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Astrocyte adult culture

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https://www.readbyqxmd.com/read/28154169/cd44-and-hyaluronan-regulate-adult-hippocampal-neural-stem-cell-quiescence-and-differentiation
#1
Weiping Su, Scott C Foster, Rubing Xing, Kerstin Feistel, Reid H J Olsen, Summer F Acevedo, Jacob Raber, Larry S Sherman
Adult neurogenesis in the hippocampal subgranular zone (SGZ) is involved in learning and memory throughout life but declines with aging. Mice lacking the CD44 transmembrane receptor for the glycosaminoglycan hyaluronan (HA) demonstrate a number of neurological disturbances including hippocampal memory deficits, implicating CD44 in the processes underlying hippocampal memory encoding, storage or retrieval. Here, we find that HA and CD44 play important roles in regulating adult neurogenesis, and we provide evidence that HA contributes to age-related reductions in neural stem cell (NSC) expansion and differentiation in the hippocampus...
January 31, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28151548/lack-of-appropriate-stoichiometry-strong-evidence-against-an-energetically-important-astrocyte-neuron-lactate-shuttle-in-brain
#2
REVIEW
Gerald A Dienel
Glutamate-stimulated aerobic glycolysis in astrocytes coupled with lactate shuttling to neurons where it can be oxidized was proposed as a mechanism to couple excitatory neuronal activity with glucose utilization (CMRglc ) during brain activation. From the outset, this model was not viable because it did not fulfill critical stoichiometric requirements: (i) Calculated glycolytic rates and measured lactate release rates were discordant in cultured astrocytes. (ii) Lactate oxidation requires oxygen consumption, but the oxygen-glucose index (OGI, calculated as CMRO2 /CMRglc ) fell during activation in human brain, and the small rise in CMRO2 could not fully support oxidation of lactate produced by disproportionate increases in CMRglc ...
February 2, 2017: Journal of Neuroscience Research
https://www.readbyqxmd.com/read/28099855/primary-cell-culture-of-live-neurosurgically-resected-aged-adult-human-brain-cells-and-single-cell-transcriptomics
#3
Jennifer M Spaethling, Young-Ji Na, Jaehee Lee, Alexandra V Ulyanova, Gordon H Baltuch, Thomas J Bell, Steven Brem, H Isaac Chen, Hannah Dueck, Stephen A Fisher, Marcela P Garcia, Mugdha Khaladkar, David K Kung, Timothy H Lucas, Donald M O'Rourke, Derek Stefanik, Jinhui Wang, John A Wolf, Tamas Bartfai, M Sean Grady, Jai-Yoon Sul, Junhyong Kim, James H Eberwine
Investigation of human CNS disease and drug effects has been hampered by the lack of a system that enables single-cell analysis of live adult patient brain cells. We developed a culturing system, based on a papain-aided procedure, for resected adult human brain tissue removed during neurosurgery. We performed single-cell transcriptomics on over 300 cells, permitting identification of oligodendrocytes, microglia, neurons, endothelial cells, and astrocytes after 3 weeks in culture. Using deep sequencing, we detected over 12,000 expressed genes, including hundreds of cell-type-enriched mRNAs, lncRNAs and pri-miRNAs...
January 17, 2017: Cell Reports
https://www.readbyqxmd.com/read/28099854/single-cell-transcriptomic-analysis-defines-heterogeneity-and-transcriptional-dynamics-in-the-adult-neural-stem-cell-lineage
#4
Ben W Dulken, Dena S Leeman, Stéphane C Boutet, Katja Hebestreit, Anne Brunet
Neural stem cells (NSCs) in the adult mammalian brain serve as a reservoir for the generation of new neurons, oligodendrocytes, and astrocytes. Here, we use single-cell RNA sequencing to characterize adult NSC populations and examine the molecular identities and heterogeneity of in vivo NSC populations. We find that cells in the NSC lineage exist on a continuum through the processes of activation and differentiation. Interestingly, rare intermediate states with distinct molecular profiles can be identified and experimentally validated, and our analysis identifies putative surface markers and key intracellular regulators for these subpopulations of NSCs...
January 17, 2017: Cell Reports
https://www.readbyqxmd.com/read/28060319/isolation-and-culture-of-adult-neural-stem-cells-from-the-mouse-subcallosal-zone
#5
Joo Yeon Kim, Ju-Hyun Lee, Woong Sun
Adult neural stem cells (aNSCs) can be used for the regeneration of damaged brain tissue. NSCs have the potential for differentiation and proliferation into three types of cells: neurons, astrocytes, and oligodendrocytes. Identifying aNSC-derived regions and characterizing the aNSC properties are critical for the potential use of aNSCs and for the elucidation of their role in neural regeneration. The subcallosal zone (SCZ), located between white matter and the hippocampus, has recently been reported to contain aNSCs and continuously give rise to neuroblasts...
December 15, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27987378/in-vitro-differentiation-of-neural-stem-cells-derived-from-human-olfactory-bulb-into-dopaminergic-like-neurons
#6
Rafieh Alizadeh, Gholamreza Hassanzadeh, Mohammad Taghi Joghataei, Mansoureh Soleimani, Fatemeh Moradi, Shahram Mohammadpour, Jahangir Ghorbani, Ali Safavi, Maryam Sarbishegi, Vahid Pirhajati Mahabadi, Leila Alizadeh, Mahmoudreza Hadjighassem
Obtaining new accessible source of neuronal stem cells that can be used in Parkinson's disease cell transplant is the aim of this research. The human olfactory bulb contains neural stem cells (NSCs) that are responsible for the neurogenesis in the brain and the replacement of damaged cellular components throughout life. NSCs are capable of differentiating into neuronal and glial cells. We isolated NSCs from brain-death donor's olfactory bub and differentiate them to dopaminergic neurons. The obtained olfactory bulb tissues were cultured in DMEM12, B27 supplemented with bFGF, EGF, and LIF...
December 17, 2016: European Journal of Neuroscience
https://www.readbyqxmd.com/read/27932954/neuronal-release-of-cytokine-il-3-triggered-by-mechanosensitive-autostimulation-of-the-p2x7-receptor-is-neuroprotective
#7
Jason C Lim, Wennan Lu, Jonathan M Beckel, Claire H Mitchell
Mechanical strain due to increased pressure or swelling activates inflammatory responses in many neural systems. As cytokines and chemokine messengers lead to both pro-inflammatory and neuroprotective actions, understanding the signaling patterns triggered by mechanical stress may help improve overall outcomes. While cytokine signaling in neural systems is often associated with glial cells like astrocytes and microglia, the contribution of neurons themselves to the cytokine response is underappreciated and has bearing on any balanced response...
2016: Frontiers in Cellular Neuroscience
https://www.readbyqxmd.com/read/27911416/the-indirect-neuron-astrocyte-coculture-assay-an-in-vitro-set-up-for-the-detailed-investigation-of-neuron-glia-interactions
#8
Christine Gottschling, Egor Dzyubenko, Maren Geissler, Andreas Faissner
Proper neuronal development and function is the prerequisite of the developing and the adult brain. However, the mechanisms underlying the highly controlled formation and maintenance of complex neuronal networks are not completely understood thus far. The open questions concerning neurons in health and disease are diverse and reaching from understanding the basic development to investigating human related pathologies, e.g., Alzheimer's disease and Schizophrenia. The most detailed analysis of neurons can be performed in vitro...
November 14, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27873129/expression-of-dixdc1-and-its-role-in-astrocyte-proliferation-after-traumatic-brain-injury
#9
Hongjian Lu, Rui Jiang, Xuelei Tao, Chengwei Duan, Jie Huang, Wei Huan, Yunfen He, Jianbin Ge, Jianbing Ren
DIX domain containing 1 (Dixdc1), a positive regulator of Wnt signaling pathway, is recently reported to play a role in the neurogenesis. However, the distribution and function of Dixdc1 in the central nervous system (CNS) after brain injury are still unclear. We used an acute traumatic brain injury (TBI) model in adult rats to investigate whether Dixdc1 is involved in CNS injury and repair. Western blot analysis and immunohistochemistry showed a time-dependent up-regulation of Dixdc1 expression in ipsilateral cortex after TBI...
November 21, 2016: Cellular and Molecular Neurobiology
https://www.readbyqxmd.com/read/27822179/plasmid-based-generation-of-induced-neural-stem-cells-from-adult-human-fibroblasts
#10
Philipp Capetian, Luis Azmitia, Martje G Pauly, Victor Krajka, Felix Stengel, Eva-Maria Bernhardi, Mariana Klett, Britta Meier, Philip Seibler, Nancy Stanslowsky, Andreas Moser, Andreas Knopp, Gabriele Gillessen-Kaesbach, Guido Nikkhah, Florian Wegner, Máté Döbrössy, Christine Klein
Direct reprogramming from somatic to neural cell types has become an alternative to induced pluripotent stem cells. Most protocols employ viral expression systems, posing the risk of random genomic integration. Recent developments led to plasmid-based protocols, lowering this risk. However, these protocols either relied on continuous presence of a variety of small molecules or were only able to reprogram murine cells. We therefore established a reprogramming protocol based on vectors containing the Epstein-Barr virus (EBV)-derived oriP/EBNA1 as well as the defined expression factors Oct3/4, Sox2, Klf4, L-myc, Lin28, and a small hairpin directed against p53...
2016: Frontiers in Cellular Neuroscience
https://www.readbyqxmd.com/read/27815898/synchronization-of-mammalian-cell-cultures-by-serum-deprivation
#11
Thomas J Langan, Kyla R Rodgers, Richard C Chou
Mammalian cells are amenable to the study of regulatory mechanisms dictating cell cycle progression in vitro by shifting them into the same phase of the cycle. Procedures to arrest cultured cells in specific phases of the cell cycle may be termed in vitro synchronization. The procedure described here was developed for the study of primary astrocytes and a glioma cell line, but is broadly applicable to other mammalian cells. Its application allows astrocytes to re-enter the cell cycle from a state of quiescence (G0) under carefully defined experimental conditions to move together into subsequent phases such as the G1 and S phases...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27798130/mechanisms-of-co2-h-sensitivity-of-astrocytes
#12
Egor Turovsky, Shefeeq M Theparambil, Vitaliy Kasymov, Joachim W Deitmer, Ana Gutierrez Del Arroyo, Gareth L Ackland, Jason J Corneveaux, April N Allen, Matthew J Huentelman, Sergey Kasparov, Nephtali Marina, Alexander V Gourine
: Ventral regions of the medulla oblongata of the brainstem are populated by astrocytes sensitive to physiological changes in PCO2/[H(+)]. These astrocytes respond to decreases in pH with elevations in intracellular Ca(2+) and facilitated exocytosis of ATP-containing vesicles. Released ATP propagates Ca(2+) excitation among neighboring astrocytes and activates neurons of the brainstem respiratory network triggering adaptive increases in breathing. The mechanisms linking increases in extracellular and/or intracellular PCO2/[H(+)] with Ca(2+) responses in chemosensitive astrocytes remain unknown...
October 19, 2016: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/27758000/cell-autonomous-and-noncell-autonomous-role-of-nf-%C3%AE%C2%BAb-p50-in-astrocyte-mediated-fate-specification-of-adult-neural-progenitor-cells
#13
Suzana Cvijetic, Valeria Bortolotto, Marcello Manfredi, Elia Ranzato, Emilio Marengo, Rita Salem, Pier Luigi Canonico, Mariagrazia Grilli
In previous work, we demonstrated that NF-κB p50 acts as crucial regulator of adult hippocampal neural progenitor cells (ahNPC). Indeed, NF-κB p50 knockout (KO) mice are characterized by remarkably reduced hippocampal neurogenesis. As a follow up to that work, herein we show that when cultured in vitro, ahNPC from wild type (WT) and p50KO mice are not significantly different in their neurogenic potential. This observation prompted us to investigate cell-autonomous and noncell-autonomous consequences of p50 absence on neuronal fate specification of ahNPC...
October 19, 2016: Glia
https://www.readbyqxmd.com/read/27726178/critical-role-of-ror2-receptor-tyrosine-kinase-in-regulating-cell-cycle-progression-of-reactive-astrocytes-following-brain-injury
#14
Mitsuharu Endo, Guljahan Ubulkasim, Chiho Kobayashi, Reiko Onishi, Atsu Aiba, Yasuhiro Minami
Ror2 receptor tyrosine kinase plays crucial roles in developmental morphogenesis and tissue-/organo-genesis. In the developing brain, Ror2 is expressed in neural stem/progenitor cells (NPCs) and involved in the regulation of their stemness. However, it remains largely unknown about its role in the adult brain. In this study, we show that Ror2 is up-regulated in reactive astrocytes in the neocortices within 3 days following stab-wound injury. Intriguingly, Ror2-expressing astrocytes were detected primarily at the area surrounding the injury site, where astrocytes express Nestin, a marker of NPCs, and proliferate in response to injury...
October 11, 2016: Glia
https://www.readbyqxmd.com/read/27666792/hedgehog-controls-quiescence-and-activation-of-neural-stem-cells-in-the-adult-ventricular-subventricular-zone
#15
Mathieu Daynac, Linda Tirou, Hélène Faure, Marc-André Mouthon, Laurent R Gauthier, Heidi Hahn, François D Boussin, Martial Ruat
Identifying the mechanisms controlling quiescence and activation of neural stem cells (NSCs) is crucial for understanding brain repair. Here, we demonstrate that Hedgehog (Hh) signaling actively regulates different pools of quiescent and proliferative NSCs in the adult ventricular-subventricular zone (V-SVZ), one of the main brain neurogenic niches. Specific deletion of the Hh receptor Patched in NSCs during adulthood upregulated Hh signaling in quiescent NSCs, progressively leading to a large accumulation of these cells in the V-SVZ...
October 11, 2016: Stem Cell Reports
https://www.readbyqxmd.com/read/27602084/comparative-effect-of-immature-neuronal-or-glial-cell-transplantation-on-motor-functional-recovery-following-experimental-traumatic-brain-injury-in-rats
#16
Fu-Shi Quan, Jian Chen, Yuan Zhong, Wen-Zhi Ren
The present study evaluated the comparative effect of stereotaxically transplanted immature neuronal or glial cells in brain on motor functional recovery and cytokine expression after cold-induced traumatic brain injury (TBI) in adult rats. A total of 60 rats were divided into four groups (n=15/group): Sham group; TBI only group; TBI plus neuronal cells-transplanted group (NC-G); and TBI plus glial cells-transplanted group (GC-G). Cortical lesions were induced by a touching metal stamp, frozen with liquid nitrogen, to the dura mater over the motor cortex of adult rats...
September 2016: Experimental and Therapeutic Medicine
https://www.readbyqxmd.com/read/27585449/anti-aging-effects-of-guanosine-in-glial-cells
#17
Débora Guerini Souza, Bruna Bellaver, Larissa Daniele Bobermin, Diogo Onofre Souza, André Quincozes-Santos
Guanosine, a guanine-based purine, has been shown to exert beneficial roles in in vitro and in vivo injury models of neural cells. Guanosine is released from astrocytes and modulates important astroglial functions, including glutamatergic metabolism, antioxidant, and anti-inflammatory activities. Astrocytes are crucial for regulating the neurotransmitter system and synaptic information processes, ionic homeostasis, energy metabolism, antioxidant defenses, and the inflammatory response. Aging is a natural process that induces numerous changes in the astrocyte functionality...
December 2016: Purinergic Signalling
https://www.readbyqxmd.com/read/27564458/a-comparative-transcriptomic-analysis-of-astrocytes-differentiation-from-human-neural-progenitor-cells
#18
Marco Magistri, Nathalie Khoury, Emilia Maria Cristina Mazza, Dmitry Velmeshev, Jae K Lee, Silvio Bicciato, Pantelis Tsoulfas, Mohammad Ali Faghihi
Astrocytes are a morphologically and functionally heterogeneous population of cells that play critical roles in neurodevelopment and in the regulation of central nervous system homeostasis. Studies of human astrocytes have been hampered by the lack of specific molecular markers and by the difficulties associated with purifying and culturing astrocytes from adult human brains. Human neural progenitor cells (NPCs) with self-renewal and multipotent properties represent an appealing model system to gain insight into the developmental genetics and function of human astrocytes, but a comprehensive molecular characterization that confirms the validity of this cellular system is still missing...
November 2016: European Journal of Neuroscience
https://www.readbyqxmd.com/read/27544484/astrocytes-from-old-alzheimer-s-disease-mice-are-impaired-in-a%C3%AE-uptake-and-in-neuroprotection
#19
Tal Iram, Dorit Trudler, David Kain, Sivan Kanner, Ronit Galron, Robert Vassar, Ari Barzilai, Pablo Blinder, Zvi Fishelson, Dan Frenkel
In Alzheimer's disease (AD), astrocytes undergo morphological changes ranging from atrophy to hypertrophy, but the effect of such changes at the functional level is still largely unknown. Here, we aimed to investigate whether alterations in astrocyte activity in AD are transient and depend on their microenvironment, or whether they are irreversible. We established and characterized a new protocol for the isolation of adult astrocytes and discovered that astrocytes isolated from old 5xFAD mice have higher GFAP expression than astrocytes derived from WT mice, as observed in vivo...
August 17, 2016: Neurobiology of Disease
https://www.readbyqxmd.com/read/27470924/quantification-of-the-functional-expression-of-the-ca-2-activated-k-channel-kca-3-1-on-microglia-from-adult-human-neocortical-tissue
#20
Linda V Blomster, Dorte Strøbaek, Charlotte Hougaard, Jessica Klein, Lars H Pinborg, Jens D Mikkelsen, Palle Christophersen
The KCa 3.1 channel (KCNN4) is an important modulator of microglia responses in rodents, but no information exists on functional expression on microglia from human adults. We isolated and cultured microglia (max 1% astrocytes, no neurons or oligodendrocytes) from neocortex surgically removed from epilepsy patients and employed electrophysiological whole-cell measurements and selective pharmacological tools to elucidate functional expression of KCa 3.1. The channel expression was demonstrated as a significant increase in the voltage-independent current by NS309, a KCa 3...
July 29, 2016: Glia
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