse models:
ThyTau22 and P301S. Surprisingly, these two models showed important differences
in the microglial profile and tau pathology. Where ThyTau22 hippocampus manifested
mild microglial activation, P301S mice exhibited a strong microglial response in parallel
with high phospho-tau accumulation. This differential phospho-tau expression could
account for the different microglial response in these two tau strains. However, soluble
(S1) fractions from ThyTau22 hippocampus presented relatively high content of soluble
phospho-tau (AT8-positive) and were highly toxic for microglial cells in vitro, whereas
the correspondent S1 fractions from P301S mice displayed low soluble phosphotau
levels and were not toxic for microglial cells. Therefore, not only the expression
levels but the aggregation of phospho-tau should differ between both models. In fact,
most of tau forms in the P301S mice were aggregated and, in consequence, forming
insoluble tau species.We conclude that different factors as tau mutations, accumulation,
phosphorylation, and/or aggregation could account for the distinct microglial responses
observed in these two tau models. For this reason, deciphering the molecular nature of
toxic tau species for microglial cells might be a promising therapeutic approach in order
to restore the deficient immunological protection observed in AD hippocampus.CIBERNEDJunta de Andalucía. Consejería de Economía, Innovación, Ciencia y Empleo CTS-2035Fundación Tatiana Pérez de Guzmán el BuenoMinisterio de Ciencia, Innovación y UniversidadesInstituto de Salud Carlos III. Fondo de Investigación Sanitaria. PI15/00957 PI15/00796Fondo Europeo de Desarrollo Regional PI15/00957 PI15/0079
Myelin basic protein peptide 45–89 induces the release of nitric oxide from microglial cells.
Continuous (24 h) exposure of mixed oligodendrocyte/microglial cells to peptides
45–89 derived from citrullinated C8 isoforms of myelin basic protein (MBP) induces
cell death. In contrast, MBP-C8 at the same molecular concentration is not
toxic to oligodendrocyte/microglial cells as detected by the MTT test and trypan
blue exclusion method. The loss of oligodendrocyte/microglial cells resulted in the
release of cytochrome c from mitochondria, suggesting MBP 45–89-induced
apoptosis. On the other hand, peptides 45–89 stimulated the secretion of nitric
oxide from microglial cells only via induction of iNOS. The addition of peptide
45–89 to the microglial cells led to a decrease of the level of the inhibitory protein
IkB, indicating that activation of the transcription factor NF-kB is involved in these
processes. We propose that the immunodominant peptide 45–89 induces damage of
oligodendrocytes by activation of microglial cells and subsequent generation of
nitric oxide, and that this may be the first step in the initiation of autoimmunity
Decoding damage-associated microglia in post mortem hippocampus of Alzheimer’s disease patients
The relationship between Alzheimer’s disease (AD) and neuroinflammation has become stronger since the
identification of several genetic risk factors related to microglial function. Though the role of microglial cells in the development/progression of AD is still unknown, a dysfunctional response has recently gained support. In this sense, we have reported an attenuated microglial activation associated to amyloid plaques in the hippocampus of AD patients, including a prominent degenerative process of the microglial population in the dentate gyrus, which was in contrast to the exacerbated microglial response in amyloidogenic models. This microglial degeneration could compromise their normal role of surveying the brain environment and respond to the damage. Here, we have further analyzed the phenotypic profile displayed by the damage-associated microglial cells by immunostaining and qPCR in the hippocampus of postmortem samples of AD patients (Braak V-VI) and control cases (Braak 0-II). Damage-associated microglial cells of Braak V-VI individuals were clustered around amyloid plaques and expressed Iba1, CD68, Trem2, TMEM119 and CD45high. A subset of these cells also expressed ferritin. On the contrary, these microglia down-regulated homeostatic markers, such as Cx3cr1 and P2ry12. The homeostatic and ramified microglial cells of non-demented Braak II cases were characterized by Iba1, CX3CR1, P2ry12, TMEM119 and CD45low expression. The dynamic of the microglial molecular phenotypes associated to AD pathology needs to be considered for better understand the disease complexity and, therefore, guarantee clinical success. Correcting dysregulated brain inflammatory responses might be a promising avenue to prevent/slow cognitive decline.Universidad de Málaga. Campus de excelencia Internacional-Andalucía Tech. Supported by PI18/01557 (AG) and PI18/01556 (JV) grants from ISCiii of Spain co-financed by FEDER funds from European Union
Stimulation of Na<sup>+</sup>/H<sup>+</sup> Exchanger Isoform 1 Promotes Microglial Migration
Regulation of microglial migration is not well understood. In this study, we proposed that Na+/H+ exchanger isoform 1 (NHE-1) is important in microglial migration. NHE-1 protein was co-localized with cytoskeletal protein ezrin in lamellipodia of microglia and maintained its more alkaline intracellular pH (pHi). Chemoattractant bradykinin (BK) stimulated microglial migration by increasing lamellipodial area and protrusion rate, but reducing lamellipodial persistence time. Interestingly, blocking NHE-1 activity with its potent inhibitor HOE 642 not only acidified microglia, abolished the BK-triggered dynamic changes of lamellipodia, but also reduced microglial motility and microchemotaxis in response to BK. In addition, NHE-1 activation resulted in intracellular Na+ loading as well as intracellular Ca2+ elevation mediated by stimulating reverse mode operation of Na+/Ca2+ exchange (NCXrev). Taken together, our study shows that NHE-1 protein is abundantly expressed in microglial lamellipodia and maintains alkaline pHi in response to BK stimulation. In addition, NHE-1 and NCXrev play a concerted role in BK-induced microglial migration via Na+ and Ca2+ signaling. © 2013 Shi et al
Microglial responses in the human Alzheimer’s disease frontal cortex
The continuing failure to develop an effective treatment for Alzheimer’s disease (AD) reveals the complexity for
this pathology. Increasing evidence indicates that neuroinflammation involving particularly microglial cells
contributes to AD pathogenesis. The actual view, based on the findings in APP based models, gives a
cytotoxic/proinflammatory role to activated microglia. However, we have previously reported a limited activation
and microglial degeneration in the hippocampus of AD patients in contrast with that observed in amyloidogenic
models. Here, we evaluated the microglial response in a different region of AD brains, the frontal cortex. Post
mortem tissue from controls (Braak 0-II) and AD patients (Braak V-VI) including familial cases, were obtained
from Spain Neurological Tissue Banks. Cellular (immunohistochemistry and image analysis) and molecular
(qPCR and western blots) approaches were performed. Frontal cortex of AD patients (Braak V-VI) showed
strong microglial activation similar to that observed in amyloidogenic mice. These strongly activated microglial
cells, predominantly located surrounding amyloid plaques, could drive the AD pathology and, in consequence,
could be implicated in the pathology progression. Furthermore, different microglial responses were observed
between sporadic and familial AD cases. These findings in the frontal cortex were highly in contrast to the
attenuated activation and degenerative morphology displayed by microglial cells in the hippocampus of AD
patients. Regional differences in the microglial response suggest different functional states of microglial cells in a
region-specific manner. All together, these data provide a better understanding of the immunological
mechanisms underlying AD progression and uncover new potential therapeutic targets to fight this devastating
neurodegenerative disease.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech.
Supported by PI18/01557 (AG) and PI18/01556 (JV) grants from ISCiii of Spain co-financed by FEDER funds
from European Unio
Microglial subtypes: diversity within the microglial community
Microglia are brain-resident macrophages forming the first active immune barrier in the central nervous system. They fulfill multiple functions across development and adulthood and under disease conditions. Current understanding revolves around microglia acquiring distinct phenotypes upon exposure to extrinsic cues in their environment. However, emerging evidence suggests that microglia display differences in their functions that are not exclusively driven by their milieu, rather by the unique properties these cells possess. This microglial intrinsic heterogeneity has been largely overlooked, favoring the prevailing view that microglia are a single-cell type endowed with spectacular plasticity, allowing them to acquire multiple phenotypes and thereby fulfill their numerous functions in health and disease. Here, we review the evidence that microglia might form a community of cells in which each member (or "subtype") displays intrinsic properties and performs unique functions. Distinctive features and functional implications of several microglial subtypes are considered, across contexts of health and disease. Finally, we suggest that microglial subtype categorization shall be based on function and we propose ways for studying them. Hence, we advocate that plasticity (reaction states) and diversity (subtypes) should both be considered when studying the multitasking microglia.España, Ministerio de Ciencia, Innovación y Universidades FEDER y UE RTI2018-098645-B-10
Microglial response differences between amyloidogenic transgenic models and Alzheimer’s disease patients
Aims: The continuing failure to develop an effective treatment for Alzheimer’s disease (AD) reveals the complexity for AD pathology. Increasing evidence indicates that neuroinflammation involving particularly microglial cells contributes to disease pathogenesis. Here we analyze the differences in the microglial response between APP/PS1 model and human brains.
Methods: RT-PCR, western blots, and immunostaining were performed in the hippocampus of human post mortem samples (from Braak II to Braak V-VI) and APP751SL/PS1M146L mice. In vitro studies to check the effect of S1 fractions on microglial cells were assayed.
Results: In APP based models the high Abeta accumulation triggers a prominent microglial response. On the contrary, the microglial response detected in human samples is, at least, partial or really mild. This patent difference could simple reflect the lower and probably slower Abeta production observed in human hippocampal samples, in comparison with models or could reflect the consequence of a chronic long-standing microglial activation. However, beside this differential response, we also observed a prominent microglial degenerative process in Braak V-VI samples that, indeed, could compromise their normal role of surveying the brain environment and respond to the damage. This microglial degeneration, particularly relevant at the dentate gyrus of the hippocampal formation, might be mediated by the accumulation of toxic soluble phospho-tau species.
Conclusions: These differences need to be considered when delineating animal models that better integrate the complexity of AD pathology and, therefore, guarantee clinical translation. Correcting dysregulated brain inflammatory responses might be a promising avenue to restore cognitive function.
Supported by grants FIS PI15/00796 and FIS PI15/00957 co-financed by FEDER funds from European Union, and by Junta de Andalucia Proyecto de Excelencia CTS385 2035.Financiado por FIS PI15/00796 y FIS PI15/0095, cofinanciado por los fondos FEDER de la Unión Europea, y por Junta de Andalucia Proyecto de Excelencia CTS385 2035.
Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech
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