Ocular Hypertension Results in Hypoxia within Glia and Neurons throughout the Visual Projection
<p>OHT results in increased IOP, RGC loss, and functional compromise. (<b>A</b>). Experimental design. Intraperitoneal injection of tamoxifen was administered to 2-month-old CAG-ODD transgenic mice. Visual evoked potential (VEP) and IOP were measured before and after 3, 14, and 28 days of OHT induction. OHT was induced by magnetic bead intracameral injection. Mice received an IP injection of 60 mg/kg pimonidazole, then were sacrificed 90 min later to compare and assess hypoxia using tdTomato (tdT) expression and pimonidazole (pimo) methods at 6 h, 3 days, 14 days, and 28 days after OHT in the retina, ON and visual centers in the brain. (<b>B)</b>. A significant IOP elevation was detected (****, <span class="html-italic">p</span> < 0.0001) 3, 14, and 28 d after OHT. n = 16 eyes at 3 d, 14 eyes at 14 d, 12 eyes at 28 d, and 30 control eyes. (<b>C</b>). A significant decrease (F <sub>(4, 75)</sub> = 17.51) in RGC count per GCL was shown 14 d after OHT compared to control (%, <span class="html-italic">p</span> < 0.0001), 6 h (*, <span class="html-italic">p</span> < 0.0332), and 3 d (&, <span class="html-italic">p</span> = 0.0026). In addition, a significant decrease was detected in RGC count per µm GCL 28d after OHT compared to control (@, <span class="html-italic">p</span> < 0.0001), 6 h (<span>$</span>, <span class="html-italic">p</span> = 0.0052), and 3d (#, <span class="html-italic">p</span> = 0.0002); n = 12 eyes at 6 h, 12 eyes at 3 d, 15 eyes at 14 d, 21 eyes at 28 d, and 20 control eyes. (<b>D</b>). Representative examples of RGC immunofluorescence for the control, 6 h, 3 d, 14 d, and 28 d time points; scale bar = 25 µm. (<b>E</b>). A significant decrease (F <sub>(4, 36)</sub> = 14.64) was shown in axon count 14d after OHT compared to control (%, <span class="html-italic">p</span> < 0.0001), 6 h (*, <span class="html-italic">p</span> = 0.0105), and 3 d (&, <span class="html-italic">p</span> < 0.0425). A significant decrease was also shown in axon count 28 d after OHT compared to control (@, <span class="html-italic">p</span> < 0.0001), 6 h (<span>$</span>, <span class="html-italic">p</span> = 0.0009), and 3 d (#, <span class="html-italic">p</span> = 0.0045); n = 7 ONs at 6 h, 6 ONs at 3 d, 8 ONs at 14 d, 7 ONs at 28 d, and 13 control ONs. Data represented combined results from recombinase negative (cre−) and recombinase positive (cre+) CAG-ODD transgenic mice. Days = d, and hours = h.</p> "> Figure 2
<p>(<b>A</b>). A significant decrease (F <sub>(3, 44)</sub> = 6.119) in N1 amplitude was observed after 3 d OHT (***, <span class="html-italic">p</span> = 0.0010) and 14 d OHT (*, <span class="html-italic">p</span> = 0.0111) compared to control. n = 14 eyes at 3 d, 14 eyes at 14 d, 8 eyes at 28 d, and 12 control eyes. (<b>B</b>). A significant decrease (F <sub>(3, 44)</sub> = 10.60) in P2 amplitude 3 d OHT (****, <span class="html-italic">p</span> < 0.0001), 14 d OHT (*, <span class="html-italic">p</span> = 0.0375), and 28 d OHT (*, <span class="html-italic">p</span> = 0.0490) compared to control in addition to the significant decrease in P2 amplitude 3 d OHT compared to 14 d OHT (*, <span class="html-italic">p</span> = 0.0248). n = 14 eyes at 3 d, 14 eyes at 14 d, 8 eyes at 28 d, and 12 control eyes. (<b>C</b>). A significant decrease (F <sub>(3, 44)</sub> = 14.38) in response amplitude was observed after 3 d OHT (****, <span class="html-italic">p</span> < 0.0001) and 14 d OHT (****, <span class="html-italic">p</span> < 0.0001) compared to control; in addition, the response amplitude at 3 d OHT was significantly lower than that at 28 d OHT (*, <span class="html-italic">p</span> = 0.0181). n = 14 eyes at 3 d, 14 eyes at 14 d, 8 eyes at 28 d, and 12 control eyes. (<b>D</b>). Representative VEP traces from mice prior to OHT (Control), then at 3 d, 14 d, and 28 d after OHT. Data represented combined results from recombinase negative (cre−) and recombinase positive (cre+) CAG-ODD transgenic mice. Days = d, and hours = h.</p> "> Figure 3
<p>Hypoxic neurons in the retina after OHT. Sagittal retinal sections were labeled with neuronal markers (<b>A</b>). RBPMS (RNA binding protein multi-splicing, RGC marker-yellow, arrowhead) showed colocalization (arrowhead) with tdT in GCL. (<b>B</b>). ChAT (anti-choline acetyltransferase, amacrine cell marker, yellow) showed some colocalization with tdT. Arrows are pointing to tdT in Müller glia. Arrowhead is pointing at hypoxic amacrine cells at 6 h, 3 d, and 14 d. (<b>C</b>). PKCα (protein kinase c alpha, bipolar cell marker-yellow, arrowheads) showed some colocalization at 6 h and 3 d. RBPMS (shown in green in this panel). tdT (tdTomato) expression (magenta), DAPI (cell nuclei, blue). Retinal layers are labeled as follows: GCL (ganglion cell layer), INL (inner nuclear layer), and ONL (outer nuclear layer). Scale bar: 25 µm. n = 6 eyes at 6 h, 6 eyes at 3 d, 6 eyes at 14 d, 6 eyes at 28 d, and 10 control eyes. Insets represent a magnification of hypoxic neurons.</p> "> Figure 4
<p>Hypoxic glia in the retina after OHT. Sagittal retinal sections were labeled with glial markers. (<b>A</b>). GFAP (glial fibrillary acidic protein, astrocyte marker, yellow) showed colocalization (arrows) with tdT (magenta) in GCL. (<b>B</b>). Iba-1 (ionized calcium binding adaptor molecule 1, microglia marker, yellow) showed colocalization (arrows) with tdT in INL. (<b>C</b>). Vim vimentin, Müller glia marker, yellow) showed colocalization (arrows) with tdT. (<b>D</b>). Pimonidazole (pimo, hypoxia marker, green, arrows) labeling was reported in Müller glia, and cells in GCL 6 h, 3 d, and 28 d after OHT. Pimonidazole was shown separately since it was overridden by tdT in previous images. tdT (tdTomato) expression (magenta), DAPI (cell nuclei, blue). Retinal layers were labeled as follows: GCL (ganglion cell layer), INL (inner nuclear layer), and ONL (outer nuclear layer). Scale bar: 25 µm. n = 6 eyes at 6 h, 6 eyes at 3 d, 6 eyes at 14 d, 6 eyes at 28 d, and 10 control eyes. Insets represent a magnification of hypoxic glia.</p> "> Figure 5
<p>Hypoxic glia in the ON and ONH after OHT. Sections oriented so the globe is superior. Longitudinal sections of ON showed hypoxic glia detected 6 h, 3 d, 28 d after OHT by tdT expression colocalized with GFAP (yellow) in astrocytes ((<b>A</b>), arrows) and Iba-1 (yellow) in microglia ((<b>B</b>), arrow) compared with control. Pimonidazole-positive glial cells (pimo, green, arrows) were shown colocalized with GFAP in astrocytes (<b>C</b>) and with Iba1 in microglia (<b>D</b>) 6 h and 3 days after OHT. Pimonidazole was shown separately since tdT expression overrode pimonidazole labeling in <b>A</b>,<b>B</b>. ONH showed higher intensity of tdT expression than ON 6hrs and 3d after OHT, which colocalized with GFAP in astrocytes (<b>E</b>) and with Iba-1 in microglia (<b>F</b>). ON: optic nerve, and ONH: optic nerve head. Pimonidazole (pimo, green), tdT = tdT expression (magenta), DAPI (cell nuclei, blue). Scale bar: 25 µm. n = 6 ONHs and ONs at 6 h, 6 ONHs and ONs at 3 d, 6 ONHs and ONs at 14 d, 6 ONHs and ONs at 28 d, and 6 control ONHs and ONs.</p> "> Figure 6
<p>Hypoxia is highest in ONH and ON compared to retina. ON, ONH, and retina at each time point (6 h, 3 d, 14 d, and 28 d) showed significant tdT intensity compared to control (each timepoint and structure vs. baseline (#, <span class="html-italic">p</span> < 0.0001 except the following time points compared to control: 14 d retina (<span class="html-italic">p</span> = 0.0328), 28 d ONH (<span class="html-italic">p</span> = 0.0017), 14 d ONH (<span class="html-italic">p</span> = 0.0030), and 28 d ONH (<span class="html-italic">p</span> = 0.0010). ONH and retina tdT fluorescence intensity is significantly greater at each of the 6 h and 3 d time points as compared to the 14 d and 28 d time points (@<span>$</span>, <span class="html-italic">p</span> = 0.0001). ON tdT fluorescence intensity is significantly greater at 6 h compared to 14 d (@, <span class="html-italic">p</span> < 0.0001) and 28 d (<span>$</span>, <span class="html-italic">p</span> = 0.001) and is also greater at 3 d compared to 14 d (%, <span class="html-italic">p</span> < 0.0001) and 28 d (#, <span class="html-italic">p</span> = 0.0008). n = 6 ONHs and ONs at 6 h, 6 ONHs and ONs at 3 d, 6 ONHs and ONs at 14 d, 6 ONHs and ONs at 28 d, and 6 control ONHs and ONs.</p> "> Figure 7
<p>Hypoxia evident in the visual centers in the brain after OHT. Coronal sections of the brain showed significant hypoxia (tdT: tdTomato expression, magenta, arrows) in the SCN (<b>A</b>), LGN (<b>B</b>), and SC (<b>C</b>) 6 h, and 3 d after OHT compared to control and some td Tomato expression 14 and 28d after OHT. Scale bar: 200 µm. (<b>D</b>). A significant increase (F <sub>(4, 16)</sub> = 28.15) in tdT was detected in glaucomatous SCN 6 h after OHT compared to control (<span>$</span>, <span class="html-italic">p</span> = 0.0005), 14 d (%, <span class="html-italic">p</span> = 0.0007), and 28 d (&, <span class="html-italic">p</span> = 0.0019) after OHT. A significant increase in tdT was shown in glaucomatous SCN 3 d after OHT compared to control, 14 d, and 28 d after OHT (#, <span class="html-italic">p</span> < 0.0001). n = 4 SCNs at 6 h, 5 SCNs at 3 d, 4 SCNs at 14 d, 4 SCNs at 28 d, and 4 control SCNs. (<b>E</b>). A significant increase (F <sub>(4, 16)</sub> = 52.63) in tdT was detected in glaucomatous LGN 6 h and 3 d after OHT compared to control, 14 d, and 28 d after OHT (#, <span class="html-italic">p</span> < 0.0001). n = 4 LGNs at 6 h, 4 LGNs at 3 d, 4 LGNs at 14 d, 5 LGNs at 28 d, and 4 control LGNs. (<b>F</b>). A significant increase (F <sub>(4, 17)</sub> = 14.79) in tdT was shown in glaucomatous SC 6 h after OHT compared to control (%, <span class="html-italic">p</span> =0.0005), 14 d (&, <span class="html-italic">p</span> = 0.0006), and 28 d (!, <span class="html-italic">p</span> = 0.0003) after OHT. A significant increase in tdT was shown in glaucomatous SC 3 d after OHT compared to control (@, <span class="html-italic">p</span> = 0.0033), 14 d (<span>$</span>, <span class="html-italic">p</span> = 0.0044), and 28 d (#, <span class="html-italic">p</span> = 0.0023) after OHT. n = 5 SCs at 6 h, 4 SCs at 3 d, 4 SCs at 14 d, 5 SCs at 28 d, and 4 control SCs. (<b>G</b>). A significant increase (F <sub>(6, 31)</sub> = 27.26) in tdT mean intensity was evident in SCN, LGN, and SC 6 h, 3 d, 14 d, and 28 d after OHT compared with the control (#, <span class="html-italic">p</span> < 0.0001). A significant increase in tdT mean intensity was shown 6h after OHT in LGN compared to 6 h (*, <span class="html-italic">p</span> = 0.0499), and 3 d OHT in the SC (*, <span class="html-italic">p</span> = 0.0143). SCN = suprachiasmatic nucleus, LGN = lateral geniculate nucleus, and SC = superior colliculus.</p> "> Figure 8
<p>Collicular vasculature is hypoxic 6 h after OHT. (<b>A</b>). Immunofluorescence for GFAP (green) in superior colliculus 6h after OHT shows vasculature positive for tdTomato (magenta), indicating ODD stabilization from hypoxia. Cell nuclei labeled with DAPI (blue). 20× magnification; Scale bar = 100 µm. (<b>B</b>). A collicular vessel positive for tdTomato, 60× magnification. Scale bar = 20 µm. (<b>C</b>). A larger vessel in the superior colliculus labeled with tdTomato (magenta), endothelial marker CD-31 (green), and nuclear stain DAPI (blue). 60× magnification. Scale bar = 20 µm. (<b>D</b>). The magenta labeling (alone in grayscale) for the same vessel as (<b>C</b>) with a dotted line (yellow) to show the structure drawn to evaluate colocalization of tdTomato with CD-31; results shown in (<b>E</b>). (<b>E</b>). Colocalization plot for the yellow dotted line in (<b>D</b>); magenta is tdTomato and green is CD-31. Peak correspondence indicated colocalization of the two fluorophores within the vessel. Five brains per group and ten sections per brain were analyzed from the 4 time points (6 h, 3 d, 14 d, 28 d).</p> "> Figure 9
<p>A timecourse of superior colliculus sections after OHT. Superior colliculus was stained using NeuroTrace (green) and DAPI (blue); tdTomato (tdT, magenta) reporter indicates cells subjected to hypoxia. Left panels were imaged at 20× magnification (scale bar = 50 µm) and center panels at 60× magnification (scale bar = 20 µm). Right panels are the tdTomato fluorescence alone, in grayscale. In the control superior colliculus, no tdTomato label (right panel) was observed. tdTomato labeling was most widespread at 6 h and 3 d after OHT, including colocalization with NeuroTrace-positive cells (arrowheads). The dotted line in the 6 h panel is the outline of the 60× center panel. By 14 d, tdTomato labeling was no longer evident in the colliculus. In the 28 d panels, all are at 20× magnification, with the center showing GFAP labeling and the right the tdT. Five brains per group and ten sections per brain were analyzed from the 4 time points (6 h, 3 d, 14 d, 28 d).</p> "> Figure 10
<p>Significant changes in GLUT1 and GLUT3 levels after OHT. (<b>A</b>). Significant GLUT1 level increase in the ON 28 d after OHT compared to 6 h (**, <span class="html-italic">p</span> = 0.0094), and 14 d (**, <span class="html-italic">p</span> = 0.0059). n = 6 ONs at 6 h, 7 ONs at 3 d, 6 ONs at 14 d, 4 ONs at 28 d, and 18 control ONs. (<b>B</b>) Significant GLUT1 level increase in the retina 3 d after OHT compared to control (**, <span class="html-italic">p</span> = 0.0039), 6 h (***, <span class="html-italic">p</span> = 0.0008), and 14 d (*, <span class="html-italic">p</span> = 0.0488). n = 6 retinas at 6 h, 6 retinas at 3 d, 6 retinas at 14 d, 4 retinas at 28 d, and 18 control retinas. (<b>C</b>). Significant GLUT3 level increase in the ON 3 d after OHT compared to 14 d (*, <span class="html-italic">p</span> = 0.0426). n = 5 ONs at 6 h, 7 ONs at 3 d, 6 ONs at 14 d, 5 ONs at 28 d, and 19 control ONs. (<b>D</b>). No significant change in GLUT3 level in the retina 6 h, 3 d, 14 d, and 28 d after OHT (<span class="html-italic">p</span> > 0.05). n = 9 retinas at 6 h, 11 retinas at 3 d, 6 retinas at 14 d, 4 retinas at 28 d, and 18 control retinas.</p> ">
Abstract
:1. Introduction
2. Materials and Methods
2.1. Animals
2.2. Genotyping
2.3. Tamoxifen Injection
2.4. Intraocular Pressure (IOP) Measurements
2.5. Ocular Hypertension (OHT) Model
2.6. Determination of Visual Function
2.7. Hypoxia Detection by Pimonidazole
2.8. Perfusion and Tissue Preparation
2.9. Cryosectioning
2.10. Microtome Sectioning
2.11. Immunofluorescence (IF)
2.12. Histopathology for Light Microscopy
2.13. Quantification of Axons
2.14. Protein Extraction for Capillary-Based Electrophoresis (WES)
2.15. Statistical Analysis
3. Results
3.1. Visually Evoked Potential Significantly Decreased after OHT
3.2. Hypoxic Glia and Neurons in Retina after OHT
3.3. Strong Hypoxia in Optic Nerve (ON) Glia after OHT
3.4. Hypoxia Is Highest in the Optic Nerve Head (ONH) after OHT
3.5. Hypoxia in Brain Visual Nuclei after OHT
3.6. Significant Changes in GLUT1 and GLUT3 Levels after OHT
4. Discussion
4.1. Hypoxic Glia and Neurons in Glaucomatous Retina
4.2. Hypoxic Astrocytes and Microglia in Glaucomatous ON with the Highest Hypoxia in ONH
4.3. Hypoxia in Visual Centers of the Glaucomatous Brain
4.4. The CAG-ODD Reporter System
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Tham, Y.C.; Li, X.; Wong, T.Y.; Quigley, H.A.; Aung, T.; Cheng, C.Y. Global prevalence of glaucoma and projections of glaucoma burden through 2040: A systematic review and meta-analysis. Ophthalmology 2014, 121, 2081–2090. [Google Scholar] [CrossRef] [PubMed]
- Kaur, C.; Sivakumar, V.; Foulds, W. Early Response of Neurons and Glial Cells to Hypoxia in the Retina. Investig. Ophthalmol. Vis. Sci. 2006, 47, 1126–1141. [Google Scholar] [CrossRef] [PubMed]
- Chidlow, G.; Wood, J.P.M.; Casson, R.J. Investigations into Hypoxia and Oxidative Stress at the Optic Nerve Head in a Rat Model of Glaucoma. Front. Neurosci. 2017, 11, 478. [Google Scholar] [CrossRef] [PubMed]
- Koshiji, M.; To, K.K.; Hammer, S.; Kumamoto, K.; Harris, A.L.; Modrich, P.; Huang, L.E. HIF-1alpha induces genetic instability by transcriptionally downregulating MutSalpha expression. Mol. Cell 2005, 17, 793–803. [Google Scholar] [CrossRef]
- Zhang, H.; Bosch-Marce, M.; Shimoda, L.A.; Tan, Y.S.; Baek, J.H.; Wesley, J.B.; Gonzalez, F.J.; Semenza, G.L. Mitochondrial autophagy is an HIF-1-dependent adaptive metabolic response to hypoxia. J. Biol. Chem. 2008, 283, 10892–10903. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Liu, L.; Cash, T.P.; Jones, R.G.; Keith, B.; Thompson, C.B.; Simon, M.C. Hypoxia-induced energy stress regulates mRNA translation and cell growth. Mol. Cell 2006, 21, 521–531. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Jassim, A.H.; Inman, D.M. Evidence of Hypoxic Glial Cells in a Model of Ocular Hypertension. Investig. Ophthalmol. Vis. Sci. 2019, 60, 1–15. [Google Scholar] [CrossRef] [Green Version]
- Kaelin, W.G., Jr.; Ratcliffe, P.J. Oxygen sensing by metazoans: The central role of the HIF hydroxylase pathway. Mol. Cell 2008, 30, 393–402. [Google Scholar] [CrossRef]
- Lin, M.; Chen, Y.; Jin, J.; Hu, Y.; Zhou, K.K.; Zhu, M.; Le, Y.Z.; Ge, J.; Johnson, R.S.; Ma, J.X. Ischaemia-induced retinal neovascularisation and diabetic retinopathy in mice with conditional knockout of hypoxia-inducible factor-1 in retinal Muller cells. Diabetologia 2011, 54, 1554–1566. [Google Scholar] [CrossRef] [Green Version]
- Mowat, F.M.; Luhmann, U.F.; Smith, A.J.; Lange, C.; Duran, Y.; Harten, S.; Shukla, D.; Maxwell, P.H.; Ali, R.R.; Bainbridge, J.W. HIF-1alpha and HIF-2alpha are differentially activated in distinct cell populations in retinal ischaemia. PLoS ONE 2010, 5, e11103. [Google Scholar] [CrossRef]
- Chen, C.; Pore, N.; Behrooz, A.; Ismail-Beigi, F.; Maity, A. Regulation of glut1 mRNA by hypoxia-inducible factor-1. Interaction between H-ras and hypoxia. J. Biol. Chem. 2001, 276, 9519–9525. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Bricker-Anthony, C.; D’Surney, L.; Lunn, B.; Hines-Beard, J.; Jo, M.; Bernardo-Colon, A.; Rex, T.S. Erythropoietin either Prevents or Exacerbates Retinal Damage from Eye Trauma Depending on Treatment Timing. Optom. Vis. Sci. 2017, 94, 20–32. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Ergorul, C.; Ray, A.; Huang, W.; Wang, D.Y.; Ben, Y.; Cantuti-Castelvetri, I.; Grosskreutz, C.L. Hypoxia inducible factor-1alpha (HIF-1alpha) and some HIF-1 target genes are elevated in experimental glaucoma. J. Mol. Neurosci. 2010, 42, 183–191. [Google Scholar] [CrossRef] [Green Version]
- Kimura, W.; Xiao, F.; Canseco, D.C.; Muralidhar, S.; Thet, S.; Zhang, H.M.; Abderrahman, Y.; Chen, R.; Garcia, J.A.; Shelton, J.M.; et al. Hypoxia fate mapping identifies cycling cardiomyocytes in the adult heart. Nature 2015, 523, 226–230. [Google Scholar] [CrossRef] [PubMed]
- Rojas, B.; Gallego, B.I.; Ramírez, A.I.; Salazar, J.J.; de Hoz, R.; Valiente-Soriano, F.; Avilés-Trigueros, M.; Villegas-Perez, M.; Vidal-Sanz, M.; Triviño, A.; et al. Microglia in mouse retina contralateral to experimental glaucoma exhibit multiple signs of activation in all retinal layers. J. Neuroinflamm. 2014, 11, 133. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Coughlin, L.; Morrison, R.S.; Horner, P.J.; Inman, D.M. Mitochondrial morphology differences and mitophagy deficit in murine glaucomatous optic nerve. Investig. Ophthalmol. Vis. Sci. 2015, 56, 1437–1446. [Google Scholar] [CrossRef] [Green Version]
- Kleesattel, D.; Crish, S.D.; Inman, D.M. Decreased Energy Capacity and Increased Autophagic Activity in Optic Nerve Axons With Defective Anterograde Transport. Investig. Ophthalmol. Vis. Sci. 2015, 56, 8215–8227. [Google Scholar] [CrossRef] [Green Version]
- Inman, D.M.; Lambert, W.S.; Calkins, D.J.; Horner, P.J. alpha-Lipoic acid antioxidant treatment limits glaucoma-related retinal ganglion cell death and dysfunction. PLoS ONE 2013, 8, e65389. [Google Scholar] [CrossRef] [Green Version]
- Wilson, G.N.; Smith, M.A.; Inman, D.M.; Dengler-Crish, C.M.; Crish, S.D. Early Cytoskeletal Protein Modifications Precede Overt Structural Degeneration in the DBA/2J Mouse Model of Glaucoma. Front. Neurosci. 2016, 10, 494. [Google Scholar] [CrossRef]
- Coleman, D.J.; Trokel, S. Direct-recorded intraocular pressure variations in a human subject. Arch. Ophthalmol. 1969, 82, 637–640. [Google Scholar] [CrossRef]
- Turner, D.C.; Miranda, M.; Morris, J.S.; Girkin, C.A.; Downs, J.C. Acute Stress Increases Intraocular Pressure in Nonhuman Primates. Ophthalmol. Glaucoma 2019, 2, 210–214. [Google Scholar] [CrossRef] [PubMed]
- Zhi, Z.; Cepurna, W.O.; Johnson, E.C.; Morrison, J.C.; Wang, R.K. Impact of intraocular pressure on changes of blood flow in the retina, choroid, and optic nerve head in rats investigated by optical microangiography. Biomed. Opt. Express 2012, 3, 2220–2233. [Google Scholar] [CrossRef] [PubMed]
- Flammer, J.; Konieczka, K.; Flammer, A.J. The primary vascular dysregulation syndrome: Implications for eye diseases. EPMA J. 2013, 4, 14. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Hannon, B.G.; Feola, A.J.; Gerberich, B.G.; Read, A.T.; Prausnitz, M.R.; Ethier, C.R.; Pardue, M.T. Using retinal function to define ischemic exclusion criteria for animal models of glaucoma. Exp. Eye. Res. 2021, 202, 108354. [Google Scholar] [CrossRef] [PubMed]
- Danhier, P.; Krishnamachary, B.; Bharti, S.; Kakkad, S.; Mironchik, Y.; Bhujwalla, Z.M. Combining Optical Reporter Proteins with Different Half-lives to Detect Temporal Evolution of Hypoxia and Reoxygenation in Tumors. Neoplasia 2015, 17, 871–881. [Google Scholar] [CrossRef] [Green Version]
- Park, H.Y.; Park, J.H.K.a.C.K. Alterations of the synapse of the inner retinal layers after chronic intraocular pressure elevation in glaucoma animal model. Mol. Brain 2014, 7, 53. [Google Scholar] [CrossRef] [Green Version]
- Kanamori, A.; Nakamura, M.; Nakanishi, Y.; Yamada, Y.; Negi, A. Long-term glial reactivity in rat retinas ipsilateral and contralateral to experimental glaucoma. Exp. Eye Res. 2005, 81, 48–56. [Google Scholar] [CrossRef]
- Woldemussie, E.; Wijono, M.; Ruiz, G. Muller cell response to laser-induced increase in intraocular pressure in rats. Glia 2004, 47, 109–119. [Google Scholar] [CrossRef]
- Wu, H.; Chen, Q. Hypoxia activation of mitophagy and its role in disease pathogenesis. Antioxid. Redox Signal. 2015, 22, 1032–1046. [Google Scholar] [CrossRef]
- Papandreou, I.; Cairns, R.A.; Fontana, L.; Lim, A.L.; Denko, N.C. HIF-1 mediates adaptation to hypoxia by actively downregulating mitochondrial oxygen consumption. Cell Metab. 2006, 3, 187–197. [Google Scholar] [CrossRef] [Green Version]
- Kim, J.W.; Tchernyshyov, I.; Semenza, G.L.; Dang, C.V. HIF-1-mediated expression of pyruvate dehydrogenase kinase: A metabolic switch required for cellular adaptation to hypoxia. Cell Metab. 2006, 3, 177–185. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Poitry-Yamate, C.; Poitry, S.; Tsacopoulos, M. Lactate Released by Miiller Glial Cells Is Metabolized by Photoreceptors from Mammalian Retina. J. Neurosci. 1995, 15, 5179–5191. [Google Scholar] [CrossRef] [PubMed]
- Winkler, B.S.; Starnes, C.A.; Sauer, M.W.; Firouzgan, Z.; Chen, S.C. Cultured retinal neuronal cells and Muller cells both show net production of lactate. Neurochem. Int. 2004, 45, 311–320. [Google Scholar] [CrossRef] [PubMed]
- Uga, S.; Smelser, G.K. Comparative study of the fine structure of retinal Müller cells in various vertebrates. Investig. Ophthalmol. 1973, 12, 434–448. [Google Scholar]
- Lindsay, K.J.; Du, J.; Sloat, S.R.; Contreras, L.; Linton, J.D.; Turner, S.J.; Sadilek, M.; Satrustegui, J.; Hurley, J.B. Pyruvate kinase and aspartate-glutamate carrier distributions reveal key metabolic links between neurons and glia in retina. Proc. Natl. Acad. Sci. USA 2014, 111, 15579–15584. [Google Scholar] [CrossRef] [Green Version]
- Eichler, W.; Kuhrt, H.; Hoffmann, S.; Wiedemann, P.; Reichenbach, A. VEGF release by retinal glia depends on both oxygen and glucose supply. Neuroreport 2000, 11, 3533–3537. [Google Scholar] [CrossRef]
- Harun-Or-Rashid, M.; Pappenhagen, N.; Palmer, P.G.; Smith, M.A.; Gevorgyan, V.; Wilson, G.N.; Crish, S.D.; Inman, D.M. Structural and Functional Rescue of Chronic Metabolically Stressed Optic Nerves through Respiration. J. Neurosci. 2018, 38, 5122–5139. [Google Scholar] [CrossRef]
- Sivakumar, V.; Foulds, W.S.; Luu, C.D.; Ling, E.A.; Kaur, C. Retinal ganglion cell death is induced by microglia derived pro-inflammatory cytokines in the hypoxic neonatal retina. J. Pathol. 2011, 224, 245–260. [Google Scholar] [CrossRef]
- Cserep, C.; Posfai, B.; Lenart, N.; Fekete, R.; Laszlo, Z.I.; Lele, Z.; Orsolits, B.; Molnar, G.; Heindl, S.; Schwarcz, A.D.; et al. Microglia monitor and protect neuronal function through specialized somatic purinergic junctions. Science 2020, 367, 528–537. [Google Scholar] [CrossRef]
- Wake, H.; Moorhouse, A.J.; Jinno, S.; Kohsaka, S.; Nabekura, J. Resting microglia directly monitor the functional state of synapses in vivo and determine the fate of ischemic terminals. J. Neurosci. 2009, 29, 3974–3980. [Google Scholar] [CrossRef] [Green Version]
- Kaur, C.; Rathnasamy, G.; Ling, E.A. Roles of activated microglia in hypoxia induced neuroinflammation in the developing brain and the retina. J. Neuroimmune Pharmacol. Off. J. Soc. Neuroimmune Pharmacol. 2013, 8, 66–78. [Google Scholar] [CrossRef] [PubMed]
- Barron, M.J.; Griffiths, P.; Turnbull, D.M.; Bates, D.; Nichols, P. The distributions of mitochondria and sodium channels reflect the specific energy requirements and conduction properties of the human optic nerve head. Br. J. Ophthalmol. 2004, 88, 286–290. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Cwerman-Thibault, H.; Lechauve, C.; Augustin, S.; Roussel, D.; Reboussin, E.; Mohammad, A.; Degardin-Chicaud, J.; Simonutti, M.; Liang, H.; Brignole-Baudouin, F.; et al. Neuroglobin Can Prevent or Reverse Glaucomatous Progression in DBA/2J Mice. Mol. Ther. Methods Clin. Dev. 2017, 5, 200–220. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Wu, J.H.; Zhang, S.H.; Nickerson, J.M.; Gao, F.J.; Sun, Z.; Chen, X.Y.; Zhang, S.J.; Gao, F.; Chen, J.Y.; Luo, Y.; et al. Cumulative mtDNA damage and mutations contribute to the progressive loss of RGCs in a rat model of glaucoma. Neurobiol. Dis. 2015, 74, 167–179. [Google Scholar] [CrossRef] [Green Version]
- Chidlow, G.; Ebneter, A.; Wood, J.P.; Casson, R.J. The optic nerve head is the site of axonal transport disruption, axonal cytoskeleton damage and putative axonal regeneration failure in a rat model of glaucoma. Acta Neuropathol. 2011, 121, 737–751. [Google Scholar] [CrossRef] [Green Version]
- Ghosh, S.; Castillo, E.; Frias, E.S.; Swanson, R.A. Bioenergetic regulation of microglia. Glia 2018, 66, 1200–1212. [Google Scholar] [CrossRef]
- Orihuela, R.; McPherson, C.A.; Harry, G.J. Microglial M1/M2 polarization and metabolic states. Br. J. Pharmacol. 2016, 173, 649–665. [Google Scholar] [CrossRef]
Figure | Experiment | Eye/ON Number |
---|---|---|
1/B | IOP measurement | n = 16 eyes at 3 d, 14 eyes at 14 d, 12 eyes at 28 d, and 30 control eyes |
1/C | RGC count | n = 12 eyes at 6 h, 12 eyes at 3 d, 15 eyes at 14 d, 21 eyes at 28 d, and 20 control eyes. |
1/E | Axon count | n = 7 ONs at 6 h, 6 ONs at 3 d, 8 ONs at 14 d, 7 ONs at 28 d, and 13 control ONs. |
2/A | N1 amplitude | n = 14 eyes at 3 d, 14 eyes at 14 d, 8 eyes at 28 d, and 12 control eyes. |
2/B | P2 amplitude | n = 14 eyes at 3 d, 14 eyes at 14 d, 8 eyes at 28 d, and 12 control eyes |
2/C | Response amplitude | n = 14 eyes at 3 d, 14 eyes at 14 d, 8 eyes at 28 d, and 12 control eyes. |
3 | Retinal hypoxic neurons | n = 6 eyes at 6 h, 6 eyes at 3 d, 6 eyes at 14 d, 6 eyes at 28 d, and 10 control eyes |
4 | Retinal hypoxic glia | n = 6 eyes at 6 h, 6 eyes at 3 d, 6 eyes at 14 d, 6 eyes at 28 d, and 10 control eyes |
5 | Hypoxic glia in ON/ONH | n = 6 ONHs and ONs at 6 h, 6 ONHs and ONs at 3 d, 6 ONHs and ONs at 14 d, 6 ONHs and ONs at 28 d, and 6 control ONHs and ONs |
6 | Highest hypoxia in ONH/ON | n = 6 ONHs and ONs at 6 h, 6 ONHs and ONs at 3 d, 6 ONHs and ONs at 14 d, 6 ONHs and ONs at 28 d, and 6 control ONHs and ONs |
7/A–D | Hypoxia in SCN | n = 4 SCNs at 6 h, 5 SCNs at 3 d, 4 SCNs at 14 d, 4 SCNs at 28 d, and 4 control SCNs. |
7/E | Hypoxia in LGN | n = 4 LGNs at 6 h, 4 LGNs at 3 d, 4 LGNs at 14 d, 5 LGNs at 28 d, and 4 control LGNs |
7/F | Hypoxia in SC | n = 5 SCs at 6 h, 4 SCs at 3 d, 4 SCs at 14 d, 5 SCs at 28 d, and 4 control SCs |
8 | Hypoxia in SC | n = 5 SCs at 6 h, 3 d, 14 d, 28 d; 10 sections per SC |
9 | Hypoxia in SC | n = 5 SCs at 6 h, 3 d, 14 d, 28 d; 10 sections per SC |
10/A | GLUT1 increase in ON | n = 6 ONs at 6 h, 7 ONs at 3 d, 6 ONs at 14 d, 4 ONs at 28 d, and 18 control ONs |
10/B | GLUT1 increase in retina | n = 6 retinas at 6 h, 6 retinas at 3 d, 6 retinas at 14 d, 4 retinas at 28 d, and 18 control retinas |
10/C | GLUT3 increase in ON | n = 5 ONs at 6 h, 7 ONs at 3 d, 6 ONs at 14 d, 5 ONs at 28 d, and 19 control ONs |
10/D | No change of GLUT3 in retina | n = 9 retinas at 6 h, 11 retinas at 3 d, 6 retinas at 14 d, 4 retinas at 28 d, and 18 control retinas |
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Jassim, A.H.; Nsiah, N.Y.; Inman, D.M. Ocular Hypertension Results in Hypoxia within Glia and Neurons throughout the Visual Projection. Antioxidants 2022, 11, 888. https://doi.org/10.3390/antiox11050888
Jassim AH, Nsiah NY, Inman DM. Ocular Hypertension Results in Hypoxia within Glia and Neurons throughout the Visual Projection. Antioxidants. 2022; 11(5):888. https://doi.org/10.3390/antiox11050888
Chicago/Turabian StyleJassim, Assraa Hassan, Nana Yaa Nsiah, and Denise M. Inman. 2022. "Ocular Hypertension Results in Hypoxia within Glia and Neurons throughout the Visual Projection" Antioxidants 11, no. 5: 888. https://doi.org/10.3390/antiox11050888