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Isolation microglia

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https://www.readbyqxmd.com/read/28546318/an-environment-dependent-transcriptional-network-specifies-human-microglia-identity
#1
David Gosselin, Dylan Skola, Nicole G Coufal, Inge R Holtman, Johannes C M Schlachetzki, Eniko Sajti, Baptiste N Jaeger, Carolyn O'Connor, Conor Fitzpatrick, Martina P Pasillas, Monique Pena, Amy Adair, David G Gonda, Michael L Levy, Richard M Ransohoff, Fred H Gage, Christopher K Glass
Microglia play essential roles in central nervous system (CNS) homeostasis and influence diverse aspects of neuronal function. However, the transcriptional mechanisms that specify human microglia phenotypes are largely unknown. We examined the transcriptomes and epigenetic landscapes of human microglia isolated from surgically resected brain tissue ex vivo and following transition to an in vitro environment. Transfer to a tissue culture environment results in rapid and extensive downregulation of microglia-specific genes that are induced in primitive mouse macrophages following migration into the fetal brain...
May 25, 2017: Science
https://www.readbyqxmd.com/read/28534246/microglia-housekeeper-of-the-central-nervous-system
#2
REVIEW
John Alimamy Kabba, Yazhou Xu, Handson Christian, Wenchen Ruan, Kitchen Chenai, Yun Xiang, Luyong Zhang, Juan M Saavedra, Tao Pang
Microglia, of myeloid origin, play fundamental roles in the control of immune responses and the maintenance of central nervous system homeostasis. These cells, just like peripheral macrophages, may be activated into M1 pro-inflammatory or M2 anti-inflammatory phenotypes by appropriate stimuli. Microglia do not respond in isolation, but form part of complex networks of cells influencing each other. This review addresses the complex interaction of microglia with each cell type in the brain: neurons, astrocytes, cerebrovascular endothelial cells, and oligodendrocytes...
May 22, 2017: Cellular and Molecular Neurobiology
https://www.readbyqxmd.com/read/28521131/diverse-requirements-for-microglial-survival-specification-and-function-revealed-by-defined-medium-cultures
#3
Christopher J Bohlen, F Chris Bennett, Andrew F Tucker, Hannah Y Collins, Sara B Mulinyawe, Ben A Barres
Microglia, the resident macrophages of the CNS, engage in various CNS-specific functions that are critical for development and health. To better study microglia and the properties that distinguish them from other tissue macrophage populations, we have optimized serum-free culture conditions to permit robust survival of highly ramified adult microglia under defined-medium conditions. We find that astrocyte-derived factors prevent microglial death ex vivo and that this activity results from three primary components, CSF-1/IL-34, TGF-β2, and cholesterol...
May 17, 2017: Neuron
https://www.readbyqxmd.com/read/28508369/spike-in-silac-approach-for-proteomic-analysis-of-ex-vivo-microglia
#4
Joao Paulo Costa Pinho, Harris Bell-Temin, Bin Liu, Stanley M Stevens
Stable isotope labeling by amino acids in cell culture (SILAC) is a versatile mass spectrometry-based proteomic approach that can achieve accurate relative protein quantitation on a global scale. In this approach, proteins are labeled while being synthesized by the cell due to the presence of certain amino acids exclusively as heavier mass analogs than their regular (light) counterparts. This differential labeling allows for the identification of heavy and light forms of each peptide corresponding to two or more different experimental groups upon mass spectrometric analysis, the intensities of which reflect their abundance in the sample analyzed...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28495139/high-yield-primary-microglial-cultures-using-granulocyte-macrophage-colony-stimulating-factor-from-embryonic-murine-cerebral-cortical-tissue
#5
Adam C Yu, Sarah E Neil, Jacqueline A Quandt
BACKGROUND: Microglia play vital roles in neurotrophic support and modulating immune or inflammatory responses to pathogens or damage/stressors during disease. This study describes the ability to establish large numbers of microglia from embryonic tissues with the addition of granulocyte-macrophage stimulating factor (GM-CSF) and characterizes their similarities to adult microglia examined ex vivo as well as their responses to inflammatory mediators. METHOD: Microglia were seeded from a primary embryonic mixed cortical suspension with the addition of GM-CSF...
June 15, 2017: Journal of Neuroimmunology
https://www.readbyqxmd.com/read/28494583/isolation-identification-and-biological-evaluation-of-phenolic-compounds-from-the-traditional-north-american-confectionery-maple-sugar
#6
Yongqiang Liu, Kenneth Rose, Nicholas Dasilva, Shelby Johnson, Navindra P Seeram
Maple sap, collected from the sugar maple (Acer saccharum) tree, is boiled to produce the popular plant-derived sweetener, maple syrup, which can then be further evaporated to yield the traditional North American confectionery, maple sugar. While maple sap and maple syrup have been previously studied, the phytochemical constituents of maple sugar are unknown. Herein, thirty phenolic compounds, 1-30, primarily lignans, were isolated and identified (by HRESIMS and NMR) from maple sugar. The isolates included the phenylpropanoid-based lignan tetramers, (erythro,erythro)-4'',4'''-dihydroxy-3,3',3'',3''',5,5'-hexamethoxy-7,9';7',9-diepoxy-4,8'';4',8'''-bisoxy-8,8'-dineolignan-7'',7''',9'',9'''-tetraol, 29, and (threo,erythro)-4'',4'''-dihydroxy-3,3',3'',3''',5,5'-hexamethoxy-7,9';7',9-diepoxy-4,8'';4',8'''-bisoxy-8,8'-dineolignan-7'',7''',9'',9'''-tetraol, 30, neither of which have been identified from maple sap or maple syrup before...
May 12, 2017: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/28486838/brain-microglial-cells-are-highly-susceptible-to-hiv-1-infection-and-spread
#7
Jennifer J Cenker, Ryan D Stultz, David McDonald
Macrophages are a target of HIV-1 infection and may serve as an important reservoir of the virus in the body, particularly after depletion of CD4+ T cells in HIV/AIDS. Recently, sterile alpha motif and histidine/aspartic acid domain-containing protein 1 (SAMHD1) was identified as the major restriction factor of HIV-1 infection in myeloid cells. SAMHD1 is targeted for proteolytic degradation by Vpx, a viral protein encoded by HIV-2 and many SIVs but not HIV-1. In this study, we assessed SAMHD1 restriction in in vitro differentiated macrophages and in freshly isolated macrophages from the lungs, abdomen, and brain...
May 9, 2017: AIDS Research and Human Retroviruses
https://www.readbyqxmd.com/read/28482909/characteristics-of-primary-rat-microglia-isolated-from-mixed-cultures-using-two-different-methods
#8
Li Lin, Rakhi Desai, Xiaoying Wang, Eng H Lo, Changhong Xing
BACKGROUND: Microglial cultures comprise a critically important model system for investigating inflammatory mechanisms in almost all CNS disorders. Mild trypsinization and shaking are the two most commonly used methods to isolate primary microglia from mixed glial cultures. In this study, we characterized and compared microglia obtained using these two methods. METHODS: Primary rat microglia cultures were prepared from cerebral cortices of 1-2-day-old neonatal Sprague-Dawley rats...
May 8, 2017: Journal of Neuroinflammation
https://www.readbyqxmd.com/read/28474151/alismol-a-sesquiterpenoid-isolated-from-vladimiria-souliei-suppresses-proinflammatory-mediators-in-lipopolysaccharide-stimulated-microglia
#9
Dongdong Shi, Xiaoqiang Song, Yuanqiang Guo, Jing Xu, Yu Liu, Jingling Zhang, Chun-Ai Cui, Da-Qing Jin
Microglia activation plays an important role in the pathogenesis of various neurodegenerative diseases by producing neurotoxic factors, such as proinflammatory cytokines and nitric oxide (NO); therefore, suppression of microglia activation is a potential therapeutic approach against these diseases. Previous study showed that alismol, a sesquiterpenoid isolated from the roots of Vladimiria souliei inhibits interferon-γ-induced NO production in murine macrophage RAW264.7 cells. In the present study, we found that alismol reduced NO and prostaglandin E2 (PGE2) levels and suppressed the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-stimulated primary and cultured microglia...
May 4, 2017: Journal of Molecular Neuroscience: MN
https://www.readbyqxmd.com/read/28458018/anti-inflammatory-effects-of-novel-polygonum-multiflorum-compound-via-inhibiting-nf-%C3%AE%C2%BAb-mapk-and-upregulating-the-nrf2-pathways-in-lps-stimulated-microglia
#10
Sun Young Park, Mei Ling Jin, Nam Jun Kang, Geuntae Park, Young-Whan Choi
The incorporation of Polygonum multiflorum into the diet can result in anti-aging effects owing to its wide range of biological and pharmaceutical properties. We investigated the anti-neuroinflammatory properties of CRPE56IGIH isolated from P. multiflorum by focusing on its role in the induction of phase II antioxidant enzymes and the modulation of upstream signaling pathways. In microglia, CRPE56IGIH significantly inhibited lipopolysaccharide (LPS)-stimulated nitric oxide and prostaglandin E2 production with nonspecific cytotoxicity...
April 27, 2017: Neuroscience Letters
https://www.readbyqxmd.com/read/28447995/rapid-and-refined-cd11b-magnetic-isolation-of-primary-microglia-with-enhanced-purity-and-versatility
#11
Souvarish Sarkar, Emir Malovic, Brandon Plante, Gary Zenitsky, Huajun Jin, Vellareddy Anantharam, Arthi Kanthasamy, Anumantha G Kanthasamy
Microglia are the primary responders to central nervous system insults; however, much remains unknown about their role in regulating neuroinflammation. Microglia are mesodermal cells that function similarly to macrophages in surveying inflammatory stress. The classical (M1-type) and alternative (M2-type) activations of macrophages have also been extended to microglia in an effort to better understand the underlying interplay these phenotypes have in neuroinflammatory conditions such as Parkinson's, Alzheimer's, and Huntington's Diseases...
April 13, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28446241/polyunsaturated-fatty-acid-receptors-gpr40-and-gpr120-are-expressed-in-the-hypothalamus-and-control-energy-homeostasis-and-inflammation
#12
Nathalia R V Dragano, Carina Solon, Albina F Ramalho, Rodrigo F de Moura, Daniela S Razolli, Elisabeth Christiansen, Carlos Azevedo, Trond Ulven, Licio A Velloso
BACKGROUND: The consumption of large amounts of dietary fats is one of the most important environmental factors contributing to the development of obesity and metabolic disorders. GPR120 and GPR40 are polyunsaturated fatty acid receptors that exert a number of systemic effects that are beneficial for metabolic and inflammatory diseases. Here, we evaluate the expression and potential role of hypothalamic GPR120 and GPR40 as targets for the treatment of obesity. METHODS: Male Swiss (6-weeks old), were fed with a high fat diet (HFD, 60% of kcal from fat) for 4 weeks...
April 26, 2017: Journal of Neuroinflammation
https://www.readbyqxmd.com/read/28437540/characterization-of-gene-expression-phenotype-in-amyotrophic-lateral-sclerosis-monocytes
#13
Weihua Zhao, David R Beers, Kristopher G Hooten, Douglas H Sieglaff, Aijun Zhang, Shanker Kalyana-Sundaram, Christopher M Traini, Wendy S Halsey, Ashley M Hughes, Ganesh M Sathe, George P Livi, Guo-Huang Fan, Stanley H Appel
Importance: Amyotrophic lateral sclerosis (ALS) is a common adult-onset neurodegenerative disease characterized by selective loss of upper and lower motor neurons. Patients with ALS have persistent peripheral and central inflammatory responses including abnormally functioning T cells and activated microglia. However, much less is known about the inflammatory gene profile of circulating innate immune monocytes in these patients. Objective: To characterize the transcriptomics of peripheral monocytes in patients with ALS...
April 24, 2017: JAMA Neurology
https://www.readbyqxmd.com/read/28434692/immune-hyperreactivity-of-a%C3%AE-plaque-associated-microglia-in-alzheimer-s-disease
#14
Zhuoran Yin, Divya Raj, Nasrin Saiepour, Debby Van Dam, Nieske Brouwer, Inge R Holtman, Bart J L Eggen, Thomas Möller, Joseph A Tamm, Aicha Abdourahman, Elly M Hol, Willem Kamphuis, Thomas A Bayer, Peter P De Deyn, Erik Boddeke
Alzheimer's disease (AD) is strongly associated with microglia-induced neuroinflammation. Particularly, Aβ plaque-associated microglia take on an "activated" morphology. However, the function and phenotype of these Aβ plaque-associated microglia are not well understood. We show hyperreactivity of Aβ plaque-associated microglia upon systemic inflammation in transgenic AD mouse models (i.e., 5XFAD and APP23). Gene expression profiling of Aβ plaque-associated microglia (major histocompatibility complex II(+) microglia) isolated from 5XFAD mice revealed a proinflammatory phenotype...
March 27, 2017: Neurobiology of Aging
https://www.readbyqxmd.com/read/28427424/microglial-depletion-alters-the-brain-neuroimmune-response-to-acute-binge-ethanol-withdrawal
#15
T Jordan Walter, Fulton T Crews
BACKGROUND: Recent studies have implicated microglia-the resident immune cells of the brain-in the pathophysiology of alcoholism. Indeed, post-mortem alcoholic brains show increased microglial markers and increased immune gene expression; however, the effects of ethanol on microglial functioning and how this impacts the brain remain unclear. In this present study, we investigate the effects of acute binge ethanol on microglia and how microglial depletion changes the brain neuroimmune response to acute binge ethanol withdrawal...
April 20, 2017: Journal of Neuroinflammation
https://www.readbyqxmd.com/read/28410218/6-shogaol-attenuates-lps-induced-inflammation-in-bv2-microglia-cells-by-activating-ppar-%C3%AE
#16
Qinghe Han, Qinghai Yuan, Xiaolin Meng, Junyuan Huo, Yuxin Bao, Guanghong Xie
6-Shogaol, a pungent agent isolated from Zingiber officinale Roscoe, has been known to have anti-tumor and anti-inflammatory effects. However, the anti-inflammatory effects and biological mechanism of 6-Shogaol in LPS-activated BV2 microglia remains largely unknown. In this study, we evaluated the anti-inflammatory effects of 6-Shogaol in LPS-activated BV2 microglia. 6-Shogaol was administrated 1 h before LPS treatment. The production of inflammatory mediators were detected by ELISA. The expression of NF-κB and PPAR-γ were detected by western blot analysis...
March 30, 2017: Oncotarget
https://www.readbyqxmd.com/read/28409447/isolation-of-peroxisomes-from-mouse-brain-using-a-continuous-nycodenz-gradient-a-comparison-to-the-isolation-of-liver-and-kidney-peroxisomes
#17
Miriam J Schönenberger, Werner J Kovacs
In the central nervous system (CNS) peroxisomes are present in all cell types, namely neurons, oligodendrocytes, astrocytes, microglia, and endothelial cells. Brain peroxisomes are smaller in size compared to peroxisomes from other tissues and are therefore referred to as microperoxisomes. We have established a purification procedure to isolate highly purified peroxisomes from the central nervous system that are well separated from the endoplasmic reticulum and mitochondria and are free of myelin contamination...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28402228/complement-components-are-expressed-by-infiltrating-macrophages-activated-microglia-early-following-viral-infection
#18
Jane E Libbey, Matthew F Cusick, Daniel J Doty, Robert S Fujinami
The individual innate immune components, interleukin-6 and complement component C3, play a role in the development of acute seizures in the Theiler's murine encephalomyelitis virus-induced seizure model. We examined the mRNA expression of various other complement components, cytokines, chemokines, and major histocompatibility complex antigens both within brain and in isolated ramified microglial and infiltrating macrophage/activated microglial cell populations over a time course covering the first 3 days postinfection...
April 12, 2017: Viral Immunology
https://www.readbyqxmd.com/read/28398520/injury-leads-to-the-appearance-of-cells-with-characteristics-of-both-microglia-and-astrocytes-in-mouse-and-human-brain
#19
Ulrika Wilhelmsson, Daniel Andersson, Yolanda de Pablo, Roy Pekny, Anders Ståhlberg, Jan Mulder, Nicholas Mitsios, Tibor Hortobágyi, Milos Pekny, Marcela Pekna
Microglia and astrocytes have been considered until now as cells with very distinct identities. Here, we assessed the heterogeneity within microglia/monocyte cell population in mouse hippocampus and determined their response to injury, by using single-cell gene expression profiling of cells isolated from uninjured and deafferented hippocampus. We found that in individual cells, microglial markers Cx3cr1, Aif1, Itgam, and Cd68 were co-expressed. Interestingly, injury led to the co-expression of the astrocyte marker Gfap in a subpopulation of Cx3cr1-expressing cells from both the injured and contralesional hippocampus...
April 7, 2017: Cerebral Cortex
https://www.readbyqxmd.com/read/28398146/protective-action-of-b1r-antagonist-against-cerebral-ischemia-reperfusion-injury-through-suppressing-mir-200c-expression-of-microglia-derived-microvesicles
#20
Min Tang, Ping Liu, Xia Li, Jian-Wen Wang, Xiong-Chao Zhu, Fang-Ping He
BACKGROUND AND OBJECTIVE: Cerebral ischemia-reperfusion (I/R) injury is a common side-effect for cerebral ischemic disease and its therapeutic regimen is limited. Kinin is pro-inflammatory peptide that is released and acts at the site of injury and inflammation such as brain and it works through bradykinin 1 receptor (B1R). The present study was to examine the effect of B1R antagonist on cerebral I/R injury and the potential mechanism. METHODS: Cerebral I/R injury was induced in mice by transient middle cerebral artery occlusion (MCAO)...
April 11, 2017: Neurological Research
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