Abstract
The intravitreal injection of N-methyl-d-aspartate (NMDA), a glutamate analogue, is an established model for fast retinal ganglion cell (RGC) degeneration. Yet, NMDA does not cause specific RGC damage. Now, the effects on the whole retina were analyzed. Additionally, the related effects for the structure and apoptotic levels of the optic nerve were investigated. Therefore, different NMDA concentrations were intravitreally injected in rats (20, 40, or 80 nmol NMDA or PBS). At days 3 and 14, Brn-3a+ RGCs were degenerated. A damage of calretinin+ amacrine cells was also recognized at day 14. Only a slight damage was observed in regard to PKCα+ bipolar cells, while rhodopsin+ photoreceptors remained intact. A long-lasting retinal microglia response was observed from day 3 up to day 14. Furthermore, a partial degeneration of the optic nerve was noted. At day 3, the SMI-32+ neurofilaments were just slightly affected, whereas the neurofilament structure was further degenerated at day 14. However, the luxol fast blue (LFB)-stained myelin structure remained intact from day 3 up to day 14. Interestingly, apoptotic mechanisms, like FasL and Fas co-localization as well as caspase 3 activation, were restricted to the optic nerve of the highest NMDA group at this late stage of degeneration. The degeneration of the optic nerve is probably only a side effect of neuronal degeneration of the inner retinal layers. The intact myelin structure might form a barrier against the direct influence of NMDA. In conclusion, this model is very suitable to test therapeutic agents, but it is important to analyze all inner retina layers and the optic nerve to determine their efficacy in this model more precisely.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- AIF:
-
Apoptosis-inducing factor
- BCA:
-
Bicinchoninic acid assay
- FasL:
-
Fas ligand
- Fas:
-
Fas receptor
- GFAP:
-
Glial fibrillary acid protein
- HE:
-
Hematoxylin and eosin
- IOP:
-
Intraocular pressure
- LIF:
-
Leukemia inhibitory factor
- MAPK:
-
MAP kinase
- NMDA:
-
N-Methyl-d-aspartate
- NMDA-R:
-
N-Methyl-d-aspartate receptor
- PKCα:
-
Protein kinase Cα
- RGCs:
-
Retinal ganglion cells
References
Agca C et al (2013) p38 MAPK signaling acts upstream of LIF-dependent neuroprotection during photoreceptor degeneration. Cell Death Dis 4:e785. https://doi.org/10.1038/cddis.2013.323
Akopian A, Kumar S, Ramakrishnan H, Viswanathan S, Bloomfield SA (2016) Amacrine cells coupled to ganglion cells via gap junctions are highly vulnerable in glaucomatous mouse retinas. J Comp Neurol. https://doi.org/10.1002/cne.24074
Alix JJ, Domingues AM (2011) White matter synapses: form, function, and dysfunction. Neurology 76:397–404. https://doi.org/10.1212/WNL.0b013e3182088273
Biermann J, van Oterendorp C, Stoykow C, Volz C, Jehle T, Boehringer D, Lagreze WA (2012) Evaluation of intraocular pressure elevation in a modified laser-induced glaucoma rat model. Exp Eye Res 104:7–14. https://doi.org/10.1016/j.exer.2012.08.011
Bosco A, Romero CO, Breen KT, Chagovetz AA, Steele MR, Ambati BK, Vetter ML (2015) Neurodegeneration severity is anticipated by early microglia alterations monitored in vivo in a mouse model of chronic glaucoma. Dis Model Mech. https://doi.org/10.1242/dmm.018788
Casola C, Reinehr S, Kuehn S, Stute G, Spiess BM, Dick HB, Joachim SC (2016) Specific inner retinal layer cell damage in an autoimmune glaucoma model is induced by GDNF with or without HSP27. Invest Ophthalmol Vis Sci 57:3626–3639. https://doi.org/10.1167/iovs.15-18999R2
Chidlow G, Ebneter A, Wood JP, Casson RJ (2011) 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 121:737–751. https://doi.org/10.1007/s00401-011-0807-1
Chintala S, Cheng M, Zhang X (2015) Decreased expression of DREAM promotes the degeneration of retinal neurons. PLoS One 10:e0127776. https://doi.org/10.1371/journal.pone.0127776
Choi C, Benveniste EN (2004) Fas ligand/Fas system in the brain: regulator of immune and apoptotic responses. Brain Res Brain Res Rev 44:65–81
Cuenca N et al (2010) Changes in the inner and outer retinal layers after acute increase of the intraocular pressure in adult albino Swiss mice. Exp Eye Res 91:273–285. https://doi.org/10.1016/j.exer.2010.05.020
Di Pierdomenico J, Garcia-Ayuso D, Pinilla I, Cuenca N, Vidal-Sanz M, Agudo-Barriuso M, Villegas-Perez MP (2017) Early events in retinal degeneration caused by rhodopsin mutation or pigment epithelium malfunction: differences and similarities. Front Neuroanat 11:14. https://doi.org/10.3389/fnana.2017.00014
Dong XX, Wang Y, Qin ZH (2009) Molecular mechanisms of excitotoxicity and their relevance to pathogenesis of neurodegenerative diseases. Acta Pharmacol Sin 30:379–387. https://doi.org/10.1038/aps.2009.24
Dreyer EB, Pan ZH, Storm S, Lipton SA (1994) Greater sensitivity of larger retinal ganglion cells to NMDA-mediated cell death. Neuroreport 5:629–631
Dreyer EB, Zurakowski D, Schumer RA, Podos SM, Lipton SA (1996) Elevated glutamate levels in the vitreous body of humans and monkeys with glaucoma. Arch Ophthalmol 114:299–305
Duan Y, Gross RA, Sheu SS (2007) Ca2+-dependent generation of mitochondrial reactive oxygen species serves as a signal for poly(ADP-ribose) polymerase-1 activation during glutamate excitotoxicity. J Physiol 585:741–758. https://doi.org/10.1113/jphysiol.2007.145409
Esquiva G, Avivi A, Hannibal J (2016) Non-image forming light detection by melanopsin, rhodopsin, and long-middlewave (L/W) cone opsin in the subterranean blind mole rat, Spalax ehrenbergi: immunohistochemical characterization, distribution, and connectivity. Front Neuroanat 10:61. https://doi.org/10.3389/fnana.2016.00061
Greferath U, Grunert U, Wassle H (1990) Rod bipolar cells in the mammalian retina show protein kinase C-like immunoreactivity. J Comp Neurol 301:433–442. https://doi.org/10.1002/cne.903010308
Gregory MS et al (2011) Opposing roles for membrane bound and soluble Fas ligand in glaucoma-associated retinal ganglion cell death. PLoS One 6:e17659. https://doi.org/10.1371/journal.pone.0017659
Horstmann L, Schmid H, Heinen AP, Kurschus FC, Dick HB, Joachim SC (2013) Inflammatory demyelination induces glia alterations and ganglion cell loss in the retina of an experimental autoimmune encephalomyelitis model. J Neuroinflammation 10:120. https://doi.org/10.1186/1742-2094-10-120
Ito Y, Shimazawa M, Inokuchi Y, Fukumitsu H, Furukawa S, Araie M, Hara H (2008) Degenerative alterations in the visual pathway after NMDA-induced retinal damage in mice. Brain Res 1212:89–101. https://doi.org/10.1016/j.brainres.2008.03.021
Joly S, Lange C, Thiersch M, Samardzija M, Grimm C (2008) Leukemia inhibitory factor extends the lifespan of injured photoreceptors in vivo. J Neurosci 28:13765–13774. https://doi.org/10.1523/JNEUROSCI.5114-08.2008
Kaindl AM et al (2012) Activation of microglial N-methyl-d-aspartate receptors triggers inflammation and neuronal cell death in the develo** and mature brain. Ann Neurol 72:536–549. https://doi.org/10.1002/ana.23626
Kang HJ, Menlove K, Ma J, Wilkins A, Lichtarge O, Wensel TG (2014) Selectivity and evolutionary divergence of metabotropic glutamate receptors for endogenous ligands and G proteins coupled to phospholipase C or TRP channels. J Biol Chem 289:29961–29974. https://doi.org/10.1074/jbc.M114.574483
Kuribayashi J, Kitaoka Y, Munemasa Y, Ueno S (2010) Kinesin-1 and degenerative changes in optic nerve axons in NMDA-induced neurotoxicity. Brain Res 1362:133–140. https://doi.org/10.1016/j.brainres.2010.09.053
Lam TT, Abler AS, Kwong JM, Tso MO (1999) N-Methyl-d-aspartate (NMDA)-induced apoptosis in rat retina. Invest Ophthalmol Vis Sci 40:2391–2397
Mabuchi F, Aihara M, Mackey MR, Lindsey JD, Weinreb RN (2003) Optic nerve damage in experimental mouse ocular hypertension. Invest Ophthalmol Vis Sci 44:4321–4330
Manabe S, Lipton SA (2003) Divergent NMDA signals leading to proapoptotic and antiapoptotic pathways in the rat retina. Invest Ophthalmol Vis Sci 44:385–392
Meredith GE, Totterdell S, Beales M, Meshul CK (2009) Impaired glutamate homeostasis and programmed cell death in a chronic MPTP mouse model of Parkinson's disease. Exp Neurol 219:334–340. https://doi.org/10.1016/j.expneurol.2009.06.005
Mota SI, Ferreira IL, Rego AC (2014) Dysfunctional synapse in Alzheimer’s disease—a focus on NMDA receptors. Neuropharmacology 76 Pt A:16–26. https://doi.org/10.1016/j.neuropharm.2013.08.013
Nair P, Lu M, Petersen S, Ashkenazi A (2014) Apoptosis initiation through the cell-extrinsic pathway. Methods Enzymol 544:99–128. https://doi.org/10.1016/B978-0-12-417158-9.00005-4
Nakano N, Ikeda HO, Hangai M, Muraoka Y, Toda Y, Kakizuka A, Yoshimura N (2011) Longitudinal and simultaneous imaging of retinal ganglion cells and inner retinal layers in a mouse model of glaucoma induced by N-methyl-d-aspartate. Invest Ophthalmol Vis Sci 52:8754–8762. https://doi.org/10.1167/iovs.10-6654
Naruoka T et al (2013) ISO-1, a macrophage migration inhibitory factor antagonist, prevents N-methyl-d-aspartate-induced retinal damage. Eur J Pharmacol 718:138–144. https://doi.org/10.1016/j.ejphar.2013.08.041
Naskar R, Wissing M, Thanos S (2002) Detection of early neuron degeneration and accompanying microglial responses in the retina of a rat model of glaucoma. Invest Ophthalmol Vis Sci 43:2962–2968
Newsholme P, Procopio J, Lima MM, Pithon-Curi TC, Curi R (2003) Glutamine and glutamate—their central role in cell metabolism and function. Cell Biochem Funct 21:1–9. https://doi.org/10.1002/cbf.1003
Noristani R, Kuehn S, Stute G, Reinehr S, Stellbogen M, Dick HB, Joachim SC (2016) Retinal and optic nerve damage is associated with early glial responses in an experimental autoimmune glaucoma model. J Mol Neurosci 58:470–482. https://doi.org/10.1007/s12031-015-0707-2
Ohno Y, Nakanishi T, Umigai N, Tsuruma K, Shimazawa M, Hara H (2012) Oral administration of crocetin prevents inner retinal damage induced by N-methyl-d-aspartate in mice. Eur J Pharmacol 690:84–89. https://doi.org/10.1016/j.ejphar.2012.06.035
Oikawa F, Nakahara T, Akanuma K, Ueda K, Mori A, Sakamoto K, Ishii K (2012) Protective effects of the beta3-adrenoceptor agonist CL316243 against N-methyl-d-aspartate-induced retinal neurotoxicity. Naunyn Schmiedebergs Arch Pharmacol 385:1077–1081. https://doi.org/10.1007/s00210-012-0796-1
Reinehr S et al (2016) Simultaneous complement response via lectin pathway in retina and optic nerve in an experimental autoimmune glaucoma model. Front Cell Neurosci 10:140. https://doi.org/10.3389/fncel.2016.00140
Saggu SK, Chotaliya HP, Blumbergs PC, Casson RJ (2010) Wallerian-like axonal degeneration in the optic nerve after excitotoxic retinal insult: an ultrastructural study. BMC Neurosci 11:97. https://doi.org/10.1186/1471-2202-11-97
Salter MG, Fern R (2005) NMDA receptors are expressed in develo** oligodendrocyte processes and mediate injury. Nature 438:1167–1171. https://doi.org/10.1038/nature04301
Schmid H, Renner M, Dick HB, Joachim SC (2014) Loss of inner retinal neurons after retinal ischemia in rats. Invest Ophthalmol Vis Sci 55:2777–2787. https://doi.org/10.1167/iovs.13-13372
Seitz R, Tamm ER (2013) N-Methyl-d-aspartate (NMDA)-mediated excitotoxic damage: a mouse model of acute retinal ganglion cell damage. Methods Mol Biol 935:99–109. https://doi.org/10.1007/978-1-62703-080-9_7
Seitz R, Tamm ER (2014) Muller cells and microglia of the mouse eye react throughout the entire retina in response to the procedure of an intravitreal injection. Adv Exp Med Biol 801:347–353. https://doi.org/10.1007/978-1-4614-3209-8_44
Shimazawa M, Miwa A, Ito Y, Tsuruma K, Aihara M, Hara H (2012) Involvement of endoplasmic reticulum stress in optic nerve degeneration following N-methyl-d-aspartate-induced retinal damage in mice. J Neurosci Res 90:1960–1969. https://doi.org/10.1002/jnr.23078
Singhal S, Lawrence JM, Salt TE, Khaw PT, Limb GA (2010) Triamcinolone attenuates macrophage/microglia accumulation associated with NMDA-induced RGC death and facilitates survival of Muller stem cell grafts. Exp Eye Res 90:308–315. https://doi.org/10.1016/j.exer.2009.11.008
Son JL, Soto I, Oglesby E, Lopez-Roca T, Pease ME, Quigley HA, Marsh-Armstrong N (2010) Glaucomatous optic nerve injury involves early astrocyte reactivity and late oligodendrocyte loss. Glia 58:780–789. https://doi.org/10.1002/glia.20962
Susin SA et al (1999) Molecular characterization of mitochondrial apoptosis-inducing factor. Nature 397:441–446. https://doi.org/10.1038/17135
Urushitani M et al (2001) N-Methyl-d-aspartate receptor-mediated mitochondrial Ca(2+) overload in acute excitotoxic motor neuron death: a mechanism distinct from chronic neurotoxicity after Ca(2+) influx. J Neurosci Res 63:377–387
Wada Y et al (2013) PACAP attenuates NMDA-induced retinal damage in association with modulation of the microglia/macrophage status into an acquired deactivation subtype. J Mol Neurosci. https://doi.org/10.1007/s12031-013-0017-5
Wang H et al (2004) Apoptosis-inducing factor substitutes for caspase executioners in NMDA-triggered excitotoxic neuronal death. J Neurosci 24:10963–10973. https://doi.org/10.1523/JNEUROSCI.3461-04.2004
Wang X, Haroon F, Karray S, Martina D, Schluter D (2013) Astrocytic Fas ligand expression is required to induce T-cell apoptosis and recovery from experimental autoimmune encephalomyelitis. Eur J Immunol 43:115–124. https://doi.org/10.1002/eji.201242679
Wang X, Ng YK, Tay SS (2005) Factors contributing to neuronal degeneration in retinas of experimental glaucomatous rats. J Neurosci Res 82:674–689. https://doi.org/10.1002/jnr.20679
Wax MB, Tezel G (2009) Immunoregulation of retinal ganglion cell fate in glaucoma. Exp Eye Res 88:825–830. https://doi.org/10.1016/j.exer.2009.02.005
Yu SW, Andrabi SA, Wang H, Kim NS, Poirier GG, Dawson TM, Dawson VL (2006) Apoptosis-inducing factor mediates poly(ADP-ribose) (PAR) polymer-induced cell death. Proc Natl Acad Sci U S A 103:18314–18319. https://doi.org/10.1073/pnas.0606528103
Zhou Y, Tencerova B, Hartveit E, Veruki ML (2016) Functional NMDA receptors are expressed by both AII and A17 amacrine cells in the rod pathway of the mammalian retina. J Neurophysiol 115:389–403. https://doi.org/10.1152/jn.00947.2015
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kuehn, S., Rodust, C., Stute, G. et al. Concentration-Dependent Inner Retina Layer Damage and Optic Nerve Degeneration in a NMDA Model. J Mol Neurosci 63, 283–299 (2017). https://doi.org/10.1007/s12031-017-0978-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12031-017-0978-x