Abstract
During brain development, radial glial (RG) cells and the different progenitor subtypes are characterized by their bipolar morphology that includes an ovoid cell body and one or two radial processes that span across the developing cerebral wall. Different cells transport the reduced form of vitamin C, ascorbic acid (AA), using sodium-dependent ascorbic acid cotransporters (SVCT1 or SVCT2). SVCT2 is mainly expressed in the nervous system (CNS); however, its localization in the central nervous system during embryonic development along with the mechanism by which RG take up vitamin C and its intracellular effects is unknown. Thus, we sought to determine the expression and localization of SVCT2 during CNS development. SVCT2 is preferentially localized in the RG body at the ventricular edge of the cortex during the neurogenic stage (E12 to E17). The localization of SVCT2 overexpressed by in utero electroporation of E14 embryos is consistent with ventricular polarization. A similar distribution pattern was observed in human brain tissue sections at 9 weeks of gestation; however, SVCT2 immunoreaction was also detected in the inner and outer subventricular zone (SVZ). Finally, we used C17.2 neural stem cell line, J1ES cells and primary cell cultures derived from the brain cortex to analyze functional SVCT2 activity, AA effects in progenitor cells bipolar morphology, and SVCT2 expression levels in different culture conditions. Our results indicate that basal RG cells and apical intermediate and subapical progenitors are the main cell types expressing SVCT2 in the lissencephalic brain. SVCT2 was mainly detected in the apical region of the ventricular zone cells, contacting the cerebrospinal fluid. In gyrencephalic brains, SVCT2 was also detected in progenitor cells located in the inner and outer SVZ. Finally, we defined that AA has a strong radializing (bipolar morphology) effect in progenitor cells in culture and the differentiation condition modulates SVCT2 expression.
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Abbreviations
- AA:
-
Ascorbic acid
- DHA:
-
Dehydroascorbic acid
- DMEM:
-
Dulbecco’s modified Eagle medium
- FBS:
-
Fetal bovine serum
- DOG:
-
deoxyglucose
- GFAP:
-
Glial fibrillary acidic protein
- GLUT:
-
Glucose transporter
- IMDM:
-
Iscove’s Modified Dulbecco’s medium
- NSC:
-
Neural stem cell
- PAGE:
-
Polyacrylamide gel electrophoresis
- PBS:
-
Phosphate–buffered saline
- RG:
-
Radial glia
- SVCT:
-
Sodium-dependent ascorbic acid cotransporters
- SDS:
-
Sodium dodecyl sulfate
- VZ:
-
Ventricular zone.
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Acknowledgments
This work was supported by grants from the Fondo Nacional de Ciencia y Tecnología (FONDECYT 1140477 FN, 11130529 to CSA, 11140405 to KS, 11150678 to FM) and Conicyt-PIA CMA BIO BIO ECM-12 to FN. We thank Dra. Marjet Heitzer for critically reading and editing the manuscript and Ms. Ximena Koch for the technical support.
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All animals were handled in strict accordance with the Animal Welfare Assurance (permit number 2010101A), and all animal work was approved by the appropriate Ethics and Animal Care and Use Committee of the University of Concepcion, Chile.
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Silva-Álvarez, C., Salazar, K., Cisternas, P. et al. Apical Polarization of SVCT2 in Apical Radial Glial Cells and Progenitors During Brain Development. Mol Neurobiol 54, 5449–5467 (2017). https://doi.org/10.1007/s12035-016-0081-2
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DOI: https://doi.org/10.1007/s12035-016-0081-2