Expression of Metabolic Coupling and Adhesion Proteins in the Porcine Optic-Nerve Head: Relevance to a Flow Model of Glaucoma

Carreras, Francisco-Javier; Porcel, David; Rodríguez-Hurtado, Francisco Jorge; Zarzuelo, Antonio; Carreras, Ignacio; Galisteo, Milagros

doi:10.5772/20546

Cited by 3 publications

(7 citation statements)

References 46 publications

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“…[ 18 , 19 ]. The distribution of MCT1 and 2 isoforms in the prelaminar region has recently been described as exclusively around the axonal bundles [ 20 ], confirming the hypothesis that considers the lactate shuttle to be an important source of axonal energy in the preliminary tissue of the ONH.…”

Section: Introductionsupporting

confidence: 53%

“…As reported elsewhere, axons are destroyed, leaving a matrix of astrocytes and capillaries that adapt to culture conditions. [ 20 ] A comparison between the freshly excised and the cultured tissues by microscopic and analytical methods renders visible the distribution and proportional quantities of the coupled molecules under study. This approach was used in a previous report with MCTs and is performed here with GLUTs.…”

Section: Methodsmentioning

confidence: 99%

“…This approach was used in a previous report with MCTs and is performed here with GLUTs. [ 20 ] Fig 2A gives a general idea of the tissue studied to express the main doubt that the work seeks to elucidate, as stated in the Introduction, and to stress the difficulties inherent in an intricate tissue organization that justify the following combination of different approaches: confocal laser scanning microscopy (CLSM), transmission electron microscopy (TEM) and western blotting of both fresh and cultured specimens. Any single approach to addressing the presence and distribution of GLUT molecules in the prelaminar tissue of the ONH would be insufficient on its own.…”

Section: Methodsmentioning

confidence: 99%

“…Explants, 100 to 300 μm thick, of the dissected prelaminar region of the ONH of recently enucleated pig eyes were cultured in vitro following the method described by Stoppini et al (1991) [ 29 ], as adapted by Carreras et al (2011) [ 20 ]. Briefly, the optic discs were dissected with a surgical blade in a sterile environment.…”

Section: Methodsmentioning

confidence: 99%

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Expression of Glucose Transporters in the Prelaminar Region of the Optic-Nerve Head of the Pig as Determined by Immunolabeling and Tissue Culture

et al. 2015

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BackgroundTo 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.MethodsWe 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.ResultsWe 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.ConclusionsTissue 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.

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Section: Introductionsupporting

confidence: 53%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Expression of Glucose Transporters in the Prelaminar Region of the Optic-Nerve Head of the Pig as Determined by Immunolabeling and Tissue Culture

et al. 2015

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“…Lactate is then transported into the axons through monocarboxylate transporters (MCT) [38]. Gap junctions, also present in the prelaminar tissue [39], allow astrocytes to share glucose and lactate. Critical to the maintenance of correct cellular relationships is the presence of adherens junctions rich in N-Cadherine, a calcium-dependent adhesion molecule.…”

Section: A New Member Of the Metabolic Paradigm?mentioning

confidence: 99%

Glaucoma as a Tax on Hominid Evolution. Constraints of the Experimental Animal Models of Glaucoma

Carreras¹

2016

J Clin Exp Ophthalmol

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Glaucoma continues to be a mysterious disease. High intraocular pressure (IOP), once the landmark of the disease, has been relegated to the humble role of risk factor, despite the fact that lowering IOP continues to be the only partially successful treatment. Even with an IOP successfully controlled to statistical standards, many patients still mysteriously continue to progressively lose neural tissue. A novel pathogenetical mechanism has recently been put forward as the main agent of the disease. In this new light, some structural details of the human eye, the result of the peculiar evolutionary development, take on an unexpected role in the presentation of the disease. As results of cladistics analysis, the human eye appears as particularly prone to suffering glaucoma as a consequence of the evolutionary path taken. That makes glaucoma essentially a human (and related hominid) scourge. Exclusive features of the human eye compared to other vertebrates involve both the anterior and posterior segment. Those features that facilitate glaucoma in humans are absent in many of the purported animal models for the disease, and this inadequacy further convolutes the mystery of glaucoma. This suggests the division of the animal models into those that mimic the whole disease and those that only reproduce a pertinent histological feature. Unfortunately most animal models are based on high intraocular pressure which excludes most cases of low or moderate pressure. For any animal model, it is important the researcher accurately establishes the constraints of the model in order to avoid jumping to conclusions.

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Virtual Tissue Engineering and Optic Pathways: Plotting the Course of the Axons in the Retinal Nerve Fiber Layer

Carreras

Medina

Ruiz-Lozano

et al. 2014

Invest. Ophthalmol. Vis. Sci.

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Expression of Metabolic Coupling and Adhesion Proteins in the Porcine Optic-Nerve Head: Relevance to a Flow Model of Glaucoma

Cited by 3 publications

References 46 publications

Expression of Glucose Transporters in the Prelaminar Region of the Optic-Nerve Head of the Pig as Determined by Immunolabeling and Tissue Culture

Expression of Glucose Transporters in the Prelaminar Region of the Optic-Nerve Head of the Pig as Determined by Immunolabeling and Tissue Culture

Glaucoma as a Tax on Hominid Evolution. Constraints of the Experimental Animal Models of Glaucoma

Virtual Tissue Engineering and Optic Pathways: Plotting the Course of the Axons in the Retinal Nerve Fiber Layer

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