Primary cultures of astrocytes and neurons are frequently used to investigate metabolic properties of these two important brain cell types. Here we describe methods to generate primary cultures that are highly enriched in astrocytes or cerebellar granule neurons. These cultures are good model systems to investigate the basal metabolism of astrocytes and neurons as well as metabolite export from these cells. As examples for such studies, we describe here in detail the robust practical procedures that we use to investigate cellular glucose consumption and lactate production as well as the cellular contents and the export of glutathione from cultured brain cells. In this context we also describe some viability assays that are useful to confi rm that a given experimental treatment is not toxic for cultured neural cells.

scihub/10.1007/978-1-4939-1059-5_3.pdf

Brain Energy Metabolism (Neuromethods, 90)

edited by Johannes Hirrlinger, Carl-Ludwig-Institute for Physiology, University of Leipzig, Leipzig, Germany, Helle S. Waagepetersen, Faculty of health and Medical Sciences, University of Copenhagen, Denmark

Heidelberg; Humana Press; Humana

Heidelberg : Springer

Neuromethods -- v. 90, Springer Protocols, Neuromethods -- v. 90., Springer protocols (Series), New York, New York State, 2014

United States, United States of America

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Referenced by: doi:10.1016/j.brainresrev.2009.10.003 doi:10.1007/978-1-4614-1788-0_36 doi:10.1111/j.1471-4159.2012.07664.x doi:10.1007/s11064-012-0868-0 doi:10.1016/0076-6879(85)09097-8 doi:10.1007/978-1-59745-261-8_25 doi:10.1016/s1385-299x(97)00047-0 doi:10.1016/j.neuint.2010.06.010 doi:10.1046/j.1471-4159.1999.0731422.x doi:10.1002/glia.20901 doi:10.1016/s0021-9258(19)52451-6 doi:10.1007/bf00297072 doi:10.1080/10715760400023671 doi:10.1073/pnas.62.3.813 doi:10.1111/j.1471-4159.1992.tb09750.x doi:10.1002/jnr.22093 doi:10.1016/0006-8993(93)91429-v doi:10.3389/neuro.14.001.2009 doi:10.1016/0003-2697(69)90064-5 doi:10.1046/j.1471-4159.1996.67041375.x doi:10.1023/a:1027310328310 doi:10.1016/s0076-6879(05)00023-6 doi:10.1016/0003-2697(80)90139-6 doi:10.1016/s0304-3940(00)01369-0 doi:10.1046/j.1471-4159.2003.02235.x doi:10.1177/1073858403254006 doi:10.1002/jnr.22638 doi:10.1007/s11064-012-0760-y doi:10.1046/j.1471-4159.1995.65020590.x doi:10.1046/j.1471-4159.2003.01871.x doi:10.1046/j.1471-4159.2002.00999.x doi:10.1515/hsz-2012-0145

Includes bibliographical references and index.

Determination of CO2 Production in Subcellular Preparations like Synaptosomes and Isolated Mitochondria Using 14C-Labeled Substrates and Radioactive CO2 Measurements
Transport of Lactate: Characterization of the Transporters Involved in Transport at the Plasma Membrane by Heterologous Protein Expression in Xenopus Oocytes
Primary Cultures of Astrocytes and Neurons as Model Systems to Study the Metabolism and Metabolite Export from Brain Cells
Metabolic Mapping of Astrocytes and Neurons in Culture Using Stable Isotopes and Gas Chromatography-Mass Spectrometry (GC-MS)
Metabolic Flux Analysis Tools to Investigate Brain Metabolism In Vitro
Fluorescent Nanosensor Based Flux Analysis: Overview and the Example of Glucose
Mitochondrial Bioenergetics Assessed by Functional Fluorescence Dyes
RNA Interference as a Tool to Selectively Down-Modulate Protein Function
Localizing and Quantifying Metabolites In Situ with Luminometry: Induced Metabolic Bioluminescence Imaging (imBI)
A Chip Off the Old Block: The Brain Slice as a Model for Metabolic Studies of Brain Compartmentation and Neuropharmacology
Integrated Measurements of Electrical Activity, Oxygen Tension, Blood Flow, and Ca2+-Signaling in Rodents In Vivo
Measuring Cerebral Hemodynamics and Energy Metabolism by Near-Infrared Spectroscopy
Compartmental Analysis of Metabolism by 13C Magnetic Resonance Spectroscopy
Positron Emission Tomography of Brain Glucose Metabolism with [18F]Fluorodeoxyglucose in Humans.

Brain Energy Metabolism addresses its challenging subject by presenting diverse technologies allowing for the investigation of brain energy metabolism on different levels of complexity. Model systems are discussed, starting from the reductionist approach like primary cell cultures which allow assessing of the properties and functions of a single brain cell type with many different types of analysis, however, at the expense of neglecting the interaction between cell types in the brain. On the other end, analysis in animals and humans in vivo is discussed, maintaining the full complexity of the tissue and the organism but making high demands on the methods of analysis. Written for the popular Neuromethods series, chapters include the kind of detailed description and key implementation advice that aims to support reproducible results in the lab.Meticulous and authoritative, Brain Energy Metabolism provides an ideal guide for researchers interested in brain energy metabolism with the hope of stimulating more research in this exciting and very important field.
Erscheinungsdatum: 16.09.2014

Brain Energy Metabolism addresses its challenging subject by presenting diverse technologies allowing for the investigation of brain energy metabolism on different levels of complexity. Model systems are discussed, starting from the reductionist approach like primary cell cultures which allow assessing of the properties and functions of a single brain cell type with many different types of analysis, however, at the expense of neglecting the interaction between cell types in the brain. On the other end, analysis in animals and humans in vivo is discussed, maintaining the full complexity of the tissue and the organism but making high demands on the methods of analysis. Written for the popular Neuromethods series, chapters include the kind of detailed description and key implementation advice that aims to support reproducible results in the lab. Meticulous and authoritative, Brain Energy Metabolism provides an ideal guide for researchers interested in brain energy metabolism with the hope of stimulating more research in this exciting and very important field. --