Abstract
We have characterized several potential stem cell markers and defined the membrane properties of rat fetal (E10.5) neural stem cells (NSC) by immunocytochemistry, electrophysiology and microarray analysis. Immunocytochemical analysis demonstrates specificity of expression of Sox1, ABCG2/Bcrp1, and shows that nucleostemin labels both progenitor and stem cell populations. NSCs, like hematopoietic stem cells, express high levels of aldehyde dehydrogenase (ALDH) as assessed by Aldefluor labeling. Microarray analysis of 96 transporters and channels showed that Glucose transporter 1 (Glut1/Slc2a1) expression is unique to fetal NSCs or other differentiated cells. Electrophysiological examination showed that fetal NSCs respond to acetylcholine and its agonists, such as nicotine and muscarine. NSCs express low levels of tetrodotoxin (TTX) sensitive and insensitive sodium channels and calcium channels while expressing at least three kinds of potassium channels. We find that gap junction communication is mediated by connexin (Cx)43 and Cx45, and is essential for NSC survival and proliferation. Overall, our results show that fetal NSCs exhibit a unique signature that can be used to determine their location and assess their ability to respond to their environment.
MeSH terms
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ATP Binding Cassette Transporter, Subfamily G, Member 2
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ATP-Binding Cassette Transporters / biosynthesis
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Aldehyde Dehydrogenase / metabolism
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Animals
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Antigens, Differentiation / biosynthesis
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Carrier Proteins / biosynthesis
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Cell Communication / physiology
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Cell Count
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Cell Differentiation / genetics
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Cell Differentiation / physiology
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Cell Membrane / chemistry*
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Cell Membrane / metabolism*
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Cells, Cultured
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Connexin 43 / genetics
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Connexin 43 / metabolism
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Connexins / genetics
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Connexins / metabolism
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DNA-Binding Proteins / biosynthesis
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Electrophysiology
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GTP-Binding Proteins
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Gap Junctions / physiology
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Gene Expression Profiling
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Glucose Transporter Type 1
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High Mobility Group Proteins / biosynthesis
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Immunohistochemistry
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Ion Channels / biosynthesis
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Ion Channels / genetics
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Monosaccharide Transport Proteins / biosynthesis
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Monosaccharide Transport Proteins / genetics
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Multipotent Stem Cells / cytology
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Multipotent Stem Cells / drug effects
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Multipotent Stem Cells / metabolism*
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Neoplasm Proteins / biosynthesis
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Neurons* / cytology
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Nuclear Proteins / biosynthesis
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Oligonucleotide Array Sequence Analysis
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Patch-Clamp Techniques
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Rats
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Rats, Sprague-Dawley
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Receptors, Muscarinic / drug effects
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Receptors, Muscarinic / metabolism
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Receptors, Nicotinic / drug effects
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Receptors, Nicotinic / metabolism
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SOXB1 Transcription Factors
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Signal Transduction / genetics
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Signal Transduction / physiology
Substances
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ABCG2 protein, human
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ATP Binding Cassette Transporter, Subfamily G, Member 2
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ATP-Binding Cassette Transporters
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Antigens, Differentiation
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Carrier Proteins
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Connexin 43
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Connexins
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DNA-Binding Proteins
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Glucose Transporter Type 1
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Gnl3 protein, rat
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High Mobility Group Proteins
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Ion Channels
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Monosaccharide Transport Proteins
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Neoplasm Proteins
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Nuclear Proteins
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Receptors, Muscarinic
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Receptors, Nicotinic
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SOX1 protein, human
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SOXB1 Transcription Factors
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Slc2a1 protein, rat
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Sox1 protein, rat
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connexin 45
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Aldehyde Dehydrogenase
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GTP-Binding Proteins