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Isolation microglia

Nynke Oosterhof, Inge R Holtman, Laura E Kuil, Herma C van der Linde, Erik W G M Boddeke, Bart J L Eggen, Tjakko J van Ham
Microglia are brain resident macrophages important for brain development, connectivity, homeostasis and disease. However, it is still largely unclear how microglia functions and their identity are regulated at the molecular level. Although recent transcriptomic studies have identified genes specifically expressed in microglia, the function of most of these genes in microglia is still unknown. Here, we performed RNA sequencing on microglia acutely isolated from healthy and neurodegenerative zebrafish brains...
October 19, 2016: Glia
Daiki Yamanaka, Takashi Kawano, Atsushi Nishigaki, Bun Aoyama, Hiroki Tateiwa, Marie Shigematsu-Locatelli, Fabricio M Locatelli, Masataka Yokoyama
PURPOSE: In the present study, we examined whether and by what mechanisms dexmedetomidine (DMED) prevents the development of systemic inflammation (SI)-induced cognitive dysfunction in aged rats. METHODS: Animals received a single intraperitoneal (i.p.) injection of either 5.0 mg/kg lipopolysaccharide (LPS) or vehicle. LPS-treated rats were further divided into three groups: early DMED, late DMED, or midazolam (MDZ) treatment (n = 12 each). Seven days after LPS injection, cognitive function was evaluated using a novel object recognition task, followed by measurement of hippocampal levels of proinflammatory cytokines and Toll-like receptor 4 (TLR-4) expression...
October 13, 2016: Journal of Anesthesia
Marija Adzic, Ivana Stevanovic, Natasa Josipovic, Danijela Laketa, Irena Lavrnja, Ivana M Bjelobaba, Iva Bozic, Marija Jovanovic, Milena Milosevic, Nadezda Nedeljkovic
It is widely accepted that adenosine triphosphate (ATP) acts as a universal danger-associated molecular pattern with several known mechanisms for immune cell activation. In the central nervous system, ATP activates microglia and astrocytes and induces a neuroinflammatory response. The aim of the present study was to describe responses of isolated astrocytes to increasing concentrations of ATP (5 µM to 1 mM), which were intended to mimic graded intensity of the extracellular stimulus. The results show that ATP induces graded activation response of astrocytes in terms of the cell proliferation, stellation, shape remodeling, and underlying actin and GFAP filament rearrangement, although the changes occurred without an apparent increase in GFAP and actin protein expression...
October 7, 2016: Journal of Neuroscience Research
Bernardo Castellano, Mar Bosch-Queralt, Beatriz Almolda, Nàdia Villacampa, Berta González
Microglial cells are highly dynamic cells with processes continuously moving to survey the surrounding territory. Microglia possess a broad variety of surface receptors and subtle changes in their microenvironment cause microglial cell processes to extend, retract, and interact with neuronal synaptic contacts. When the nervous system is disturbed, microglia activate, proliferate, and migrate to sites of injury in response to alert signals. Released nucleotides like ATP and UTP are among the wide range of molecules promoting microglial activation and guiding their migration and phagocytic function...
2016: Advances in Experimental Medicine and Biology
Chung Sub Kim, Lalita Subedi, Oh Wook Kwon, Hyun Bong Park, Sun Yeou Kim, Sang Un Choi, Kang Ro Lee
Five new lignan glycosides, wasabisides A-E (1-5), and four known phenolic compounds (6-9), were isolated from the roots of Wasabia japonica. The chemical structures of the new compounds (1-5) were determined through spectroscopic analysis and chemical methods. All isolated compounds (1-9) were evaluated for their potential neuroprotective effects through induction of nerve growth factor in C6 glioma cells, for their effects on nitric oxide levels in lipopolysaccharide-stimulated murine microglia BV2 cells, and for their cytotoxicity against four human tumor cell lines (A549, SK-OV-3, SK-MEL-2, and BT549)...
October 4, 2016: Journal of Natural Products
Arjun Saha, Susan Buntz, Paula Scotland, Li Xu, Pamela Noeldner, Sachit Patel, Amy Wollish, Aruni Gunaratne, Tracy Gentry, Jesse Troy, Glenn K Matsushima, Joanne Kurtzberg, Andrew E Balber
Microglia and monocytes play important roles in regulating brain remyelination. We developed DUOC-01, a cell therapy product intended for treatment of demyelinating diseases, from banked human umbilical cord blood (CB) mononuclear cells. Immunodepletion and selection studies demonstrated that DUOC-01 cells are derived from CB CD14(+) monocytes. We compared the ability of freshly isolated CB CD14(+) monocytes and DUOC-01 cells to accelerate remyelination of the brains of NOD/SCID/IL2Rγ(null) mice following cuprizone feeding-mediated demyelination...
August 18, 2016: JCI Insight
Laura Korvers, Amanda de Andrade Costa, Martin Mersch, Vitali Matyash, Helmut Kettenmann, Marcus Semtner
Microglia are the resident immune cells in the central nervous system and many of their physiological functions are known to be linked to intracellular calcium (Ca(2+)) signaling. Here we show that isolated and purified mouse microglia-either freshly or cultured-display spontaneous and transient Ca(2+) elevations lasting for around ten to twenty seconds and occurring at frequencies of around five to ten events per hour and cell. The events were absent after depletion of internal Ca(2+) stores, by phospholipase C (PLC) inhibition or blockade of inositol-1,4,5-trisphosphate receptors (IP3Rs), but not by removal of extracellular Ca(2+), indicating that Ca(2+) is released from endoplasmic reticulum intracellular stores...
September 22, 2016: Cell Calcium
Shahnawaz Ali Bhat, Ruby Goel, Rakesh Shukla, Kashif Hanif
Studies have demonstrated separately that hypertension is associated with platelet activation in the periphery (resulting in accumulation and localized inflammatory response) and glial activation in the brain. We investigated the contribution of platelets in brain inflammation, particularly glial activation in vitro and in a rat model of hypertension. We found that HTN increased the expression of adhesion molecules like JAM-1, ICAM-1, and VCAM-1 on brain endothelium and resulted in the deposition of platelets in the brain...
September 19, 2016: Brain, Behavior, and Immunity
Adrienne M Antonson, Emily C Radlowski, Marcus A Lawson, Jennifer L Rytych, Rodney W Johnson
Maternal infection during pregnancy increases risk for neurodevelopmental disorders and reduced stress resilience in offspring, but the mechanisms are not fully understood. We hypothesized that piglets born from gilts infected with a respiratory virus during late gestation would exhibit aberrant microglia activity, cognitive deficits and reduced sociability. Pregnant gilts were inoculated with porcine reproductive and respiratory syndrome virus (PRRSV; 5 x 10(5) TCID50 of live PRRSV) or saline at gestational day 76...
September 17, 2016: Brain, Behavior, and Immunity
Chi-Su Yoon, Dong-Cheol Kim, Tran Hong Quang, Jungwon Seo, Dae Gill Kang, Ho Sub Lee, Hyuncheol Oh, Youn-Chul Kim
Cudrania tricuspidata Bureau (Moraceae) is an important source of traditional Korean and Chinese medicines used to treat neuritis and inflammation. Cudratricusxanthone A (1), a prenylated xanthone, isolated from C. tricuspidata, has a variety of biological and therapeutic activities. The goal of this study was to examine the effects of compound 1 on neuroinflammation and characterize its mechanism of action in lipopolysaccharide (LPS)-stimulated BV2 microglia. Cudratricusxanthone A (1) suppressed the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 enzymes and decreased the production of iNOS-derived nitric oxide and COX-2-derived prostaglandin E2 in LPS-stimulated mouse BV2 microglia...
2016: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
Jae Sik Yu, Jiwon Baek, Hyun Bong Park, Eunjung Moon, Sun Yeou Kim, Sang Un Choi, Ki Hyun Kim
A new rearranged eudesmane sesquiterpene, named eudeglaucone (1), and five known sesquiterpenes including (+)-faurinone (2) and four eudesmane-type sesquiterpenes (3-6), were isolated from the twigs of Lindera glauca (Sieb. et Zucc.) Blume. The structure of 1 was elucidated by a combination of extensive spectroscopic analyses, including extensive 2D NMR ((1)H-(1)H COSY, HMQC, HMBC, and NOESY) and HR-MS. Compound 1 was a relatively rare rearranged eudesmane sesquiterpene in terpenoids. All isolates were evaluated for their antiproliferative activities against four human tumor cell lines (A549, SK-OV-3, SK-MEL-2, and HCT-15)...
September 12, 2016: Archives of Pharmacal Research
Nicholas H Varvel, Jonas J Neher, Andrea Bosch, Wenyi Wang, Richard M Ransohoff, Richard J Miller, Raymond Dingledine
The generalized seizures of status epilepticus (SE) trigger a series of molecular and cellular events that produce cognitive deficits and can culminate in the development of epilepsy. Known early events include opening of the blood-brain barrier (BBB) and astrocytosis accompanied by activation of brain microglia. Whereas circulating monocytes do not infiltrate the healthy CNS, monocytes can enter the brain in response to injury and contribute to the immune response. We examined the cellular components of innate immune inflammation in the days following SE by discriminating microglia vs...
September 20, 2016: Proceedings of the National Academy of Sciences of the United States of America
Dong-Sung Lee, Mina Lee, Sang Hyun Sung, Gil Saeng Jeong
Glutamate-induced oxidative injury contributes to neuronal degeneration such as Alzheimer's disease, Parkinson's disease and Huntington's disease in the central nervous system (CNS). Siegesbeckia pubescens is used in oriental medicine to treat arthritis, stroke, rash, edema, and skin itching eczema in South-East Asia. This study provides evidence that 5,3'-dihydroxy-3,7,4'-trimethoxyflavone (DTMF), a compound isolated from 90% MeOH extract of Siegesbeckia pubescens, effectively has neuroprotective and anti-neuroinflammatory activities...
August 29, 2016: International Immunopharmacology
Laura K Fonken, Meagan M Kitt, Andrew D Gaudet, Ruth M Barrientos, Linda R Watkins, Steven F Maier
Aged animals exhibit diminished circadian rhythms, and both aging and circadian disruption sensitize neuroinflammatory responses. Microglia-the innate immune cell of the central nervous system-possess endogenous timekeeping mechanisms that regulate immune responses. Here, we explored whether aging is associated with disrupted diurnal rhythms in microglia and neuroinflammatory processes. First, hippocampal microglia isolated from young rats (4 months F344XBN) rhythmically expressed circadian clock genes, whereas microglia isolated from the hippocampus of aged rats (25 months) had aberrant Per1 and Per2 rhythms...
August 1, 2016: Neurobiology of Aging
Ling Guo, Aras Rezvanian, Lokesh Kukreja, Ramez Hoveydai, Eileen H Bigio, M-Marsel Mesulam, Joseph El Khoury, Changiz Geula
Microglia are immune cells of the brain that display a range of functions. Most of our knowledge about microglia biology and function is based on cells from the rodent brain. Species variation in the complexity of the brain and differences in microglia response in the primate when compared with the rodent, require use of adult human microglia in studies of microglia biology. While methods exist for isolation of microglia from postmortem human brains, none allow culturing cells to high passage. Thus cells from the same case could not be used in parallel studies and multiple conditions...
October 4, 2016: Journal of Alzheimer's Disease: JAD
Zhiqian Yu, Hotaka Fukushima, Chiaki Ono, Mai Sakai, Yoshiyuki Kasahara, Yoshie Kikuchi, Nicole Gunawansa, Yuta Takahashi, Hiroo Matsuoka, Satoshi Kida, Hiroaki Tomita
The proinflammatory cytokine productions in the brain are altered in a process of fear memory formation, indicating a possibility that altered microglial function may contribute to fear memory formation. We aimed to investigate whether and how microglial function contributes to fear memory formation. Expression levels of M1- and M2-type microglial marker molecules in microglia isolated from each conditioned mice group were assessed by real-time PCR and immunohistochemistry. Levels of tumor necrosis factor (TNF)-α, but not of other proinflammatory cytokines produced by M1-type microglia, increased in microglia from mice representing retention of fear memory, and returned to basal levels in microglia from mice representing extinction of fear memory...
August 22, 2016: Brain, Behavior, and Immunity
Takashi Kawano, Hideki Iwata, Bun Aoyama, Atsushi Nishigaki, Daiki Yamanaka, Hiroki Tateiwa, Satoru Eguchi, Fabricio M Locatelli, Masataka Yokoyama
AIMS: This study aimed to investigate the role of central insulin signaling, including glycogen synthase kinase 3β (GSK-3β), and its therapeutic potential for the prevention of postoperative neurocognitive deficits. MAIN METHODS: In non-insulin experiment, aged rats were divided into a sham group and abdominal surgery group. In insulin experiment, sham and surgically treated rats were distributed into two groups: an intranasal denatured insulin-treated group and intranasal insulin-treated group...
October 1, 2016: Life Sciences
Matthew McMillin, Stephanie Grant, Gabriel Frampton, Sarah Andry, Adam Brown, Sharon DeMorrow
BACKGROUND: Acute liver failure is associated with numerous systemic consequences including neurological dysfunction, termed hepatic encephalopathy, which contributes to mortality and is a challenge to manage in the clinic. During hepatic encephalopathy, microglia activation and neuroinflammation occur due to dysregulated cell signaling and an increase of toxic metabolites in the brain. Fractalkine is a chemokine that is expressed primarily in neurons and through signaling with its receptor CX3CR1 on microglia, leads to microglia remaining in a quiescent state...
2016: Journal of Neuroinflammation
Liqing Zhang, Zhangang Zhang, Yan Fu, Pin Yang, Zhenxia Qin, Yongjun Chen, Ying Xu
Microglia activation and neuroinflammation are critically involved in pathogenesis of neurodegenerative disorders. Patients with neurodegenerative disorders often suffer memory impairment and currently there is no effective treatment for inflammation-led memory impairment. Trans-cinnamaldehyde (TCA) isolated from medicinal herb Cinnamomum cassia has been shown to exhibit anti-inflammatory capability. However, the potential of TCA to be used to improve memory impairment under neuroinflammation has not been explored...
November 2016: Neuropharmacology
Pratima Rawat, Stephen A Spector
Microglia cells are the major reservoir of HIV-1 (HIV) within the CNS. However, current models using transformed cell lines are not representative of primary microglia and fetal brain samples for isolation of primary human microglia (HMG) are increasingly difficult to obtain. Here, we describe a monocyte-derived microglia (MMG) cell model of HIV infection that recapitulates infection of primary HMG. CD14(+) cells isolated from healthy donors were cultured with M-CSF, beta-nerve growth factor, GM-CSF, and CCL2, and compared to HMG...
August 18, 2016: Journal of Neurovirology
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