Abstract
Low levels of hypoxia have been suggested to be a mechanism of retinal damage in glaucoma. To test the hypothesis that the activation of the hypoxia-responsive transcription factor hypoxia inducible factor-1α (HIF-1α) is involved in the pathophysiology of glaucoma, we used a rat model of glaucoma to study (1) HIF-1α retinal protein levels by immunoblot analysis, (2) cellular localization of HIF-1α in the retina by immunohistochemistry, and (3) expression of retinal HIF-1 gene targets by quantitative real-time polymerase chain reaction. Glaucoma was unilaterally induced in rats by injecting hypertonic saline in episcleral veins. We find that HIF-1α protein was increased in the retina following elevation of intraocular pressure, specifically in Müller glia and astrocytes but not in activated microglia. Eight established HIF-1 target genes were measured in experimental glaucoma. Retinal Epo, Flt-1, Hsp-27, Pai-1, and Vegfa mRNA levels were increased and Et-1, Igf2, and Tgfβ3 levels were decreased in the glaucomatous retinas. Thus, the increase in HIF-1α levels in Müller glia and astrocytes is accompanied by a marked up regulation of some, but not all, HIF-1 transcriptional targets. These data support the hypothesis that HIF-1α becomes transcriptionally active in astrocytes and Müller cells but not microglia or neurons in glaucoma, arguing against a global hypoxia stimulus to the retina.
Similar content being viewed by others
References
Ahmed F, Brown KM, Stephan DA, Morrison JC, Johnson EC, Tomarev SI (2004) Microarray analysis of changes in mRNA levels in the rat retina after experimental elevation of intraocular pressure. Invest Ophthalmol Vis Sci 45(4):1247–1258
Benn SC, Perrelet D, Kato AC et al (2002) Hsp27 upregulation and phosphorylation is required for injured sensory and motor neuron survival. Neuron 36(1):45–56
Bilton RL, Booker GW (2003) The subtle side to hypoxia inducible factor (HIFalpha) regulation. Eur J Biochem 270(5):791–798
Brahimi-Horn MC, Pouyssegur J (2009) HIF at a glance. J Cell Sci 122(Pt 8):1055–1057
Caniggia I, Mostachfi H, Winter J et al (2000) Hypoxia-inducible factor-1 mediates the biological effects of oxygen on human trophoblast differentiation through TGFbeta(3). J Clin Invest 105(5):577–587
Damert A, Ikeda E, Risau W (1997) Activator-protein-1 binding potentiates the hypoxia-induciblefactor-1-mediated hypoxia-induced transcriptional activation of vascular-endothelial growth factor expression in C6 glioma cells. Biochem J 327(Pt 2):419–423
Dan J, Belyea D, Gertner G, Leshem I, Lusky M, Miskin R (2005) Plasminogen activator inhibitor-1 in the aqueous humor of patients with and without glaucoma. Arch Ophthalmol 123(2):220–224
Dery MA, Michaud MD, Richard DE (2005) Hypoxia-inducible factor 1: regulation by hypoxic and non-hypoxic activators. Int J Biochem Cell Biol 37(3):535–540
Emre M, Orgul S, Haufschild T, Shaw SG, Flammer J (2005) Increased plasma endothelin-1 levels in patients with progressive open angle glaucoma. Br J Ophthalmol 89(1):60–63
Ergorul C, Ray A, Huang W, Darland D, Luo ZK, Grosskreutz CL (2008) Levels of vascular endothelial growth factor-A165b (VEGF-A165b) are elevated in experimental glaucoma. Mol Vis 14:1517–1524
Farkas RH, Chowers I, Hackam AS et al (2004) Increased expression of iron-regulating genes in monkey and human glaucoma. Invest Ophthalmol Vis Sci 45(5):1410–1417
Feke GT, Pasquale LR (2008) Retinal blood flow response to posture change in glaucoma patients compared with healthy subjects. Ophthalmology 115(2):246–252
Feldser D, Agani F, Iyer NV, Pak B, Ferreira G, Semenza GL (1999) Reciprocal positive regulation of hypoxia-inducible factor 1alpha and insulin-like growth factor 2. Cancer Res 59(16):3915–3918
Fu QL, Wu W, Wang H, Li X, Lee VW, So KF (2008) Up-regulated endogenous erythropoietin/erythropoietin receptor system and exogenous erythropoietin rescue retinal ganglion cells after chronic ocular hypertension. Cell Mol Neurobiol 28(2):317–329
Gerber HP, Condorelli F, Park J, Ferrara N (1997) Differential transcriptional regulation of the two vascular endothelial growth factor receptor genes. Flt-1, but not Flk-1/KDR, is up-regulated by hypoxia. J Biol Chem 272(38):23659–23667
Glass CA, Harper SJ, Bates DO (2006) The anti-angiogenic VEGF isoform VEGF165b transiently increases hydraulic conductivity, probably through VEGF receptor 1 in vivo. J Physiol 572(Pt 1):243–257
Grimm C, Hermann DM, Bogdanova A et al (2005) Neuroprotection by hypoxic preconditioning: HIF-1 and erythropoietin protect from retinal degeneration. Semin Cell Dev Biol 16(4–5):531–538
Harris A, Rechtman E, Siesky B, Jonescu-Cuypers C, McCranor L, Garzozi HJ (2005) The role of optic nerve blood flow in the pathogenesis of glaucoma. Ophthalmol Clin North Am 18(3):345–353
Hu J, Discher DJ, Bishopric NH, Webster KA (1998) Hypoxia regulates expression of the endothelin-1 gene through a proximal hypoxia-inducible factor-1 binding site on the antisense strand. Biochem Biophys Res Commun 245(3):894–899
Hu DN, Ritch R, Liebmann J, Liu Y, Cheng B, Hu MS (2002) Vascular endothelial growth factor is increased in aqueous humor of glaucomatous eyes. J Glaucoma 11(5):406–410
Huang W, Fileta JB, Filippopoulos T, Ray A, Dobberfuhl A, Grosskreutz CL (2007) Hsp27 phosphorylation in experimental glaucoma. Invest Ophthalmol Vis Sci 48(9):4129–4135
Kaiser HJ, Flammer J, Wenk M, Luscher T (1995) Endothelin-1 plasma levels in normal-tension glaucoma: abnormal response to postural changes. Graefes Arch Clin Exp Ophthalmol 233(8):484–488
Kalesnykas G, Niittykoski M, Rantala J et al (2007) The expression of heat shock protein 27 in retinal ganglion and glial cells in a rat glaucoma model. Neuroscience 150(3):692–704
Kallberg ME, Brooks DE, Gelatt KN, Garcia-Sanchez GA, Szabo NJ, Lambrou GN (2007) Endothelin-1, nitric oxide, and glutamate in the normal and glaucomatous dog eye. Vet Ophthalmol 10(Suppl 1):46–52
Ke Q, Costa M (2006) Hypoxia-inducible factor-1 (HIF-1). Mol Pharmacol 70(5):1469–1480
Kietzmann T, Samoylenko A, Roth U, Jungermann K (2003) Hypoxia-inducible factor-1 and hypoxia response elements mediate the induction of plasminogen activator inhibitor-1 gene expression by insulin in primary rat hepatocytes. Blood 101(3):907–914
Kumada M, Niwa M, Hara A et al (2005) Tissue type plasminogen activator facilitates NMDA-receptor-mediated retinal apoptosis through an independent fibrinolytic cascade. Invest Ophthalmol Vis Sci 46(4):1504–1507
Latchman DS (2005) HSP27 and cell survival in neurones. Int J Hyperthermia 21(5):393–402
Lupien C, Brenner M, Guerin SL, Salesse C (2004) Expression of glial fibrillary acidic protein in primary cultures of human Muller cells. Exp Eye Res 79(3):423–429
Mali RS, Cheng M, Chintala SK (2005) Plasminogen activators promote excitotoxicity-induced retinal damage. FASEB J 19(10):1280–1289
Mazurek B, Rheinlander C, Fuchs FU et al (2006) Influence of ischemia/hypoxia on the HIF-1 activity and expression of hypoxia-dependent genes in the cochlea of the newborn rat. HNO 54(9):689–697
Meijerink J, Mandigers C, van de Locht L, Tonnissen E, Goodsaid F, Raemaekers J (2001) A novel method to compensate for different amplification efficiencies between patient DNA samples in quantitative real-time PCR. J Mol Diagn 3(2):55–61
Miyahara T, Kikuchi T, Akimoto M, Kurokawa T, Shibuki H, Yoshimura N (2003) Gene microarray analysis of experimental glaucomatous retina from cynomologous monkey. Invest Ophthalmol Vis Sci 44(10):4347–4356
Moore CG, Milne ST, Morrison JC (1993) Noninvasive measurement of rat intraocular pressure with the Tono-Pen. Invest Ophthalmol Vis Sci 34(2):363–369
Morrison JC, Moore CG, Deppmeier LM, Gold BG, Meshul CK, Johnson EC (1997) A rat model of chronic pressure-induced optic nerve damage. Exp Eye Res 64(1):85–96
Nicolela MT (2008) Clinical clues of vascular dysregulation and its association with glaucoma. Can J Ophthalmol 43(3):337–341
Nicolela MT, Ferrier SN, Morrison CA et al (2003) Effects of cold-induced vasospasm in glaucoma: the role of endothelin-1. Invest Ophthalmol Vis Sci 44(6):2565–2572
Peyssonnaux C, Nizet V, Johnson RS (2008) Role of the hypoxia inducible factors HIF in iron metabolism. Cell Cycle 7(1):28–32
Prasanna G, Hulet C, Desai D et al (2005) Effect of elevated intraocular pressure on endothelin-1 in a rat model of glaucoma. Pharmacol Res 51(1):41–50
Richard DE, Berra E, Gothie E, Roux D, Pouyssegur J (1999) p42/p44 mitogen-activated protein kinases phosphorylate hypoxia-inducible factor 1alpha (HIF-1alpha) and enhance the transcriptional activity of HIF-1. J Biol Chem 274(46):32631–32637
Savagian CA, Dubielzig RR, Nork TM (2008) Comparison of the distribution of glial fibrillary acidic protein, heat shock protein 60, and hypoxia-inducible factor-1alpha in retinas from glaucomatous and normal canine eyes. Am J Vet Res 69(2):265–272
Semenza GL (2000) HIF-1: mediator of physiological and pathophysiological responses to hypoxia. J Appl Physiol 88(4):1474–1480
Semenza GL (2001) Hypoxia-inducible factor 1: oxygen homeostasis and disease pathophysiology. Trends Mol Med 7(8):345–350
Stasi K, Nagel D, Yang X, Ren L, Mittag T, Danias J (2007) Ceruloplasmin upregulation in retina of murine and human glaucomatous eyes. Invest Ophthalmol Vis Sci 48(2):727–732
Stockmann C, Fandrey J (2006) Hypoxia-induced erythropoietin production: a paradigm for oxygen-regulated gene expression. Clin Exp Pharmacol Physiol 33(10):968–979
Tezel G, Wax MB (2004) Hypoxia-inducible factor 1alpha in the glaucomatous retina and optic nerve head. Arch Ophthalmol 122(9):1348–1356
Tezel G, Kass MA, Kolker AE, Becker B, Wax MB (1997) Plasma and aqueous humor endothelin levels in primary open-angle glaucoma. J Glaucoma 6(2):83–89
Tsai FJ, Lin HJ, Chen WC, Chen HY, Fan SS (2003) Insulin-like growth factor-II gene polymorphism is associated with primary open angle glaucoma. J Clin Lab Anal 17(6):259–263
Wakakura M, Foulds WS (1989) Response of cultured Muller cells to heat shock—an immunocytochemical study of heat shock and intermediate filament proteins in response to temperature elevation. Exp Eye Res 48(3):337–350
Whitlock NA, Agarwal N, Ma JX, Crosson CE (2005) Hsp27 upregulation by HIF-1 signaling offers protection against retinal ischemia in rats. Invest Ophthalmol Vis Sci 46(3):1092–1098
Woolard J, Wang WY, Bevan HS et al (2004) VEGF165b, an inhibitory vascular endothelial growth factor splice variant: mechanism of action, in vivo effect on angiogenesis and endogenous protein expression. Cancer Res 64(21):7822–7835
Yee Koh M, Spivak-Kroizman TR, Powis G (2008) HIF-1 regulation: not so easy come, easy go. Trends Biochem Sci 33(11):526–534
Yoneda K, Nakano M, Mori K, Kinoshita S, Tashiro K (2007) Disease-related quantitation of TGF-beta3 in human aqueous humor. Growth Factors 25(3):160–167
Acknowledgments
The authors thank Hisatomo Kowa and Bradley Hyman at MassGeneral Institute for Neurodegeneration, Charlestown, MA for the discussions and for the equipment used in qRT-PCR. The authors also thank Charles Vanderburg, Rachel Diamond, and Ozge Cagsal-Getkin at the Advanced Tissue Resource Center/MGH, Charlestown, MA for their technical help. This work was supported by R01-EY13399 (CLG), T32-EY07145 (CE), Massachusetts Lions Grant (CLG), and MEEI Vision-Core Grant EY014104.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Ergorul, C., Ray, A., Huang, W. et al. Hypoxia Inducible Factor-1α (HIF-1α) and Some HIF-1 Target Genes are Elevated in Experimental Glaucoma. J Mol Neurosci 42, 183–191 (2010). https://doi.org/10.1007/s12031-010-9343-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12031-010-9343-z