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Astrocyte adult culture

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https://www.readbyqxmd.com/read/28381236/microglia-activation-is-essential-for-bmp7-mediated-retinal-reactive-gliosis
#1
Subramanian Dharmarajan, Debra L Fisk, Christine M Sorenson, Nader Sheibani, Teri L Belecky-Adams
BACKGROUND: Our previous studies have shown that BMP7 is able to trigger activation of retinal macroglia. However, these studies showed the responsiveness of Müller glial cells and retinal astrocytes in vitro was attenuated in comparison to those in vivo, indicating other retinal cell types may be mediating the response of the macroglial cells to BMP7. In this study, we test the hypothesis that BMP7-mediated gliosis is the result of inflammatory signaling from retinal microglia. METHODS: Adult mice were injected intravitreally with BMP7 and eyes harvested 1, 3, or 7 days postinjection...
April 5, 2017: Journal of Neuroinflammation
https://www.readbyqxmd.com/read/28374892/in-vitro-adult-astrocytes-are-derived-from-mature-cells-and-reproduce-in-vivo-redox-profile
#2
Débora Guerini Souza, Bruna Bellaver, Silvia Resende Terra, Fatima Costa Rodrigues Guma, Diogo Onofre Souza, André Quincozes-Santos
Astrocytes are versatile cells involved in synaptic information processing, energy metabolism, redox homeostasis, inflammatory response and structural support of the brain. Recently, we established a routine protocol of cultured astrocytes derived from adult and aged Wistar rats, which present several different responses compared to newborn astrocytes, commonly used to characterize the role of the astrocytes in the central nervous system. Previous studies hypothesized that astrocyte cultures prepared from adult animals derive from immature precursors present in the adult tissue throughout life...
April 4, 2017: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/28295574/cd38-positively-regulates-postnatal-development-of-astrocytes-cell-autonomously-and-oligodendrocytes-non-cell-autonomously
#3
Tsuyoshi Hattori, Minoru Kaji, Hiroshi Ishii, Roboon Jureepon, Mika Takarada-Iemata, Hieu Minh Ta, Thuong Manh Le, Ayumu Konno, Hirokazu Hirai, Yoshitake Shiraishi, Noriyuki Ozaki, Yasuhiko Yamamoto, Hiroshi Okamoto, Shigeru Yokoyama, Haruhiro Higashida, Yasuko Kitao, Osamu Hori
Glial development is critical for the function of the central nervous system. CD38 is a multifunctional molecule with ADP-ribosyl cyclase activity. While critical roles of CD38 in the adult brain such as oxytocin release and social behavior have been reported, those in the developing brain remain largely unknown. Here we demonstrate that deletion of Cd38 leads to impaired development of astrocytes and oligodendrocytes in mice. CD38 is highly expressed in the developing brains between postnatal day 14 (P14) and day 28 (P28)...
March 13, 2017: Glia
https://www.readbyqxmd.com/read/28255907/homocysteine-induces-glial-reactivity-in-adult-rat-astrocyte-cultures
#4
Aline Longoni, Bruna Bellaver, Larissa Daniele Bobermin, Camila Leite Santos, Yasmine Nonose, Janaina Kolling, Tiago M Dos Santos, Adriano M de Assis, André Quincozes-Santos, Angela T S Wyse
Astrocytes are dynamic glial cells associated to neurotransmitter systems, metabolic functions, antioxidant defense, and inflammatory response, maintaining the brain homeostasis. Elevated concentrations of homocysteine (Hcy) are involved in the pathogenesis of age-related neurodegenerative disorders, such as Parkinson and Alzheimer diseases. In line with this, our hypothesis was that Hcy could promote glial reactivity in a model of cortical primary astrocyte cultures from adult Wistar rats. Thus, cortical astrocytes were incubated with different concentrations of Hcy (10, 30, and 100 μM) during 24 h...
March 2, 2017: Molecular Neurobiology
https://www.readbyqxmd.com/read/28250752/neural-differentiation-of-choroid-plexus-epithelial-cells-role-of-human-traumatic-cerebrospinal-fluid
#5
Elham Hashemi, Yousef Sadeghi, Abbas Aliaghaei, Afsoun Seddighi, Abbas Piryaei, Mehdi Eskandarian Broujeni, Fatemeh Shaerzadeh, Abdollah Amini, Ramin Pouriran
As the key producer of cerebrospinal fluid (CSF), the choroid plexus (CP) provides a unique protective system in the central nervous system. CSF components are not invariable and they can change based on the pathological conditions of the central nervous system. The purpose of the present study was to assess the effects of non-traumatic and traumatic CSF on the differentiation of multipotent stem-like cells of CP into the neural and/or glial cells. CP epithelial cells were isolated from adult male rats and treated with human non-traumatic and traumatic CSF...
January 2017: Neural Regeneration Research
https://www.readbyqxmd.com/read/28154169/cd44-transmembrane-receptor-and-hyaluronan-regulate-adult-hippocampal-neural-stem-cell-quiescence-and-differentiation
#6
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 found 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...
March 17, 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
#7
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
#8
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
#9
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
#10
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
#11
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
This study describes a new accessible source of neuronal stem cells that can be used in Parkinson's disease cell transplant. The human olfactory bulb contains neural stem cells (NSCs) that are responsible for 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 the olfactory bulb of brain-death donors and differentiated them into dopaminergic neurons. The olfactory bulb tissues obtained were cultured in Dulbecco's modified Eagle's medium/nutrient mixture F12, B27 supplemented with basic fibroblast growth factor, epidermal growth factor and leukemia inhibitory factor...
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
#12
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
#13
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
#14
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
#15
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
#16
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
#17
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
#18
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
#19
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
#20
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
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