The Astrocyte Network in the Ventral Nerve Cord Neuropil of the Drosophila Third-Instar Larva
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In Vivo Imaging of Microglia-Mediated Axonal Pruning and Modulation By
Combined bioRxivsingle preprintmanuscript doi: https://doi.org/10.1101/2020.06.07.087221 file ; this version posted June 8, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 1 In vivo imaging of microglia-mediated axonal pruning and modulation 2 by the complement system 3 Tony K.Y. Lim1 and Edward S. Ruthazer1,2,* 4 1. Department of Neurology & Neurosurgery, Montreal Neurological Institute-Hospital, McGill 5 University, Montreal, Quebec, H3A 2B4; Canada 6 2. Lead Contact 7 *Correspondence: [email protected] 8 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.06.07.087221; this version posted June 8, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 9 Summary 10 Partial phagocytosis – called trogocytosis – of axons by microglia has been documented in ex vivo 11 preparations but has yet to be observed in vivo. Fundamental questions regarding the mechanisms that 12 modulate axon trogocytosis as well as its function in neural circuit development remain unanswered. 13 Here we used 2-photon live imaging of the developing Xenopus laevis retinotectal circuit to observe 14 axon trogocytosis by microglia in vivo. Amphibian regulator of complement activation 3 (aRCA3) was 15 identified as a neuronally expressed, synapse-associated complement inhibitory molecule. 16 Overexpression of aRCA3 enhanced axonal arborization and inhibited trogocytosis, while expression of 17 VAMP2-C3, a complement-enhancing fusion protein tethered to the axon surface, reduced axonal 18 arborization. -
Microglia Control Glutamatergic Synapses in the Adult Mouse Hippocampus
bioRxiv preprint doi: https://doi.org/10.1101/2021.02.01.429096; this version posted February 2, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Microglia control glutamatergic synapses in the adult mouse hippocampus Short title: Microglia and glutamatergic synapses Bernadette Basilico1†*‡, Laura Ferrucci1‡, Patrizia Ratano2‡, Maria T. Golia1, Alfonso Grimaldi3, Maria Rosito3, Valentina Ferretti4, Ingrid Reverte1,5, Maria C. Marrone6, Maria Giubettini3,7, Valeria De Turris3, Debora Salerno3, Stefano Garofalo1, Marie-Kim St-Pierre8, Micael Carrier8, Massimiliano Renzi1, Francesca Pagani3, Marcello Raspa9, Ferdinando Scavizzi9, Cornelius T. Gross10, Silvia Marinelli5, Marie E. Tremblay8,11, Daniele Caprioli1,5, Laura Maggi1, Cristina Limatola1,2, Silvia Di Angelantonio1,3§, Davide Ragozzino1,5*§ 1Department of Physiology and Pharmacology, Sapienza University of Rome, Rome, Italy. 2IRCCS Neuromed, Via Atinese 18, 86077, Pozzilli, IS, Italy. 3Center for Life Nanoscience, Istituto Italiano di Tecnologia, Rome, Italy. 4Dipartimento di Biologia e Biotecnologie "Charles Darwin", Sapienza University of Rome, Rome, Italy. 5Santa Lucia Foundation (IRCCS Fondazione Santa Lucia), Rome, Italy. 6European Brain Research Institute-Rita Levi Montalcini, Rome, Italy. 7CrestOptics S.p.A., Via di Torre Rossa 66, 00165 Rome, Italy. 8Centre de Recherche du CHU de Québec, Axe Neurosciences Québec, QC, Canada; Département de médecine moléculaire, Université Laval Québec, QC, Canada. 9National Research Council, Institute of Biochemistry and Cell Biology (CNR- IBBC/EMMA/Infrafrontier/IMPC), International Campus “A. Buzzati-Traverso”, Monterotondo (Rome) Italy. -
Astrocyte Cell Culture Preparation of Flasks: 1
Astrocyte Cell Culture Preparation of flasks: 1. Coat T75 flask(s) with 1 mg/ml of PureCol (Collagen) overnight 2. Remove solution, rinse flasks with sterile ddH20, set the flasks upright and allow to dry in culture hood for 2 hr Dissection: 1. Dissect P1-P3 pups: Remove brainstem, cerebellum and diencephalons in cold dissection buffer. Peel off meninges and transfer cortex to a 50 ml tube on ice, which contains 20 ml of cold dissection buffer. (Dissect 2 pups for 2 x 106 cells/flask). 2. Carefully pour tissue into a 10 cm dish and gently mince tissue with sterile scissors or razor blade. 3. Transfer tissue to back to 50 ml tube and add 5 ml 1X trypsin and 50 uL DNAse for 25 min at 37ºC. Swirl tube every 5 min. 4. Wash the cortices with Glial Medium twice. 5. Dissociate the tissue by gently triturating the cortices through a 5 ml or 2 ml pipette, followed by a fire-polished Pasteur pipette (3 X 3 triturations). Each time fill pipette with dissociated cells and transfer supernatant to a fresh tube. 6. Dilute cell suspension to 10 ml of Glial Medium, and pass through a 40 uM cell strainer. 7. Spin down the cells at 1700 rpm for 5 min. 8. Re-suspend the cells with 10 ml of Glial Medium, and count. 9. Seed 2 x 106 cells/flask in 15 ml Glial medium. ****(2.0 x 106 cells/flask = 1.33 x 105 cells/ml = 2.67 x 104 cells/cm2)***** 10. Change the medium each of the next two days by aspirating the medium, and then adding back 15 ml of fresh Glial Medium. -
Neuroglial Response to Neuron Injury. a Study Using Intraneural Injection of Ricinus Communis Agglutinin-60
J. Anat. (1989), 164, pp. 201-213 201 With 16 figures Printed in Great Britain Neuroglial response to neuron injury. A study using intraneural injection of ricinus communis agglutinin-60 E. A. LING, C. Y. WEN*, J. Y. SHIEH*, T. Y. YICK AND S. K. LEONG Department of Anatomy, Faculty of Medicine, National University of Singapore, Singapore 0511 and * Department of Anatomy, College of Medicine, National Taiwan University, Taipei, Taiwan 10018 (Accepted 27 September 1988) INTRODUCTION Several studies have shown that ricinus communis agglutinin (RCA), when injected into a nerve in minute amounts, is retrogradely transported by axons in the nerve, resulting in a selective destruction of the parental cell bodies. Thus, the administration of RCA into the vagus nerve would cause a selective destruction of the efferent neurons in the dorsal motor nucleus (Wiley, Blessing & Reis, 1982; Ling & Leong, 1987, 1988). Such a 'suicide transport' of the toxic lectin is also evident in sensory neurons (Yamamoto, Iwasaki & Konno, 1983, 1984; Johnson, Westrum, Henry & Canfield, 1985; Wiley & Oeltmann, 1986). Recently, the use of RCA has become increasingly important as a research tool for tracing the central projections of primary afferents of peripheral nerves (Yamamoto et al. 1983; Leong & Tan, 1987; Ling & Leong, 1987). While much is known about the consequent death of neurons following RCA application, little is known about the response of the non-neuronal cells either closely associated with, or in the vicinity of, the degenerating neurons. According to Yamamoto et al. (1984), the selective destruction of neurons in the trigeminal and dorsal root ganglia by RCA could possibly stimulate the capsule cells involved in the phagocytosis of the degenerating nerve cells. -
University International
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The Functional Organization of Descending Sensory-Motor 2 Pathways in Drosophila 3 4 5 Shigehiro Namiki,1 Michael H
bioRxiv preprint doi: https://doi.org/10.1101/231696; this version posted December 11, 2017. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 1 The functional organization of descending sensory-motor 2 pathways in Drosophila 3 4 5 Shigehiro Namiki,1 Michael H. Dickinson,2 Allan M. Wong,1 Wyatt Korff,1 Gwyneth M. 6 Card,1,* 7 8 1Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA 9 2Division of Biology and Bioengineering, California Institute of Technology, Pasadena, CA 91125, USA 10 11 12 This manuscript includes 55 pages of typescript, 15 figures, 0 tables, 24 supplemental figures, and 6 13 supplementary tables. 14 15 KEYWORDS: command neuron; descending neuron; motor control; neuron database; ventral 16 nervous system 17 18 *Correspondence should be addressed to [email protected] (G.M.C) 19 1 bioRxiv preprint doi: https://doi.org/10.1101/231696; this version posted December 11, 2017. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 20 SUMMARY 21 22 In most animals, the brain controls the body via a set of descending neurons (DNs) that traverse the neck 23 and terminate in post-cranial regions of the nervous system. This critical neural population is thought to 24 activate, maintain and modulate locomotion and other behaviors. Although individual members of this 25 cell class have been well-studied across species ranging from insects to primates, little is known about the 26 overall connectivity pattern of DNs as a population. -
Regulation of Myelin Structure and Conduction Velocity by Perinodal Astrocytes
Correction NEUROSCIENCE Correction for “Regulation of myelin structure and conduc- tion velocity by perinodal astrocytes,” by Dipankar J. Dutta, Dong Ho Woo, Philip R. Lee, Sinisa Pajevic, Olena Bukalo, William C. Huffman, Hiroaki Wake, Peter J. Basser, Shahriar SheikhBahaei, Vanja Lazarevic, Jeffrey C. Smith, and R. Douglas Fields, which was first published October 29, 2018; 10.1073/ pnas.1811013115 (Proc. Natl. Acad. Sci. U.S.A. 115,11832–11837). The authors note that the following statement should be added to the Acknowledgments: “We acknowledge Dr. Hae Ung Lee for preliminary experiments that informed the ultimate experimental approach.” Published under the PNAS license. Published online June 10, 2019. www.pnas.org/cgi/doi/10.1073/pnas.1908361116 12574 | PNAS | June 18, 2019 | vol. 116 | no. 25 www.pnas.org Downloaded by guest on October 2, 2021 Regulation of myelin structure and conduction velocity by perinodal astrocytes Dipankar J. Duttaa,b, Dong Ho Wooa, Philip R. Leea, Sinisa Pajevicc, Olena Bukaloa, William C. Huffmana, Hiroaki Wakea, Peter J. Basserd, Shahriar SheikhBahaeie, Vanja Lazarevicf, Jeffrey C. Smithe, and R. Douglas Fieldsa,1 aSection on Nervous System Development and Plasticity, The Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892; bThe Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc., Bethesda, MD 20817; cMathematical and Statistical Computing Laboratory, Office of Intramural Research, Center for Information -
Structural and Developmental Principles of Neuropil Assembly in C
Article Structural and developmental principles of neuropil assembly in C. elegans https://doi.org/10.1038/s41586-020-03169-5 Mark W. Moyle1, Kristopher M. Barnes2, Manik Kuchroo3, Alex Gonopolskiy3, Leighton H. Duncan1, Titas Sengupta1, Lin Shao1, Min Guo4, Anthony Santella2, Received: 21 April 2020 Ryan Christensen4, Abhishek Kumar5, Yicong Wu4, Kevin R. Moon6, Guy Wolf7, Accepted: 12 November 2020 Smita Krishnaswamy3,10, Zhirong Bao2,10, Hari Shroff4,5,10, William A. Mohler8,10 & Daniel A. Colón-Ramos1,5,9,10 ✉ Published online: xx xx xxxx Check for updates Neuropil is a fundamental form of tissue organization within the brain1, in which densely packed neurons synaptically interconnect into precise circuit architecture2,3. However, the structural and developmental principles that govern this nanoscale precision remain largely unknown4,5. Here we use an iterative data coarse-graining algorithm termed ‘difusion condensation’6 to identify nested circuit structures within the Caenorhabditis elegans neuropil, which is known as the nerve ring. We show that the nerve ring neuropil is largely organized into four strata that are composed of related behavioural circuits. The stratifed architecture of the neuropil is a geometrical representation of the functional segregation of sensory information and motor outputs, with specifc sensory organs and muscle quadrants mapping onto particular neuropil strata. We identify groups of neurons with unique morphologies that integrate information across strata and that create neural structures that cage the strata within the nerve ring. We use high resolution light-sheet microscopy7,8 coupled with lineage-tracing and cell-tracking algorithms9,10 to resolve the developmental sequence and reveal principles of cell position, migration and outgrowth that guide stratifed neuropil organization. -
Oligodendrocyte Precursor Cells Sculpt the Visual System by Regulating Axonal Remodeling
bioRxiv preprint doi: https://doi.org/10.1101/2021.03.11.434829; this version posted March 12, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 1 Oligodendrocyte Precursor Cells Sculpt the Visual System by Regulating 2 Axonal Remodeling 3 4 Yan Xiao1, Laura J Hoodless2, Luigi Petrucco3,4, Ruben Portugues2,4,5, Tim Czopka1,2,5 5 1) Institute of Neuronal Cell Biology, Technical University of Munich, Germany 6 2) Centre for Clinical Brain Sciences, University of Edinburgh, United Kingdom 7 3) Max Planck Institute of Neurobiology, Sensorimotor Control Research Group, Martinsried 82152, 8 Germany 9 4) Institute of Neuroscience, Technical University of Munich, Germany 10 5) Munich Cluster for Systems Neurology (SyNergy), Germany 11 12 13 *Correspondence to 14 15 Dr. Tim Czopka 16 University of Edinburgh 17 Centre for Clinical Brain Sciences 18 Chancellor's Building 19 49 Little France Crescent 20 Edinburgh EH16 4SB, United Kingdom 21 [email protected] bioRxiv preprint doi: https://doi.org/10.1101/2021.03.11.434829; this version posted March 12, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 22 Abstract 23 Many oligodendrocyte precursor cells (OPCs) do not differentiate to form myelin, suggesting 24 additional roles of this cell population. -
Characterization of Oligodendrocyte Lineage Precursor Cells in the Mouse Cerebral Cortex: a Confocal Microscopy Approach to Demyelinating Diseases
IJAE Vol. 115, n. 1/2: 95-102, 2010 ITALIAN JOURNAL OF ANATOMY AND EMBRYOLOGY Characterization of oligodendrocyte lineage precursor cells in the mouse cerebral cortex: a confocal microscopy approach to demyelinating diseases Francesco Girolamo*, Maurizio Strippoli, Mariella Errede, Vincenzo Benagiano, Luisa Roncali, Glauco Ambrosi, Daniela Virgintino Dipartimento di Anatomia Umana e di Istologia ‘Rodolfo Amprino’, Facoltà di Medicina e Chirurgia – Policlinico – Università di Bari, Italia. *Corresponding author, Email: [email protected] Presented at a meeting in honour of Prof. G. Orlandini, Florence, February 15, 2010 Summary The identifi cation of stem cells resident in the adult central nervous system has redirected the focus of research into demyelinating diseases, such as multiple sclerosis, mainly affecting the brain white matter. This immunocytochemical and morphometrical study was carried out by confocal microscopy in the adult mouse cerebral cortex, with the aim of analysing, in the brain grey matter, the characteristics of the oligodendrocyte lineage cells, whose capability to remyeli nate is still controversial. The observations demonstrated the presence in all the cortex layers of glial restricted progenitors, reactive to A2B5 marker, oligodendrocyte precursor cells, expressing the NG2 proteoglycan, and preoligodendrocytes and premyelinating oligodendrocytes, reac tive to the specifi c marker O4. NG2 expressing cells constitute the major immature population of the cortex, since not only oligodendrocyte precursor cells and preoligodendrocytes but also a part of the glial restrict progenitors express the NG2 proteoglycan. Together with the popula tion of these immature cells, a larger population of mature oligodendrocytes was revealed by the classical oligodendrocyte and myelin markers, 2’,3’cyclic nucleotide 3’phosphodiesterase, myelin basic protein and myelin oligodendrocyte glycoprotein. -
Microglia Activation Triggers Astrocyte-Mediated Modulation of Excitatory Neurotransmission
Microglia activation triggers astrocyte-mediated PNAS PLUS modulation of excitatory neurotransmission Olivier Pascuala,b,c,1,2, Sarrah Ben Achoura,b,c,2, Philippe Rostainga,b,c, Antoine Trillera,b,c, and Alain Bessisa,b,c aInstitut de Biologie de l’Ecole Normale Supérieure, F-75005 Paris, France; bInstitut National de la Santé et de la Recherche Médicale U1024, F-75005 Paris, France; and cCentre National de la Recherche Scientifique, Unité Mixte de Recherche 8197, F-75005 Paris, France Edited* by Tullio Pozzan, University of Padova, Padua, Italy, and approved November 21, 2011 (received for review July 18, 2011) Fine control of neuronal activity is crucial to rapidly adjust to subtle tatively able to sense neuronal activity and/or communicate with changes of the environment. This fine tuning was thought to be astrocytes. In response to stimuli, microglia are activated, and they purely neuronal until the discovery that astrocytes are active players release neurotransmitters (19), which are small molecules such as of synaptic transmission. In the adult hippocampus, microglia are nitric oxide, trophic factors, or cytokines, all known to control the other major glial cell type. Microglia are highly dynamic and neuronal function and synaptic transmission (20, 21). In addition, closely associated with neurons and astrocytes. They react rapidly to changes in plasticity and neuronal activity have been shown to modifications of their environment and are able to release mole- modify the resident time of microglia processes at synapses (22). cules known to control neuronal function and synaptic transmission. Although long-term effects of microglial activation and in- Therefore, microglia display functional features of synaptic part- flammation have been studied (14, 23, 24), early consequences of ners, but their involvement in the regulation of synaptic trans- such activation are still unknown, especially the cell type involved mission has not yet been addressed. -
Astrocyte Failure As a Cause of CNS Dysfunction
Molecular Psychiatry (2000) 5, 230–232 2000 Macmillan Publishers Ltd All rights reserved 1359-4184/00 $15.00 www.nature.com/mp NEWS & VIEWS Astrocyte failure as a cause of CNS dysfunction All insults to the central nervous systems (CNS), expressing HSV-Tk from the mouse Gfap promoter, including injury, ischemia, infection and degenerative reactive, transgene-expressing astrocytes adjacent to a disease are invariably accompanied by the hypertro- forebrain stab injury are ablated by GCV.8,9 These and phy, altered gene expression and proliferation of astro- other studies have demonstrated the essential nature of cytes, a process commonly referred to as ‘reactive astrocyte functions in a number of contexts related to astrocytosis’. While much is known about molecules the response to injury, and highlighted how astrocyte that either influence, or are produced by, reactive astro- failure might lead to CNS dysfunction in various ways. cytes,1,2 the functions of these cells are incompletely understood. Astrocytes are the most numerous cells in Astrocytes, the blood–brain barrier and interstitial the vertebrate central nervous system (CNS), and vari- edema ous functions have been ascribed to them in the unin- jured CNS, including: provision of structural support The anatomical correlate of the BBB is thought to for neural elements (neuro-glia = neural ‘glue’); homeo- reside in tight junctions between endothelial cells of static maintenance of the extracellular ionic environ- cerebral capillaries, which are of high electrical resist- ment and pH;