RESUMO
A primary cilium is a membrane-bound extension from the cell surface that contains receptors for perceiving and transmitting signals that modulate cell state and activity. Primary cilia in the brain are less accessible than cilia on cultured cells or epithelial tissues because in the brain they protrude into a deep, dense network of glial and neuronal processes. Here, we investigated cilia frequency, internal structure, shape, and position in large, high-resolution transmission electron microscopy volumes of mouse primary visual cortex. Cilia extended from the cell bodies of nearly all excitatory and inhibitory neurons, astrocytes, and oligodendrocyte precursor cells (OPCs) but were absent from oligodendrocytes and microglia. Ultrastructural comparisons revealed that the base of the cilium and the microtubule organization differed between neurons and glia. Investigating cilia-proximal features revealed that many cilia were directly adjacent to synapses, suggesting that cilia are poised to encounter locally released signaling molecules. Our analysis indicated that synapse proximity is likely due to random encounters in the neuropil, with no evidence that cilia modulate synapse activity as would be expected in tetrapartite synapses. The observed cell class differences in proximity to synapses were largely due to differences in external cilia length. Many key structural features that differed between neuronal and glial cilia influenced both cilium placement and shape and, thus, exposure to processes and synapses outside the cilium. Together, the ultrastructure both within and around neuronal and glial cilia suggest differences in cilia formation and function across cell types in the brain.
Assuntos
Cílios , Animais , Cílios/ultraestrutura , Camundongos , Microscopia Eletrônica de Transmissão , Camundongos Endogâmicos C57BL , Neurônios/ultraestrutura , Neurônios/fisiologia , Córtex Visual/ultraestrutura , Córtex Visual/fisiologia , Neuroglia/ultraestrutura , Neuroglia/fisiologia , Feminino , Sinapses/ultraestrutura , Sinapses/fisiologia , MasculinoRESUMO
Herein, we propose a method to estimate the reflection coefficient of the ultrasonic wave transmitted onto an object and to display this with acoustic impedance distribution. The observation targets were glial cells, which have a rigid cytoskeleton and spread out well on a culture substrate. A reflection coefficient derived only from the cells was then obtained using a deconvolution process. In the conventional method, the deconvolution process that was performed only in the frequency domain would cause an error in the reconstructed signal, and it formed an artifact when the result was converted into the acoustic impedance image. To solve this problem, two types of deconvolution techniques were applied in either the full frequency or time-frequency domain. The results of both methods were then compared. Since the characteristic acoustic impedance is a physical property substantially equivalent to the bulk modulus, it can be considered that the internal elastic parameter is thus estimated. An analysis of the nucleus based on its position in the acoustic impedance image was then performed. The results indicated that the proposed time-frequency domain deconvolution method is able to maintain the structure of the cell, while the cell itself is free from unwanted artifacts. The nucleus was also estimated to be located toward the center of the cell, with lower acoustic impedance value than the cytoskeleton. The results of this study could contribute to establishing a method for monitoring the internal condition of cultured cells in regenerative medicine and drug discovery.
Assuntos
Microscopia Acústica/métodos , Neuroglia/ultraestrutura , Animais , Células Cultivadas , Cerebelo/citologia , Análise dos Mínimos Quadrados , Ratos , TransdutoresRESUMO
Understanding cellular architecture is essential for understanding biology. Electron microscopy (EM) uniquely visualizes cellular structures with nanometre resolution. However, traditional methods, such as thin-section EM or EM tomography, have limitations in that they visualize only a single slice or a relatively small volume of the cell, respectively. Focused ion beam-scanning electron microscopy (FIB-SEM) has demonstrated the ability to image small volumes of cellular samples with 4-nm isotropic voxels1. Owing to advances in the precision and stability of FIB milling, together with enhanced signal detection and faster SEM scanning, we have increased the volume that can be imaged with 4-nm voxels by two orders of magnitude. Here we present a volume EM atlas at such resolution comprising ten three-dimensional datasets for whole cells and tissues, including cancer cells, immune cells, mouse pancreatic islets and Drosophila neural tissues. These open access data (via OpenOrganelle2) represent the foundation of a field of high-resolution whole-cell volume EM and subsequent analyses, and we invite researchers to explore this atlas and pose questions.
Assuntos
Conjuntos de Dados como Assunto , Disseminação de Informação , Microscopia Eletrônica de Varredura , Organelas/ultraestrutura , Animais , Linhagem Celular , Células Cultivadas , Drosophila melanogaster/citologia , Drosophila melanogaster/ultraestrutura , Feminino , Complexo de Golgi/ultraestrutura , Humanos , Interfase , Ilhotas Pancreáticas/citologia , Masculino , Camundongos , Microscopia Eletrônica de Varredura/métodos , Microscopia Eletrônica de Varredura/normas , Microtúbulos/ultraestrutura , Neuroglia/ultraestrutura , Neurônios/ultraestrutura , Publicação de Acesso Aberto , Neoplasias Ovarianas/imunologia , Neoplasias Ovarianas/ultraestrutura , Ribossomos/ultraestrutura , Vesículas Sinápticas/ultraestrutura , Linfócitos T Citotóxicos/citologia , Linfócitos T Citotóxicos/imunologia , Linfócitos T Citotóxicos/ultraestruturaRESUMO
Astrocytes are complex glial cells that play many essential roles in the brain, including the fine-tuning of synaptic activity and blood flow. These roles are linked to fluctuations in intracellular Ca2+ within astrocytes. Recent advances in imaging techniques have identified localized Ca2+ transients within the fine processes of the astrocytic structure, which we term microdomain Ca2+ events. These Ca2+ transients are very diverse and occur under different conditions, including in the presence or absence of surrounding circuit activity. This complexity suggests that different signalling mechanisms mediate microdomain events which may then encode specific astrocyte functions from the modulation of synapses up to brain circuits and behaviour. Several recent studies have shown that a subset of astrocyte microdomain Ca2+ events occur rapidly following local neuronal circuit activity. In this review, we consider the physiological relevance of microdomain astrocyte Ca2+ signalling within brain circuits and outline possible pathways of extracellular Ca2+ influx through ionotropic receptors and other Ca2+ ion channels, which may contribute to astrocyte microdomain events with potentially fast dynamics.
Assuntos
Astrócitos/citologia , Sinalização do Cálcio/genética , Cálcio/metabolismo , Sinapses/genética , Astrócitos/fisiologia , Astrócitos/ultraestrutura , Circulação Sanguínea/genética , Encéfalo/metabolismo , Encéfalo/ultraestrutura , Humanos , Neuroglia/metabolismo , Neuroglia/ultraestrutura , Sinapses/ultraestruturaRESUMO
BACKGROUND: Toll-like receptor 7 (TLR7) is an innate immune receptor that detects viral single-stranded RNA and triggers the production of proinflammatory cytokines and type 1 interferons in immune cells. TLR7 agonists also modulate sensory nerve function by increasing neuronal excitability, although studies are conflicting whether sensory neurons specifically express TLR7. This uncertainty has confounded the development of a mechanistic understanding of TLR7 function in nervous tissues. METHODS: TLR7 expression was tested using in situ hybridization with species-specific RNA probes in vagal and dorsal root sensory ganglia in wild-type and TLR7 knockout (KO) mice and in guinea pigs. Since TLR7 KO mice were generated by inserting an Escherichia coli lacZ gene in exon 3 of the mouse TLR7 gene, wild-type and TLR7 (KO) mouse vagal ganglia were also labeled for lacZ. In situ labeling was compared to immunohistochemistry using TLR7 antibody probes. The effects of influenza A infection on TLR7 expression in sensory ganglia and in the spleen were also assessed. RESULTS: In situ probes detected TLR7 in the spleen and in small support cells adjacent to sensory neurons in the dorsal root and vagal ganglia in wild-type mice and guinea pigs, but not in TLR7 KO mice. TLR7 was co-expressed with the macrophage marker Iba1 and the satellite glial cell marker GFAP, but not with the neuronal marker PGP9.5, indicating that TLR7 is not expressed by sensory nerves in either vagal or dorsal root ganglia in mice or guinea pigs. In contrast, TLR7 antibodies labeled small- and medium-sized neurons in wild-type and TLR7 KO mice in a TLR7-independent manner. Influenza A infection caused significant weight loss and upregulation of TLR7 in the spleens, but not in vagal ganglia, in mice. CONCLUSION: TLR7 is expressed by macrophages and satellite glial cells, but not neurons in sensory ganglia suggesting TLR7's neuromodulatory effects are mediated indirectly via activation of neuronally-associated support cells, not through activation of neurons directly. Our data also suggest TLR7's primary role in neuronal tissues is not related to antiviral immunity.
Assuntos
Gânglios Espinais/metabolismo , Macrófagos/metabolismo , Glicoproteínas de Membrana/biossíntese , Neuroglia/metabolismo , Células Receptoras Sensoriais/metabolismo , Receptor 7 Toll-Like/biossíntese , Animais , Feminino , Gânglios Espinais/ultraestrutura , Expressão Gênica , Cobaias , Macrófagos/ultraestrutura , Glicoproteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Neuroglia/ultraestrutura , Células Receptoras Sensoriais/ultraestrutura , Receptor 7 Toll-Like/genéticaRESUMO
Diverse cell types in tissues have distinct gene expression programs, chromatin states, and nuclear architectures. To correlate such multimodal information across thousands of single cells in mouse brain tissue sections, we use integrated spatial genomics, imaging thousands of genomic loci along with RNAs and epigenetic markers simultaneously in individual cells. We reveal that cell typespecific association and scaffolding of DNA loci around nuclear bodies organize the nuclear architecture and correlate with differential expression levels in different cell types. At the submegabase level, active and inactive X chromosomes access similar domain structures in single cells despite distinct epigenetic and expression states. This work represents a major step forward in linking single-cell three-dimensional nuclear architecture, gene expression, and epigenetic modifications in a native tissue context.
Assuntos
Núcleo Celular/metabolismo , Núcleo Celular/ultraestrutura , Córtex Cerebral/citologia , Neuroglia/ultraestrutura , Neurônios/ultraestrutura , Análise de Célula Única , Animais , Córtex Cerebral/metabolismo , Cromatina/metabolismo , Cromatina/ultraestrutura , Cromossomos/metabolismo , Cromossomos/ultraestrutura , Células Endoteliais/metabolismo , Células Endoteliais/ultraestrutura , Epigênese Genética , Feminino , Genoma , Hibridização in Situ Fluorescente , Camundongos , Neuroglia/metabolismo , Neurônios/metabolismo , RNA-Seq , Transcrição Gênica , Transcriptoma , Cromossomo X/metabolismo , Cromossomo X/ultraestruturaRESUMO
BACKGROUND: In the post-GWAS era, there is an unmet need to decode the underpinning genetic etiologies of late-onset Alzheimer's disease (LOAD) and translate the associations to causation. METHODS: We conducted ATAC-seq profiling using NeuN sorted-nuclei from 40 frozen brain tissues to determine LOAD-specific changes in chromatin accessibility landscape in a cell-type specific manner. RESULTS: We identified 211 LOAD-specific differential chromatin accessibility sites in neuronal-nuclei, four of which overlapped with LOAD-GWAS regions (±100 kb of SNP). While the non-neuronal nuclei did not show LOAD-specific differences, stratification by sex identified 842 LOAD-specific chromatin accessibility sites in females. Seven of these sex-dependent sites in the non-neuronal samples overlapped LOAD-GWAS regions including APOE. LOAD loci were functionally validated using single-nuclei RNA-seq datasets. CONCLUSIONS: Using brain sorted-nuclei enabled the identification of sex-dependent cell type-specific LOAD alterations in chromatin structure. These findings enhance the interpretation of LOAD-GWAS discoveries, provide potential pathomechanisms, and suggest novel LOAD-loci.
Assuntos
Doença de Alzheimer/genética , Cromatina/ultraestrutura , Neuroglia/ultraestrutura , Caracteres Sexuais , Idoso , Idoso de 80 Anos ou mais , Sequência de Bases , Sítios de Ligação , Fracionamento Celular/métodos , Núcleo Celular/ultraestrutura , Cromatina/genética , Conjuntos de Dados como Assunto , Feminino , Citometria de Fluxo , Expressão Gênica , Biblioteca Gênica , Estudo de Associação Genômica Ampla , Humanos , Masculino , Pessoa de Meia-Idade , Neurônios/ultraestrutura , Análise de Célula Única , Lobo Temporal/ultraestrutura , Fatores de Transcrição/metabolismoRESUMO
The axon initial segment (AIS) is structurally and functionally distinct from other regions of the axon, yet alterations in the milieu of the AIS after brain injury have not been well characterized. In this study, we have examined extracellular and intracellular changes in the AIS after hypoglossal nerve injury. Microglial adhesions to the AIS were rarely observed in healthy controls, whereas microglial adhesions to the AIS became apparent in the axonal injury model. Regarding intra-AIS morphology, we focused on mitochondria because mitochondrial flow into the injured axon appears critical for axonal regeneration. To visualize mitochondria specifically in injured axons, we used Atf3:BAC transgenic mice whose mitochondria were labeled with GFP in response to nerve injury. These mice clearly showed mitochondrial localization in the AIS after nerve injury. To precisely confirm the light microscopic observations, we performed three-dimensional ultrastructural analysis using focused ion beam/scanning electron microscopy (FIB/SEM). Although the healthy AIS was not surrounded by microglia, tight microglial adhesions with thick processes adhering to the AIS were observed after injury. FIB/SEM simultaneously allowed the observation of mitochondrial localization in the AIS. In the AIS of non-injured neurons, few mitochondria were observed, whereas mitochondria were abundantly localized in the cell body, axon hillock, and axon. Intriguingly, in the injured AIS, numerous mitochondria were observed throughout the AIS. Taken together, axonal injury changes the extracellular glial environment surrounding the AIS and intracellular mitochondrial localization in the AIS. These changes would be crucial responses, perhaps for injured neurons to regenerate after axonal injury.
Assuntos
Segmento Inicial do Axônio/fisiologia , Axônios/fisiologia , Espaço Extracelular/fisiologia , Mitocôndrias/fisiologia , Neuroglia/fisiologia , Fator 3 Ativador da Transcrição/genética , Animais , Segmento Inicial do Axônio/ultraestrutura , Axônios/ultraestrutura , Adesão Celular , Feminino , Humanos , Imageamento Tridimensional , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Mitocôndrias/ultraestrutura , Compressão Nervosa , Neuroglia/ultraestruturaRESUMO
BACKGROUND & AIMS: Neuroinflammation in the gut is associated with many gastrointestinal (GI) diseases, including inflammatory bowel disease. In the brain, neuroinflammatory conditions are associated with blood-brain barrier (BBB) disruption and subsequent neuronal injury. We sought to determine whether the enteric nervous system is similarly protected by a physical barrier and whether that barrier is disrupted in colitis. METHODS: Confocal and electron microscopy were used to characterize myenteric plexus structure, and FITC-dextran assays were used to assess for presence of a barrier. Colitis was induced with dextran sulfate sodium, with co-administration of liposome-encapsulated clodronate to deplete macrophages. RESULTS: We identified a blood-myenteric barrier (BMB) consisting of extracellular matrix proteins (agrin and collagen-4) and glial end-feet, reminiscent of the BBB, surrounded by a collagen-rich periganglionic space. The BMB is impermeable to the passive movement of 4 kDa FITC-dextran particles. A population of macrophages is present within enteric ganglia (intraganglionic macrophages [IGMs]) and exhibits a distinct morphology from muscularis macrophages, with extensive cytoplasmic vacuolization and mitochondrial swelling but without signs of apoptosis. IGMs can penetrate the BMB in physiological conditions and establish direct contact with neurons and glia. Dextran sulfate sodium-induced colitis leads to BMB disruption, loss of its barrier integrity, and increased numbers of IGMs in a macrophage-dependent process. CONCLUSIONS: In intestinal inflammation, macrophage-mediated degradation of the BMB disrupts its physiological barrier function, eliminates the separation of the intra- and extra-ganglionic compartments, and allows inflammatory stimuli to access the myenteric plexus. This suggests a potential mechanism for the onset of neuroinflammation in colitis and other GI pathologies with acquired enteric neuronal dysfunction.
Assuntos
Colite/etiologia , Colite/metabolismo , Macrófagos/imunologia , Macrófagos/metabolismo , Plexo Mientérico/citologia , Plexo Mientérico/metabolismo , Animais , Biomarcadores , Colite/patologia , Modelos Animais de Doenças , Suscetibilidade a Doenças , Sistema Nervoso Entérico/imunologia , Sistema Nervoso Entérico/metabolismo , Matriz Extracelular , Imunofluorescência , Imuno-Histoquímica , Imunofenotipagem , Camundongos , Plexo Mientérico/ultraestrutura , Neuroglia/metabolismo , Neuroglia/ultraestrutura , Doenças Neuroinflamatórias/etiologia , Doenças Neuroinflamatórias/metabolismo , Doenças Neuroinflamatórias/patologia , Infiltração de NeutrófilosRESUMO
Cellular heterogeneity in the human brain obscures the identification of robust cellular regulatory networks, which is necessary to understand the function of non-coding elements and the impact of non-coding genetic variation. Here we integrate genome-wide chromosome conformation data from purified neurons and glia with transcriptomic and enhancer profiles, to characterize the gene regulatory landscape of two major cell classes in the human brain. We then leverage cell-type-specific regulatory landscapes to gain insight into the cellular etiology of several brain disorders. We find that Alzheimer's disease (AD)-associated epigenetic dysregulation is linked to neurons and oligodendrocytes, whereas genetic risk factors for AD highlighted microglia, suggesting that different cell types may contribute to disease risk, via different mechanisms. Moreover, integration of glutamatergic and GABAergic regulatory maps with genetic risk factors for schizophrenia (SCZ) and bipolar disorder (BD) identifies shared (parvalbumin-expressing interneurons) and distinct cellular etiologies (upper layer neurons for BD, and deeper layer projection neurons for SCZ). Collectively, these findings shed new light on cell-type-specific gene regulatory networks in brain disorders.
Assuntos
Doença de Alzheimer/genética , Transtorno Bipolar/genética , Cromatina/ultraestrutura , Esquizofrenia/genética , Acetilação , Doença de Alzheimer/patologia , Transtorno Bipolar/patologia , Cromatina/química , Cromatina/genética , Cromatina/metabolismo , Sequenciamento de Cromatina por Imunoprecipitação , Elementos Facilitadores Genéticos , Epigênese Genética , Neurônios GABAérgicos/metabolismo , Regulação da Expressão Gênica , Estudo de Associação Genômica Ampla , Histonas/metabolismo , Humanos , Lisina/metabolismo , Neuroglia/patologia , Neuroglia/ultraestrutura , Neurônios/patologia , Neurônios/ultraestrutura , Regiões Promotoras Genéticas , Esquizofrenia/patologiaRESUMO
Our brains consist of 80% water, which is continuously shifted between different compartments and cell types during physiological and pathophysiological processes. Disturbances in brain water homeostasis occur with pathologies such as brain oedema and hydrocephalus, in which fluid accumulation leads to elevated intracranial pressure. Targeted pharmacological treatments do not exist for these conditions owing to our incomplete understanding of the molecular mechanisms governing brain water transport. Historically, the transmembrane movement of brain water was assumed to occur as passive movement of water along the osmotic gradient, greatly accelerated by water channels termed aquaporins. Although aquaporins govern the majority of fluid handling in the kidney, they do not suffice to explain the overall brain water movement: either they are not present in the membranes across which water flows or they appear not to be required for the observed flow of water. Notably, brain fluid can be secreted against an osmotic gradient, suggesting that conventional osmotic water flow may not describe all transmembrane fluid transport in the brain. The cotransport of water is an unconventional molecular mechanism that is introduced in this Review as a missing link to bridge the gap in our understanding of cellular and barrier brain water transport.
Assuntos
Encéfalo/metabolismo , Água/metabolismo , Animais , Aquaporinas/metabolismo , Água Corporal/metabolismo , Proteínas de Transporte/metabolismo , Membrana Celular/metabolismo , Tamanho Celular , Líquido Cefalorraquidiano/metabolismo , Endotélio Vascular/metabolismo , Líquido Extracelular/metabolismo , Sistema Glinfático/fisiologia , Humanos , Líquido Intracelular/metabolismo , Transporte de Íons , Proteínas do Tecido Nervoso/metabolismo , Neuroglia/metabolismo , Neuroglia/ultraestrutura , Neurônios/metabolismo , Neurônios/ultraestrutura , Osmose , Potássio/metabolismo , ATPase Trocadora de Sódio-Potássio/metabolismo , Espaço SubaracnóideoRESUMO
We describe here phase-separated subnuclear organelles in the nematode Caenorhabditis elegans, which we term NUN (NUclear Nervous system-specific) bodies. Unlike other previously described subnuclear organelles, NUN bodies are highly cell type specific. In fully mature animals, 4-10 NUN bodies are observed exclusively in the nucleus of neuronal, glial and neuron-like cells, but not in other somatic cell types. Based on co-localization and genetic loss of function studies, NUN bodies are not related to other previously described subnuclear organelles, such as nucleoli, splicing speckles, paraspeckles, Polycomb bodies, promyelocytic leukemia bodies, gems, stress-induced nuclear bodies, or clastosomes. NUN bodies form immediately after cell cycle exit, before other signs of overt neuronal differentiation and are unaffected by the genetic elimination of transcription factors that control many other aspects of neuronal identity. In one unusual neuron class, the canal-associated neurons, NUN bodies remodel during larval development, and this remodeling depends on the Prd-type homeobox gene ceh-10. In conclusion, we have characterized here a novel subnuclear organelle whose cell type specificity poses the intriguing question of what biochemical process in the nucleus makes all nervous system-associated cells different from cells outside the nervous system.
Assuntos
Espaço Intranuclear/ultraestrutura , Neurônios/ultraestrutura , Animais , Caenorhabditis elegans , Proteínas de Caenorhabditis elegans/genética , Proteínas de Caenorhabditis elegans/metabolismo , Ciclo Celular , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Espaço Intranuclear/metabolismo , Neuroglia/ultraestruturaRESUMO
The Locus Coeruleus (LC) is a pontine nucleus involved in many physiological processes, including the control of the sleep/wake cycle (SWC). At cellular level, the LC displays a high density of opioid receptors whose activation decreases the activity of LC noradrenergic neurons. Also, microinjections of morphine administered locally in the LC of the cat produce sleep associated with synchronized brain activity in the electroencephalogram (EEG). Even though much of the research on sleep has been done in the cat, the subcellular location of opioid receptors in the LC and their relationship with LC noradrenergic neurons is not known yet in this species. Therefore, we conducted a study to describe the ultrastructural localization of mu-opioid receptors (MOR), delta-opioid receptors (DOR) and tyrosine hydroxylase (TH) in the cat LC using high resolution electron microscopy double-immunocytochemical detection. MOR and DOR were localized mainly in dendrites (45% and 46% of the total number of profiles respectively), many of which were noradrenergic (35% and 53% for MOR and DOR, respectively). TH immunoreactivity was more frequent in dendrites (65% of the total number of profiles), which mostly also expressed opioid receptors (58% and 73% for MOR and DOR, respectively). Because the distribution of MORs and DORs are similar, it is possible that a substantial sub-population of neurons co-express both receptors, which may facilitate the formation of MOR-DOR heterodimers. Moreover, we found differences in the cat subcellular DOR distribution compared with the rat. This opens the possibility to the existence of diverse mechanisms for opioid modulation of LC activity.
Assuntos
Neurônios Adrenérgicos/ultraestrutura , Dendritos/ultraestrutura , Locus Cerúleo/ultraestrutura , Neuroglia/ultraestrutura , Receptores Opioides delta/ultraestrutura , Receptores Opioides mu/ultraestrutura , Neurônios Adrenérgicos/metabolismo , Animais , Gatos , Dendritos/metabolismo , Locus Cerúleo/metabolismo , Neuroglia/metabolismo , Receptores Opioides delta/metabolismo , Receptores Opioides mu/metabolismoRESUMO
Airway pollution can affect the central nervous system, but whether this causes glial activation and inflammation in the nucleus of solitary tract (NTS) remains unclear. We used a rat model with exposure to diesel exhaust particulate matter (DEP) at 200 µg/m3 (low exposure) and 1000 µg/m3 (high exposure) for 14 days. Activation of microglia and astrocytes in the NTS was assessed using Iba-1 and glial fibrillary acidic protein (GFAP) staining. The expression of neurotrophic factors including brain-derived neurotrophic factor (BDNF), glial-derived neurotrophic factor (GDNF), and nerve growth factor (NGF) in the NTS were evaluated by immunofluorescence. Changes in the intracellular structure of NTS neurons were observed via electron microscopy. Inflammatory cytokines and oxidant stress levels in the medulla were also measured. Exposure to DEP can cause NTS inflammation as well as airway inflammation, especially in the H-exposure group. We showed that the numbers of microglia and astrocytes in the NTS, as well as NGF expression in the NTS, were significantly higher in both exposure groups than in controls, but BDNF or GDNF expression was not detected. Exposure to DEP induced ultrastructural changes in NTS neurons as reflected by endoplasmic reticulum dilation, ribosomal loss, mitochondrial vacuolization, and a sparse myelin sheath. Medulla inflammation and an imbalance of oxidants and antioxidants also resulted from exposure to DEP. The H-exposure group showed an imbalance of oxidants and antioxidants with decreased levels of SOD and GSH and increased levels of MDA and ROS compared to the control group (both p < 0.01) in the medulla. Inflammatory cytokines (IL-1ß, IL-6, and TNF-α) were also significantly increased in the H-exposure group. Fourteen days of exposure to DEP can affect the NTS neurons in rat. Glial activation and inflammation may play important roles in the response of the NTS to DEP.
Assuntos
Poluentes Atmosféricos/toxicidade , Encéfalo/efeitos dos fármacos , Pulmão/efeitos dos fármacos , Neuroglia/efeitos dos fármacos , Emissões de Veículos/toxicidade , Administração por Inalação , Animais , Encéfalo/patologia , Encéfalo/ultraestrutura , Inflamação/etiologia , Inflamação/patologia , Pulmão/patologia , Masculino , Microscopia Eletrônica , Neuroglia/patologia , Neuroglia/ultraestrutura , Ratos Sprague-DawleyRESUMO
Studying the spatiotemporal dynamic changes of various cells following spinal cord injury (SCI) is of great significance for understanding the pathological processes of SCI. Changes in the characteristics of Sox9-positive cells, which are widely present in the spinal cord, have rarely been studied following SCI. We found that Sox9-positive cells were widely distributed in the central canal and parenchyma of the uninjured adult spinal cord, with the greatest distribution in the central spinal cord and relatively few cells in the dorsal and ventral sides. Ranging between 14.20% ± 1.61% and 15.60% ± 0.36% of total cells in the spinal cord, almost all Sox9-positive cells were in a quiescent state. However, Sox9-positive cells activated following SCI exhibited different characteristics according to their distance from the lesion area. In the reactive region, Sox9-positive cells highly expressed nestin and exhibited a single-branching structure, whereas in the non-reactive region, cells showed low nestin expression and a multi-branching structure. In response to SCI, a large number of Sox9-positive cells in the spinal cord parenchyma proliferated to participate in the formation of glial scars, whereas Sox9-positive cells in the central canal located near the lesion site accumulated at its broken ends through proliferation. Finally, we found that approximately 6.30% ± 0.35% of Sox9-positive cells differentiated into oligodendrocytes within two weeks after SCI. By examining the spatiotemporal dynamic changes, proliferation and differentiation characteristics of Sox9-positive cells after SCI, our findings provide a theoretical basis for understanding the pathological process of SCI.
Assuntos
Fatores de Transcrição SOX9/genética , Traumatismos da Medula Espinal/genética , Traumatismos da Medula Espinal/patologia , Animais , Bromodesoxiuridina/farmacologia , Diferenciação Celular , Proliferação de Células , Antagonistas de Estrogênios/farmacologia , Proteína Glial Fibrilar Ácida/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Nestina/metabolismo , Neuroglia/patologia , Neuroglia/ultraestrutura , Oligodendroglia/patologia , Oligodendroglia/ultraestrutura , Medula Espinal/patologia , Medula Espinal/ultraestrutura , Tamoxifeno/farmacologiaRESUMO
In neuroanatomy textbooks on humans, the posterior median septum is commonly depicted along the midline of the posterior column of the spinal cord. For intramedullary spinal cord tumors, the standard surgical treatment is posterior midline myelotomy. However, its anatomical basis is still unclear. Therefore, in this study we focused on the ultrastructural characterization of the median structure of the posterior column in an adult rat. In the median part of the fasciculi gracilis, a fine lineal tissue continued from the posterior median sulcus to the 3/4th depth of the fasciculi. At higher magnification, this fine lineal tissue consisted of bundles of astrocytes, which are often disrupted and eventually disappeared. At the junction of the ventral part of the fasciculi cuneatus and the gray commissure, short lineal figures of glial tissues extended dorsally. These lineal figures of glial tissues were morphologically similar to other lineal figures of glial tissues found in the posterior column; bundles of astrocytes extending along the axons that entered the gray commissure and the perivascular lineal figures of glial tissues. In conclusion, this study revealed that the posterior median septum is composed of very fine lineal figures of glial tissues that are often disrupted and eventually disappear. We consider these basic structures to be similar in humans. Therefore, during posterior midline myelotomy, accurately separating along the posterior median septum in the posterior column is extremely difficult.
Assuntos
Neuroglia/ultraestrutura , Pia-Máter/ultraestrutura , Medula Espinal/ultraestrutura , Animais , Axônios/ultraestrutura , Masculino , Microscopia Eletrônica , Ratos , Ratos WistarRESUMO
The current study sought to characterize the pro-survival effects of erythropoietin (EPO) in a toxicant model of Parkinson's disease (PD). EPO treatment induced time-dependent elevations of antioxidant glutathione peroxidase (GPx) and anti-apoptotic factors (pAkt and pBad/Bad) within the striatum and substantia nigra pars compacta (SNc). Intriguingly, our results indicated a region- and lesion size- dependence of pro-survival effects of EPO. Indeed, intra-striatal (but not intra-nigral) infusion of EPO was effective at preventing dopaminergic terminal degeneration and sSNc neuronal loss induced by two different doses of 6-OHDA. These neuroprotective consequences were paralleled by a diminution of microglial morphological changes, along with enhanced motor functioning seen through a reduction in apomorphine-induced rotational behaviour. Finally, in the context of the 6-OHDA lesion, EPO again induced anti-apoptotic (Bcl-2) and antioxidant (GPx) factors within the striatum. Taken together, these results raise the possibility of EPO's potential use as an adjuvant therapy in the treatment of PD, or at least, suggest possible brain-region specific targets for the protective effects of EPO.
Assuntos
Corpo Estriado/efeitos dos fármacos , Eritropoetina/uso terapêutico , Fármacos Neuroprotetores/uso terapêutico , Estresse Oxidativo/efeitos dos fármacos , Transtornos Parkinsonianos/tratamento farmacológico , Animais , Apomorfina/farmacologia , Encéfalo/efeitos dos fármacos , Encéfalo/patologia , Forma Celular/efeitos dos fármacos , Corpo Estriado/patologia , Dopamina/metabolismo , Eritropoetina/farmacologia , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Microglia/efeitos dos fármacos , Microglia/ultraestrutura , Neuroglia/efeitos dos fármacos , Neuroglia/ultraestrutura , Fármacos Neuroprotetores/farmacologia , Oxidopamina/toxicidade , Transtornos Parkinsonianos/induzido quimicamente , Transtornos Parkinsonianos/metabolismo , Projetos Piloto , Proteínas Recombinantes/farmacologia , Proteínas Recombinantes/uso terapêutico , Método Simples-Cego , Gravação em VídeoRESUMO
Electron microscopy revealed that glial cells in the posterior sub-esophageal mass of the brain in Sepia officinalis had a well-developed rough endoplasmic reticulum formed by long coverslips with rectilinear or curvilinear arrangements. The coverslips appeared dilated and have a large amount of adhered polysomes. Vesicular lamellae coexisted with the elongated lamellae of RER and dictyosomes of Golgi apparatus. Endocytosis was evidenced through the pale vesicles which were appeared next to the apical border of microvilli in some glial cells. Sub-cellular features of endocytosis, predominantly the fluid phase, were observed in the apical glial cell cytoplasm. Glial cells were related to phagocytosis of apoptotic neurons, endocytosis, pinocytosis and adsorption. These functions were proposed based on their ultrastructure characteristics and a significant number of vesicles with different shapes (oval to polygonal), sizes 0.052-0.67 µm and contents. Glycogen, MPS and lipid were detected in the glial cells. Alkaline phosphatase was not observed, while an activity of acid phosphatase was bound to lysosomes. ATPases were present in the glial cells along the lateral and basal plasma lemma as well as on the membranes of cell organelles. Unspecific esterase was clearly recognizable by electron microscopy. The monoamine and cytochrome oxidase activities were demonstrated, while the succinate dehydrogenase showed a weak enzyme activity.
Assuntos
Encéfalo/citologia , Neuroglia/química , Neuroglia/ultraestrutura , Sepia/citologia , Animais , Endocitose , Retículo Endoplasmático/ultraestrutura , Complexo de Golgi/ultraestrutura , Microscopia Eletrônica , PolirribossomosRESUMO
This study aims to investigate the fine structure of the different cell types in the central brain of Eledone cirrhosa; the organelles in the neurons and the glial cells; the glial hemolymph-brain barrier; the neuro-secretions and the relationships between glial and nerve cells. The brain is surrounded by a non-cellular neurilemma followed by a single layer of perilemmal cells. Ependymal cells, highly prismatic glial cells, astrocytes, oligodendrocytes and epithelial processes were observed. The perikarya of the neurons are filled with slightly oval nuclei with heterochromatin, a strongly tortuous ER, numerous mitochondria and Golgi apparatus with two types of vesicles. In the cellular cortex, glial cells are much less numerous than the neurons and they are located preferably at the border between perikarya and neuropil. Furthermore, they send many branching shoots between the surrounding neuron perikarya and the axons. The glial cytoplasmic matrix appears more electrodense than that of the neurons. Only few ribosomes are attached to the membranes of the ER; the vast majorities are free. In the perikarya of the glial cells, mitochondria, multi-vesicular bodies, various vacuoles and vesicles are present. The essential elements of the hemolymph-brain barrier are the endothelial cells with their tight junctions. The cytoplasm contains various vesicles and mitochondria. However, two other cell types are present, the pericytes and the astrocytes, which are of great importance for the function of the hemolymph-brain barrier. The cell-cell interactions between endothelial cells, pericytes and astrocytes are as close as no other cells.
Assuntos
Encéfalo/citologia , Células Endoteliais/citologia , Neuroglia/citologia , Neurônios/citologia , Octopodiformes/citologia , Animais , Axônios/ultraestrutura , Encéfalo/ultraestrutura , Células Endoteliais/ultraestrutura , Neuroglia/ultraestrutura , Neurônios/ultraestruturaRESUMO
The transected lumbar spinal cord of lizards was studied for its ability to recover after paralysis. At 34 days post-lesion about 50% of lizards were capable of walking with a limited coordination, likely due to the regeneration of few connecting axons crossing the transection site of the spinal cord. This region, indicated as "bridge", contains glial cells among which oligodendrocytes and their elongation that are immunolabeled for NOGO-A. A main reactive protein band occurs at 100-110 kDa but a weaker band is also observed around 240 kDa, suggesting fragmentation of the native protein due to extraction or to physiological processing of the original protein. Most of the cytoplasmic immunolabeling observed in oligodendrocytes is associated with vesicles of the endoplasmic reticulum. Also, the nucleus is labeled in some oligodendrocytes that are myelinating sparse axons observed within the bridge at 22-34 days post-transection. This suggests that axonal regeneration is present within the bridge region. Immunolabeling for NOGO-A shows that the protein is also present in numerous reactive neurons, in particular motor-neurons localized in the proximal stump of the transected spinal cord. Ultrastructural immunolocalization suggests that NOGO is synthesized in the ribosomes of these neurons and becomes associated with the cisternae of the endoplasmic reticulum, probably following a secretory pathway addressed toward the axon. The present observations suggest that, like for the regenerating spinal cord of fish and amphibians, also in lizard NOGO-A is present in reactive neurons and appears associated to axonal regeneration and myelination.