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Please use this identifier to cite or link to this item: http://hdl.handle.net/10481/37092

Title: Expression of Glucose Transporters in the Prelaminar Region of the Optic-Nerve Head of the Pig as Determined by Immunolabeling and Tissue Culture
Authors: Carreras Egaña, Francisco Javier
Aranda, Carlos J.
Porcel, David
Rodríguez-Hurtado, Francisco
Martínez-Agustín, Olga
Zarzuelo Zurita, Antonio
Issue Date: 2015
Abstract: Background: To develop the use of cultured tissue of the prelaminar optic nerve of the pig to explore possible alterations of the astrocyte-axon metabolic pathways in glaucoma, we map the distribution of the glucose transporters GLUT1 and GLUT3 in fresh and cultured tissue.
Methods: We monitor cell survival in cultures of the prelaminar optic-nerve tissue, measuring necrosis and apoptosis markers biochemically as well as morphologically, and establish the presence of the glucose transporters GLUT1 and GLUT3. We map the distribution of these transporters with immunolabeling in histological sections of the optic nerve using confocal and electronic transmission microscopy.
Results: We find that the main death type in prelaminar culture is apoptosis. Caspase 7 staining reveals an increment in apoptosis from day 1 to day 4 and a reduction from day 4 to day 8. Western blotting for GLUT1 shows stability with increased culture time. CLSM micrographs locate GLUT1 in the columnar astrocytes and in the area of axonal bundles. Anti-GLUT3 predominantly labels axonal bundles. TEM immunolabeling with colloidal gold displays a very specific distribution of GLUT-1 in the membranes of vascular endothelial cells and in periaxonal astrocyte expansions. The GLUT-3 isoform is observed with TEM only in axons in the axonal bundles.
Conclusions: Tissue culture is suitable for apoptosis-induction experiments. The results suggest that glucose is transported to the axonal cleft intracytoplasmically and delivered to the cleft by GLUT1 transporters. As monocarboxylate transporters have been reported in the prelaminar region of the optic-nerve head, this area is likely to use both lactate and glucose as energy sources.
Sponsorship: This work was supported by a grant from the Consejería de Salud, Junta de Andalucia, Spain, Project PI-0655-2013.
Publisher: Public Library of Science (Plos)
Keywords: Astrocytes
Apoptosis
Glucose
Axons
Confocal laser microscopy
Transmission electron microscopy
DAPI staining
Glucose metabolism
URI: http://hdl.handle.net/10481/37092
Rights : Creative Commons Attribution-NonCommercial-NoDerivs 3.0 License
Citation: Carreras, F.J.; et al. Expression of Glucose Transporters in the Prelaminar Region of the Optic-Nerve Head of the Pig as Determined by Immunolabeling and Tissue Culture. Plos One, 10(6): e0128516 (2015). [http://hdl.handle.net/10481/37092]
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