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
ATP Binding Cassette Transporter, Subfamily G, Member 2
ATP-Binding Cassette Transporters/biosynthesis
Aldehyde Dehydrogenase/metabolism
Animals
Antigens, Differentiation/biosynthesis
Carrier Proteins/biosynthesis
Cell Communication/physiology
Cell Count
Cell Differentiation/genetics,physiology
Cell Membrane/chemistry,metabolism
Cells, Cultured
Connexin 43/genetics,metabolism
Connexins/genetics,metabolism
DNA-Binding Proteins/biosynthesis
Electrophysiology
GTP-Binding Proteins
Gap Junctions/physiology
Gene Expression Profiling
Glucose Transporter Type 1
High Mobility Group Proteins/biosynthesis
Immunohistochemistry
Ion Channels/biosynthesis,genetics
Monosaccharide Transport Proteins/biosynthesis,genetics
Multipotent Stem Cells/cytology,drug effects,metabolism
Neoplasm Proteins/biosynthesis
Neurons/cytology
Nuclear Proteins/biosynthesis
Oligonucleotide Array Sequence Analysis
Patch-Clamp Techniques
Rats
Rats, Sprague-Dawley
Receptors, Muscarinic/drug effects,metabolism
Receptors, Nicotinic/drug effects,metabolism
SOXB1 Transcription Factors
Signal Transduction/genetics,physiology
Chemicals
ABCG2 protein, human
ATP Binding Cassette Transporter, Subfamily G, Member 2
ATP-Binding Cassette Transporters
Antigens, Differentiation
Carrier Proteins
Connexin 43
Connexins
DNA-Binding Proteins
Glucose Transporter Type 1
Gnl3 protein, rat
High Mobility Group Proteins
Ion Channels
Monosaccharide Transport Proteins
Neoplasm Proteins
Nuclear Proteins
Receptors, Muscarinic
Receptors, Nicotinic
SOX1 protein, human
SOXB1 Transcription Factors
Slc2a1 protein, rat
Sox1 protein, rat
connexin 45
Aldehyde Dehydrogenase
GTP-Binding Proteins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Cai Jingli
Laboratory of Neurosciences, Gerontology Research Center, National Institute on Aging, Baltimore, Maryland 21224, USA.
Cheng Aiwu
Luo Yongquan
Lu Chengbiao
Mattson Mark P
Rao Mahendra S
Furukawa Katsutoshi