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Cell and Tissue Research

, Volume 336, Issue 1, pp 21–29 | Cite as

Developmental profile of erythropoietin and its receptor in guinea-pig retina

  • Kathryn Munro
  • Sandra Rees
  • Rachael O’Dowd
  • Mary Tolcos
Regular Article

Abstract

Evidence suggests that endogenous erythropoietin (EPO) is involved in the development of the central nervous system; however, its role in retinal development is yet to be determined. In this study, we have used fluorescence immunohistochemistry to localise EPO and its receptor (EPOR) in the developing and mature retina of the guinea-pig, a species in which retinal development is similar to that in humans. EPO immunoreactivity (IR) was observed in ganglion cells from 25 days of gestation (dg; term ∼67 dg), and in the inner and outer plexiform layers and in horizontal cells by 40 dg. EPO-IR persisted in all of these structures into adulthood. Müller cells also displayed EPO-IR, which was seen in the radial processes and endfeet at 40 dg and in the cytoplasm by 50 dg. IR in these cells was particularly intense and appeared to increase with age. EPOR-IR was found in all ages examined; it was detected in ganglion cells at 25 dg and, from 30 dg onwards, was localised on, and adjacent to, the cell surface membrane. The distribution of EPOR-IR became increasingly widespread during gestation and, by 50 dg, EPOR-IR was detectable on the majority of retinal somal membranes. This localisation persisted in the postnatal and adult retina. Therefore, IR for EPO and its receptor is present in the guinea-pig retina from as early as 25 dg, when retinal layers are forming, and persists throughout postnatal development. This suggests that EPO plays a role both in retinal development and in the maintenance of the adult retina.

Keywords

Erythropoietin Erythropoietin receptor Retina Ontogeny Guinea-pig 

Abbreviations

CNS

central nervous system

dg

days of gestation

EPO

erythropoietin

EPOR

erythropoietin receptor

GCL

ganglion cell layer

GS

glutamine synthetase

ILM

inner limiting membrane

INL

inner nuclear layer

IPL

inner plexiform layer

IR

immunoreactivity

NFL

nerve fibre layer

OLM

outer limiting membrane

ONL

outer nuclear layer

OPL

outer plexiform layer

P

postnatal day

PB

phosphate buffer

PBS

phosphate-buffered saline

PRL

photoreceptor layer

TBS-T

TRIS-buffered saline with Triton X-100

Notes

Acknowledgements

The authors thank Dr. Mark Ransome for his assistance with the EPOR immunoprecipitation assay.

References

  1. Bernaudin M, Marti HH, Roussel S, Divoux D, Nouvelot A, MacKenzie ET, Petit E (1999) A potential role for erythropoietin in focal permanent cerebral ischemia in mice. J Cereb Blood Flow Metab 19:643–651PubMedCrossRefGoogle Scholar
  2. Bernaudin M, Bellail A, Marti HH, Yvon A, Vivien D, Duchatelle I, Mackenzie ET, Petit E (2000) Neurons and astrocytes express EPO mRNA: oxygen-sensing mechanisms that involve the redox-state of the brain. Glia 30:271–278PubMedCrossRefGoogle Scholar
  3. Bocker-Meffert S, Rosenstiel P, Rohl C, Warneke N, Held-Feindt J, Sievers J, Lucius R (2002) Erythropoietin and VEGF promote neural outgrowth from retinal explants in postnatal rats. Invest Ophthalmol Vis Sci 43:2021–2026PubMedGoogle Scholar
  4. Brines ML, Ghezzi P, Keenan S, Agnello D, de Lanerolle NC, Cerami C, Itri LM, Cerami A (2000) Erythropoietin crosses the blood-brain barrier to protect against experimental brain injury. Proc Natl Acad Sci USA 97:10526–10531PubMedCrossRefGoogle Scholar
  5. Caye-Thomasen P, Wagner N, Lidegaard Frederiksen B, Asal K, Thomsen J (2005) Erythropoietin and erythropoietin receptor expression in the guinea pig inner ear. Hear Res 203:21–27PubMedCrossRefGoogle Scholar
  6. Digicaylioglu M, Bichet S, Marti HH, Wenger RH, Rivas LA, Bauer C, Gassmann M (1995) Localization of specific erythropoietin binding sites in defined areas of the mouse brain. Proc Natl Acad Sci USA 92:3717–3720PubMedCrossRefGoogle Scholar
  7. Eckardt KU, Kurtz A (2005) Regulation of erythropoietin production. Eur J Clin Invest 35 (Suppl 3):13–19PubMedCrossRefGoogle Scholar
  8. Garcia-Ramirez M, Hernandez C, Simo R (2008) Expression of erythropoietin and its receptor in the human retina: a comparative study of diabetic and nondiabetic subjects. Diabetes Care 31:1189–1194PubMedCrossRefGoogle Scholar
  9. Grimm C, Wenzel A, Groszer M, Mayser H, Seeliger M, Samardzija M, Bauer C, Gassmann M, Reme CE (2002) HIF-1-induced erythropoietin in the hypoxic retina protects against light-induced retinal degeneration. Nat Med 8:718–724PubMedCrossRefGoogle Scholar
  10. Grimm C, Wenzel A, Stanescu D, Samardzija M, Hotop S, Groszer M, Naash M, Gassmann M, Reme C (2004) Constitutive overexpression of human erythropoietin protects the mouse retina against induced but not inherited retinal degeneration. J Neurosci 24:5651–5658PubMedCrossRefGoogle Scholar
  11. Junk AK, Mammis A, Savitz SI, Singh M, Roth S, Malhotra S, Rosenbaum PS, Cerami A, Brines M, Rosenbaum DM (2002) Erythropoietin administration protects retinal neurons from acute ischemia-reperfusion injury. Proc Natl Acad Sci USA 99:10659–10664PubMedCrossRefGoogle Scholar
  12. Juul SE, Anderson DK, Li Y, Christensen RD (1998) Erythropoietin and erythropoietin receptor in the developing human central nervous system. Pediatr Res 43:40–49PubMedCrossRefGoogle Scholar
  13. Knabe W, Knerlich F, Washausen S, Kietzmann T, Siren AL, Brunnett G, Kuhn HJ, Ehrenreich H (2004) Expression patterns of erythropoietin and its receptor in the developing midbrain. Anat Embryol (Berl) 207:503–512CrossRefGoogle Scholar
  14. Loeliger M, Briscoe T, Lambert G, Caddy J, Rehn A, Dieni S, Rees S (2004) Chronic placental insufficiency affects retinal development in the guinea pig. Invest Ophthalmol Vis Sci 45:2361–2367PubMedCrossRefGoogle Scholar
  15. Loeliger M, Duncan J, Louey S, Cock M, Harding R, Rees S (2005) Fetal growth restriction induced by chronic placental insufficiency has long-term effects on the retina but not the optic nerve. Invest Ophthalmol Vis Sci 46:3300–3308PubMedCrossRefGoogle Scholar
  16. Marti HH, Wenger RH, Rivas LA, Straumann U, Digicaylioglu M, Henn V, Yonekawa Y, Bauer C, Gassmann M (1996) Erythropoietin gene expression in human, monkey and murine brain. Eur J Neurosci 8:666–676PubMedCrossRefGoogle Scholar
  17. Masuda S, Okano M, Yamagishi K, Nagao M, Ueda M, Sasaki R (1994) A novel site of erythropoietin production. Oxygen-dependent production in cultured rat astrocytes. J Biol Chem 269:19488–19493PubMedGoogle Scholar
  18. Morishita E, Masuda S, Nagao M, Yasuda Y, Sasaki R (1997) Erythropoietin receptor is expressed in rat hippocampal and cerebral cortical neurons, and erythropoietin prevents in vitro glutamate-induced neuronal death. Neuroscience 76:105–116PubMedCrossRefGoogle Scholar
  19. Patel S, Rowe MJ, Winters SA, Ohls RK (2008) Elevated erythropoietin mRNA and protein concentrations in the developing human eye. Pediatr Res 63:394–397PubMedCrossRefGoogle Scholar
  20. Pillai A, Mahadik SP (2006) Differential effects of haloperidol and olanzapine on the expression of erythropoietin and its receptor in rat hippocampus and striatum. J Neurochem 98:1411–1422PubMedCrossRefGoogle Scholar
  21. Rees S, Bainbridge A (1992) The structural and neurochemical development of the fetal guinea pig retina and optic nerve in experimental growth retardation. Int J Dev Neurosci 10:93–108PubMedCrossRefGoogle Scholar
  22. Schnitzer J (1988) Astrocytes in the guinea pig, horse, and monkey retina: their occurrence coincides with the presence of blood vessels. Glia 1:74–89PubMedCrossRefGoogle Scholar
  23. Shingo T, Sorokan ST, Shimazaki T, Weiss S (2001) Erythropoietin regulates the in vitro and in vivo production of neuronal progenitors by mammalian forebrain neural stem cells. J Neurosci 21:9733–9743PubMedGoogle Scholar
  24. Siren AL, Knerlich F, Poser W, Gleiter CH, Bruck W, Ehrenreich H (2001) Erythropoietin and erythropoietin receptor in human ischemic/hypoxic brain. Acta Neuropathol 101:271–276PubMedGoogle Scholar
  25. Spira AW (1975) In utero development and maturation of the retina of a non-primate mammal: a light and electron microscopic study of the guinea pig. Anat Embryol (Berl) 146:279–300CrossRefGoogle Scholar
  26. Tsai PT, Ohab JJ, Kertesz N, Groszer M, Matter C, Gao J, Liu X, Wu H, Carmichael ST (2006) A critical role of erythropoietin receptor in neurogenesis and post-stroke recovery. J Neurosci 26:1269–1274PubMedCrossRefGoogle Scholar
  27. Weishaupt JH, Rohde G, Polking E, Siren AL, Ehrenreich H, Bahr M (2004) Effect of erythropoietin axotomy-induced apoptosis in rat retinal ganglion cells. Invest Ophthalmol Vis Sci 45:1514–1522PubMedCrossRefGoogle Scholar
  28. Yamaji R, Okada T, Moriya M, Naito M, Tsuruo T, Miyatake K, Nakano Y (1996) Brain capillary endothelial cells express two forms of erythropoietin receptor mRNA. Eur J Biochem 239:494–500PubMedCrossRefGoogle Scholar
  29. Yamasaki M, Mishima HK, Yamashita H, Kashiwagi K, Murata K, Minamoto A, Inaba T (2005) Neuroprotective effects of erythropoietin on glutamate and nitric oxide toxicity in primary cultured retinal ganglion cells. Brain Res 1050:15–26PubMedCrossRefGoogle Scholar
  30. Yasuda Y, Nagao M, Okano M, Masuda S, Sasaki R, Konishi H, Tamnimura T (1993) Localization of erythropoietin and erythropoietin-receptor in postimplantation mouse embryos. Dev Growth Differ 35:711–722CrossRefGoogle Scholar
  31. Yu DY, Cringle SJ (2001) Oxygen distribution and consumption within the retina in vascularised and avascular retinas and in animal models of retinal disease. Prog Retin Eye Res 20:175–208PubMedCrossRefGoogle Scholar
  32. Yu DY, Cringle SJ, Alder VA, Su EN, Yu PK (1996) Intraretinal oxygen distribution and choroidal regulation in the avascular retina of guinea pigs. Am J Physiol 270:H965–H973PubMedGoogle Scholar
  33. Yu X, Shacka JJ, Eells JB, Suarez-Quian C, Przygodzki RM, Beleslin-Cokic B, Lin CS, Nikodem VM, Hempstead B, Flanders KC, Costantini F, Noguchi CT (2002) Erythropoietin receptor signalling is required for normal brain development. Development 129:505–516PubMedGoogle Scholar
  34. Zhu B, Wang W, Gu Q, Xu X (2008) Erythropoietin protects retinal neurons and glial cells in early-stage streptozotocin-induced diabetic rats. Exp Eye Res 86:375–382PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Kathryn Munro
    • 1
  • Sandra Rees
    • 1
  • Rachael O’Dowd
    • 1
  • Mary Tolcos
    • 1
    • 2
  1. 1.Department of Anatomy and Cell BiologyThe University of MelbourneMelbourneAustralia
  2. 2.Department of Anatomy and Cell BiologyThe University of MelbourneParkvilleAustralia

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