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Author Van Liefferinge, Joeri ♦ Bentea, Eduard ♦ Demuyser, Thomas ♦ Albertini, Giulia ♦ Follin-Arbelet, Virginie ♦ Holmseth, Silvia ♦ Merckx, Ellen ♦ Sato, Hideyo ♦ Aerts, Joeri L. ♦ Smolders, Ilse ♦ Arckens, Lutgarde ♦ Danbolt, Niels C. ♦ Massie, Ann
Source World Health Organization (WHO)-Global Index Medicus
Content type Text
Publisher Wiley
File Format HTM / HTML
Language English
Difficulty Level Medium
Subject Domain (in DDC) Computer science, information & general works ♦ Library & information sciences ♦ Natural sciences & mathematics ♦ Chemistry & allied sciences ♦ Life sciences; biology ♦ Physiology & related subjects ♦ Biochemistry ♦ Natural history of organisms ♦ Technology ♦ Medicine & health ♦ Human anatomy, cytology, histology ♦ Human physiology ♦ Pharmacology and therapeutics ♦ Diseases ♦ Manufacture for specific uses ♦ Precision instruments & other devices
Subject Domain (in MeSH) Nervous System ♦ Anatomy ♦ Eukaryota ♦ Organisms ♦ Amino Acids, Peptides, and Proteins ♦ Chemicals and Drugs ♦ Investigative Techniques ♦ Analytical, Diagnostic and Therapeutic Techniques and Equipment ♦ Chemical Phenomena ♦ Biological Sciences ♦ Information Science ♦ Information Science
Subject Keyword Discipline Neurology ♦ Amino Acid Transport System Y+ ♦ Biosynthesis ♦ Genetics ♦ Autoantibodies ♦ Metabolism ♦ Brain ♦ Amino Acid Sequence ♦ Animals ♦ Humans ♦ Male ♦ Mice ♦ Mice, Knockout ♦ Molecular Sequence Data ♦ Rabbits ♦ Rats ♦ Sequence Analysis, Protein ♦ Methods ♦ Sequence Homology ♦ Comparative Study ♦ Journal Article ♦ Research Support, Non-u.s. Gov't
Abstract The cystine/glutamate antiporter or system Xc- exchanges cystine for glutamate, thereby supporting intracellular glutathione synthesis and nonvesicular glutamate release. The role of system Xc- in neurological disorders can be dual and remains a matter of debate. One important reason for the contradictory findings that have been reported to date is the use of nonspecific anti-xCT (the specific subunit of system Xc-) antibodies. Often studies rely on the predicted molecular weight of 55.5 kDa to identify xCT on Western blots. However, using brain extracts from xCT knockout (xCT(-/-)) mice as negative controls, we show that xCT migrates as a 35-kDa protein. Misinterpretation of immunoblots leads to incorrect assessment of antibody specificity and thereby to erroneous data interpretation. Here we have verified the specificity of most commonly used commercial and some in-house-developed anti-xCT antibodies by comparing their immunoreactivity in brain tissue of xCT(+/+) and xCT(-/-) mice by Western blotting and immunohistochemistry. The Western blot screening results demonstrate that antibody specificity not only differs between batches produced by immunizing different rabbits with the same antigen but also between bleedings of the same rabbit. Moreover, distinct immunohistochemical protocols have been tested for all the anti-xCT antibodies that were specific on Western blots in order to obtain a specific immunolabeling. Only one of our in-house-developed antibodies could reveal specific xCT labeling and exclusively on acetone-postfixed cryosections. Using this approach, we observed xCT protein expression throughout the mouse forebrain, including cortex, striatum, hippocampus, midbrain, thalamus, and amygdala, with greatest expression in regions facing the cerebrospinal fluid and meninges.
Description Country affiliation: Belgium
Author Affiliation: Van Liefferinge J ( Department of Pharmaceutical Chemistry and Drug Analysis, Center for Neurosciences, Vrije Universiteit Brussel, Brussels, 1090, Belgium.); Bentea E ( Department of Pharmaceutical Biotechnology and Molecular Biology, Center for Neurosciences, Vrije Universiteit Brussel, Brussels, 1090, Belgium.); Demuyser T ( Department of Pharmaceutical Chemistry and Drug Analysis, Center for Neurosciences, Vrije Universiteit Brussel, Brussels, 1090, Belgium.); Albertini G ( Department of Pharmaceutical Chemistry and Drug Analysis, Center for Neurosciences, Vrije Universiteit Brussel, Brussels, 1090, Belgium.); Follin-Arbelet V ( Department of Molecular Medicine, Institute of Basic Medical Sciences, University of Oslo, Oslo, 0317, Norway.); Holmseth S ( Department of Molecular Medicine, Institute of Basic Medical Sciences, University of Oslo, Oslo, 0317, Norway.); Merckx E ( Department of Pharmaceutical Biotechnology and Molecular Biology, Center for Neurosciences, Vrije Universiteit Brussel, Brussels, 1090, Belgium.); Sato H ( Laboratory of Biochemistry and Molecular Biology, Department of Medical Technology, Niigata University, Niigata, Niigata Prefecture, 950-2181, Japan.); Aerts JL ( Laboratory of Molecular and Cellular Therapy, Department of Immunology-Physiology, Vrije Universiteit Brussel, Brussels, 1090, Belgium.); Smolders I ( Department of Pharmaceutical Chemistry and Drug Analysis, Center for Neurosciences, Vrije Universiteit Brussel, Brussels, 1090, Belgium.); Arckens L ( Laboratory of Neuroplasticity and Neuroproteomics, KU Leuven, Leuven, 3000, Belgium.); Danbolt NC ( Department of Molecular Medicine, Institute of Basic Medical Sciences, University of Oslo, Oslo, 0317, Norway.); Massie A ( Department of Pharmaceutical Biotechnology and Molecular Biology, Center for Neurosciences, Vrije Universiteit Brussel, Brussels, 1090, Belgium.)
ISSN 00219967
Educational Role Student ♦ Teacher
Age Range above 22 year
Educational Use Reading ♦ Research ♦ Self Learning
Interactivity Type Expositive
Education Level UG and PG
Learning Resource Type Article
Publisher Date 2016-04-01
Publisher Place United States
e-ISSN 10969861
Journal Journal of Comparative Neurology
Volume Number 524
Issue Number 5


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Source: WHO-Global Index Medicus