Retinal glutamate transporter changes in experimental glaucoma and after optic nerve transection in the rat

Keith R G Martin, Hana Levkovitch-Verbin, Danielle Valenta, Lisa Baumrind, Mary Ellen Pease, Harry A Quigley

Research output: Contribution to journalArticle

Abstract

PURPOSE. High levels of glutamate can be toxic to retinal ganglion cells, Effective buffering of extracellular glutamate by retinal glutamate transporters is therefore important, This study was conducted to investigate whether glutamate transporter changes occur with two models of optic nerve injury in the rat. METHODS. Glaucoma was induced in one eye of 35 adult Wistar rats by translimbal diode laser treatment to the trabecular meshwork. Twenty-five more rats underwent unilateral optic nerve transection. Two glutamate transporters, GLAST (EAAT-1) and GLT-1 (EAAT-2), were studied by immunohistochemistry and quantitative Western blot analysis. Treated and control eyes were compared 3 days and 1, 4, and 6 weeks after injury. Optic nerve damage was assessed semiquantitatively in epoxy-embedded optic nerve cross sections. RESULTS. Trabecular laser treatment resulted in moderate intraocular pressure (IOP) elevation in all animals. After 1 to 6 weeks of experimental glaucoma, all treated eyes had significant optic nerve damage. Glutamate transporter changes were not detected by immunohistochemistry. Western blot analysis demonstrated significantly reduced GLT-1 in glaucomatous eyes compared with control eyes at 3 days (29.3% ± 6.7%, P = 0.01), 1 week (55.5% ± 13.6%, P = 0.02), 4 weeks (27.2% ± 10.1%, P = 0.05), and 6 weeks (38.1% ± 7,9%, P = 0.01; mean reduction ± SEM, paired t-tests, n = 5 animals per group, four duplicate Western blot analyses per eye). The magnitude of the reduction in GLT-1 correlated significantly with mean IOP in the glaucomatous eye (r2 = 0.31, P = 0.01, linear regression). GLAST was significantly reduced (33.8% ± 8.1%, mean ± SEM) after 4 weeks of elevated IOP (P = 0.01, paired t-test, n = 5 animals per group). In contrast to glaucoma, optic nerve transection resulted in an increase in GLT-1 compared with the control eye (P = 0.01, paired t-test, n = 15 animals). There was no significant change in GLAST after transection. CONCLUSIONS. GLT-1 and GLAST were significantly reduced in an experimental rat glaucoma model, a response that was not found after optic nerve transection. Reductions in GLT-1 and GLAST may increase the potential for glutamate-induced injury to RGC in glaucoma.

Original languageEnglish (US)
Pages (from-to)2236-2243
Number of pages8
JournalInvestigative Ophthalmology and Visual Science
Volume43
Issue number7
StatePublished - 2002

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Optic Nerve Injuries
Amino Acid Transport System X-AG
Glaucoma
Optic Nerve
Intraocular Pressure
Glutamic Acid
Western Blotting
Immunohistochemistry
Trabecular Meshwork
Semiconductor Lasers
Retinal Ganglion Cells
Poisons
Wounds and Injuries
Wistar Rats
Linear Models
Lasers

ASJC Scopus subject areas

  • Ophthalmology

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Retinal glutamate transporter changes in experimental glaucoma and after optic nerve transection in the rat. / Martin, Keith R G; Levkovitch-Verbin, Hana; Valenta, Danielle; Baumrind, Lisa; Pease, Mary Ellen; Quigley, Harry A.

In: Investigative Ophthalmology and Visual Science, Vol. 43, No. 7, 2002, p. 2236-2243.

Research output: Contribution to journalArticle

Martin, Keith R G ; Levkovitch-Verbin, Hana ; Valenta, Danielle ; Baumrind, Lisa ; Pease, Mary Ellen ; Quigley, Harry A. / Retinal glutamate transporter changes in experimental glaucoma and after optic nerve transection in the rat. In: Investigative Ophthalmology and Visual Science. 2002 ; Vol. 43, No. 7. pp. 2236-2243.
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abstract = "PURPOSE. High levels of glutamate can be toxic to retinal ganglion cells, Effective buffering of extracellular glutamate by retinal glutamate transporters is therefore important, This study was conducted to investigate whether glutamate transporter changes occur with two models of optic nerve injury in the rat. METHODS. Glaucoma was induced in one eye of 35 adult Wistar rats by translimbal diode laser treatment to the trabecular meshwork. Twenty-five more rats underwent unilateral optic nerve transection. Two glutamate transporters, GLAST (EAAT-1) and GLT-1 (EAAT-2), were studied by immunohistochemistry and quantitative Western blot analysis. Treated and control eyes were compared 3 days and 1, 4, and 6 weeks after injury. Optic nerve damage was assessed semiquantitatively in epoxy-embedded optic nerve cross sections. RESULTS. Trabecular laser treatment resulted in moderate intraocular pressure (IOP) elevation in all animals. After 1 to 6 weeks of experimental glaucoma, all treated eyes had significant optic nerve damage. Glutamate transporter changes were not detected by immunohistochemistry. Western blot analysis demonstrated significantly reduced GLT-1 in glaucomatous eyes compared with control eyes at 3 days (29.3{\%} ± 6.7{\%}, P = 0.01), 1 week (55.5{\%} ± 13.6{\%}, P = 0.02), 4 weeks (27.2{\%} ± 10.1{\%}, P = 0.05), and 6 weeks (38.1{\%} ± 7,9{\%}, P = 0.01; mean reduction ± SEM, paired t-tests, n = 5 animals per group, four duplicate Western blot analyses per eye). The magnitude of the reduction in GLT-1 correlated significantly with mean IOP in the glaucomatous eye (r2 = 0.31, P = 0.01, linear regression). GLAST was significantly reduced (33.8{\%} ± 8.1{\%}, mean ± SEM) after 4 weeks of elevated IOP (P = 0.01, paired t-test, n = 5 animals per group). In contrast to glaucoma, optic nerve transection resulted in an increase in GLT-1 compared with the control eye (P = 0.01, paired t-test, n = 15 animals). There was no significant change in GLAST after transection. CONCLUSIONS. GLT-1 and GLAST were significantly reduced in an experimental rat glaucoma model, a response that was not found after optic nerve transection. Reductions in GLT-1 and GLAST may increase the potential for glutamate-induced injury to RGC in glaucoma.",
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T1 - Retinal glutamate transporter changes in experimental glaucoma and after optic nerve transection in the rat

AU - Martin, Keith R G

AU - Levkovitch-Verbin, Hana

AU - Valenta, Danielle

AU - Baumrind, Lisa

AU - Pease, Mary Ellen

AU - Quigley, Harry A

PY - 2002

Y1 - 2002

N2 - PURPOSE. High levels of glutamate can be toxic to retinal ganglion cells, Effective buffering of extracellular glutamate by retinal glutamate transporters is therefore important, This study was conducted to investigate whether glutamate transporter changes occur with two models of optic nerve injury in the rat. METHODS. Glaucoma was induced in one eye of 35 adult Wistar rats by translimbal diode laser treatment to the trabecular meshwork. Twenty-five more rats underwent unilateral optic nerve transection. Two glutamate transporters, GLAST (EAAT-1) and GLT-1 (EAAT-2), were studied by immunohistochemistry and quantitative Western blot analysis. Treated and control eyes were compared 3 days and 1, 4, and 6 weeks after injury. Optic nerve damage was assessed semiquantitatively in epoxy-embedded optic nerve cross sections. RESULTS. Trabecular laser treatment resulted in moderate intraocular pressure (IOP) elevation in all animals. After 1 to 6 weeks of experimental glaucoma, all treated eyes had significant optic nerve damage. Glutamate transporter changes were not detected by immunohistochemistry. Western blot analysis demonstrated significantly reduced GLT-1 in glaucomatous eyes compared with control eyes at 3 days (29.3% ± 6.7%, P = 0.01), 1 week (55.5% ± 13.6%, P = 0.02), 4 weeks (27.2% ± 10.1%, P = 0.05), and 6 weeks (38.1% ± 7,9%, P = 0.01; mean reduction ± SEM, paired t-tests, n = 5 animals per group, four duplicate Western blot analyses per eye). The magnitude of the reduction in GLT-1 correlated significantly with mean IOP in the glaucomatous eye (r2 = 0.31, P = 0.01, linear regression). GLAST was significantly reduced (33.8% ± 8.1%, mean ± SEM) after 4 weeks of elevated IOP (P = 0.01, paired t-test, n = 5 animals per group). In contrast to glaucoma, optic nerve transection resulted in an increase in GLT-1 compared with the control eye (P = 0.01, paired t-test, n = 15 animals). There was no significant change in GLAST after transection. CONCLUSIONS. GLT-1 and GLAST were significantly reduced in an experimental rat glaucoma model, a response that was not found after optic nerve transection. Reductions in GLT-1 and GLAST may increase the potential for glutamate-induced injury to RGC in glaucoma.

AB - PURPOSE. High levels of glutamate can be toxic to retinal ganglion cells, Effective buffering of extracellular glutamate by retinal glutamate transporters is therefore important, This study was conducted to investigate whether glutamate transporter changes occur with two models of optic nerve injury in the rat. METHODS. Glaucoma was induced in one eye of 35 adult Wistar rats by translimbal diode laser treatment to the trabecular meshwork. Twenty-five more rats underwent unilateral optic nerve transection. Two glutamate transporters, GLAST (EAAT-1) and GLT-1 (EAAT-2), were studied by immunohistochemistry and quantitative Western blot analysis. Treated and control eyes were compared 3 days and 1, 4, and 6 weeks after injury. Optic nerve damage was assessed semiquantitatively in epoxy-embedded optic nerve cross sections. RESULTS. Trabecular laser treatment resulted in moderate intraocular pressure (IOP) elevation in all animals. After 1 to 6 weeks of experimental glaucoma, all treated eyes had significant optic nerve damage. Glutamate transporter changes were not detected by immunohistochemistry. Western blot analysis demonstrated significantly reduced GLT-1 in glaucomatous eyes compared with control eyes at 3 days (29.3% ± 6.7%, P = 0.01), 1 week (55.5% ± 13.6%, P = 0.02), 4 weeks (27.2% ± 10.1%, P = 0.05), and 6 weeks (38.1% ± 7,9%, P = 0.01; mean reduction ± SEM, paired t-tests, n = 5 animals per group, four duplicate Western blot analyses per eye). The magnitude of the reduction in GLT-1 correlated significantly with mean IOP in the glaucomatous eye (r2 = 0.31, P = 0.01, linear regression). GLAST was significantly reduced (33.8% ± 8.1%, mean ± SEM) after 4 weeks of elevated IOP (P = 0.01, paired t-test, n = 5 animals per group). In contrast to glaucoma, optic nerve transection resulted in an increase in GLT-1 compared with the control eye (P = 0.01, paired t-test, n = 15 animals). There was no significant change in GLAST after transection. CONCLUSIONS. GLT-1 and GLAST were significantly reduced in an experimental rat glaucoma model, a response that was not found after optic nerve transection. Reductions in GLT-1 and GLAST may increase the potential for glutamate-induced injury to RGC in glaucoma.

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