Skip to main content
PMC home page
Transactions of the American Ophthalmological Society logoLink to Transactions of the American Ophthalmological Society
. 2003;101:39–51.

Retinal ganglion cell protection with geranylgeranylacetone, a heat shock protein inducer, in a rat glaucoma model.

Joseph Caprioli 1, Yoko Ishii 1, Jacky M K Kwong 1
PMCID: PMC1358973  PMID: 14971562

Abstract

PURPOSE: To study the effects of geranylgeranylacetone (GCA) on the expression of inducible (HSP72) and constitutive (HSC70) heat shock proteins (HSPs) on retinal ganglion cells (RGCs) in a rat model of glaucoma. METHODS: Adult Wistar rats were given intraperitoneal injections of GGA, 200 mg/kg daily. Western blot analysis and immunohistochemical staining for HSP72 and HSC70 were performed after 1, 3, and 7 days of GGA administration. After 7 days of GGA pretreatment, intraocular pressure (IOP) was elevated unilaterally by repeated trabecular argon laser photocoagulation 5 days after intracameral injection of india ink. After the first laser photocoagulation, CGA was given twice a week. RGC survival was evaluated after 5 weeks of IOP elevation. Immunohistochemistry and TdT-mediated biotin-dUTP nick end labeling (TUNEL) were performed after 1 week of IOP elevation. Quercetin, an inhibitor of HSP expression, was also administered to a separate group. RESULTS: There was increased expression of HSP72 in RGCs at 3 and 7 days after GGA administration, but HSC70 was unchanged. After 5 weeks of IOP elevation, there was 27% +/- 6% loss of RGCs. The administration of GGA significantly reduced the loss of RGCs, lessened optic nerve damage, decreased the number of TUNEL-positive cells in the RGC layer, and increased HSP72. Quercetin administration abolished these protective effects. CONCLUSIONS: These results demonstrate that systemic administration of GGA protects RGCs from glaucomatous damage in a rat model and suggest a novel pathway for netroprotection for patients with glaucoma.

Full Text

The Full Text of this article is available as a PDF (263.3 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alavez S., Pedroza D., Morán J. Role of heat shock proteins in the effect of NMDA and KCl on cerebellar granule cells survival. Neurochem Res. 2000 Mar;25(3):341–347. doi: 10.1023/a:1007584802989. [DOI] [PubMed] [Google Scholar]
  2. Barbe M. F., Tytell M., Gower D. J., Welch W. J. Hyperthermia protects against light damage in the rat retina. Science. 1988 Sep 30;241(4874):1817–1820. doi: 10.1126/science.3175623. [DOI] [PubMed] [Google Scholar]
  3. Beere H. M., Green D. R. Stress management - heat shock protein-70 and the regulation of apoptosis. Trends Cell Biol. 2001 Jan;11(1):6–10. doi: 10.1016/s0962-8924(00)01874-2. [DOI] [PubMed] [Google Scholar]
  4. Brown I. R. Induction of heat shock (stress) genes in the mammalian brain by hyperthermia and other traumatic events: a current perspective. J Neurosci Res. 1990 Nov;27(3):247–255. doi: 10.1002/jnr.490270302. [DOI] [PubMed] [Google Scholar]
  5. Buzzard K. A., Giaccia A. J., Killender M., Anderson R. L. Heat shock protein 72 modulates pathways of stress-induced apoptosis. J Biol Chem. 1998 Jul 3;273(27):17147–17153. doi: 10.1074/jbc.273.27.17147. [DOI] [PubMed] [Google Scholar]
  6. Caprioli J., Kitano S., Morgan J. E. Hyperthermia and hypoxia increase tolerance of retinal ganglion cells to anoxia and excitotoxicity. Invest Ophthalmol Vis Sci. 1996 Nov;37(12):2376–2381. [PubMed] [Google Scholar]
  7. Chopp M., Tidwell C. D., Lee Y. J., Knight R., Helpern J. A., Welch K. M. Reduction of hyperthermic ischemic acidosis by a conditioning event in cats. Stroke. 1989 Oct;20(10):1357–1360. doi: 10.1161/01.str.20.10.1357. [DOI] [PubMed] [Google Scholar]
  8. Feldman D. E., Frydman J. Protein folding in vivo: the importance of molecular chaperones. Curr Opin Struct Biol. 2000 Feb;10(1):26–33. doi: 10.1016/s0959-440x(99)00044-5. [DOI] [PubMed] [Google Scholar]
  9. Fink A. L. Chaperone-mediated protein folding. Physiol Rev. 1999 Apr;79(2):425–449. doi: 10.1152/physrev.1999.79.2.425. [DOI] [PubMed] [Google Scholar]
  10. Fudaba Y., Ohdan H., Tashiro H., Ito H., Fukuda Y., Dohi K., Asahara T. Geranylgeranylacetone, a heat shock protein inducer, prevents primary graft nonfunction in rat liver transplantation. Transplantation. 2001 Jul 27;72(2):184–189. doi: 10.1097/00007890-200107270-00003. [DOI] [PubMed] [Google Scholar]
  11. Gellrich M. M., Gellrich N. C. Quantitative relations in the retinal ganglion cell layer of the rat: neurons, glia and capillaries before and after optic nerve section. Graefes Arch Clin Exp Ophthalmol. 1996 May;234(5):315–323. doi: 10.1007/BF00220707. [DOI] [PubMed] [Google Scholar]
  12. Glovinsky Y., Quigley H. A., Dunkelberger G. R. Retinal ganglion cell loss is size dependent in experimental glaucoma. Invest Ophthalmol Vis Sci. 1991 Mar;32(3):484–491. [PubMed] [Google Scholar]
  13. Hirakawa T., Rokutan K., Nikawa T., Kishi K. Geranylgeranylacetone induces heat shock proteins in cultured guinea pig gastric mucosal cells and rat gastric mucosa. Gastroenterology. 1996 Aug;111(2):345–357. doi: 10.1053/gast.1996.v111.pm8690199. [DOI] [PubMed] [Google Scholar]
  14. Ikeyama S., Kusumoto K., Miyake H., Rokutan K., Tashiro S. A non-toxic heat shock protein 70 inducer, geranylgeranylacetone, suppresses apoptosis of cultured rat hepatocytes caused by hydrogen peroxide and ethanol. J Hepatol. 2001 Jul;35(1):53–61. doi: 10.1016/s0168-8278(01)00053-8. [DOI] [PubMed] [Google Scholar]
  15. Jia L., Cepurna W. O., Johnson E. C., Morrison J. C. Effect of general anesthetics on IOP in rats with experimental aqueous outflow obstruction. Invest Ophthalmol Vis Sci. 2000 Oct;41(11):3415–3419. [PubMed] [Google Scholar]
  16. Jia L., Cepurna W. O., Johnson E. C., Morrison J. C. Patterns of intraocular pressure elevation after aqueous humor outflow obstruction in rats. Invest Ophthalmol Vis Sci. 2000 May;41(6):1380–1385. [PubMed] [Google Scholar]
  17. Jättelä M., Wissing D., Kokholm K., Kallunki T., Egeblad M. Hsp70 exerts its anti-apoptotic function downstream of caspase-3-like proteases. EMBO J. 1998 Nov 2;17(21):6124–6134. doi: 10.1093/emboj/17.21.6124. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kerrigan L. A., Zack D. J., Quigley H. A., Smith S. D., Pease M. E. TUNEL-positive ganglion cells in human primary open-angle glaucoma. Arch Ophthalmol. 1997 Aug;115(8):1031–1035. doi: 10.1001/archopht.1997.01100160201010. [DOI] [PubMed] [Google Scholar]
  19. Kitagawa K., Matsumoto M., Kuwabara K., Tagaya M., Ohtsuki T., Hata R., Ueda H., Handa N., Kimura K., Kamada T. 'Ischemic tolerance' phenomenon detected in various brain regions. Brain Res. 1991 Oct 11;561(2):203–211. doi: 10.1016/0006-8993(91)91596-s. [DOI] [PubMed] [Google Scholar]
  20. Kurihara T., Kitamura Y., Adachi Y., Obuchi M., Abe K., Akimoto M., Hashimoto H., Ishiguro H., Niimi A., Maeda A. Increase in hepatic tissue blood flow by teprenone. J Gastroenterol Hepatol. 1996 Oct;11(10):978–984. doi: 10.1111/j.1440-1746.1996.tb01857.x. [DOI] [PubMed] [Google Scholar]
  21. Lee J. E., Yenari M. A., Sun G. H., Xu L., Emond M. R., Cheng D., Steinberg G. K., Giffard R. G. Differential neuroprotection from human heat shock protein 70 overexpression in in vitro and in vivo models of ischemia and ischemia-like conditions. Exp Neurol. 2001 Jul;170(1):129–139. doi: 10.1006/exnr.2000.7614. [DOI] [PubMed] [Google Scholar]
  22. Li C. Y., Lee J. S., Ko Y. G., Kim J. I., Seo J. S. Heat shock protein 70 inhibits apoptosis downstream of cytochrome c release and upstream of caspase-3 activation. J Biol Chem. 2000 Aug 18;275(33):25665–25671. doi: 10.1074/jbc.M906383199. [DOI] [PubMed] [Google Scholar]
  23. Macario A. J., Conway de Macario E. Stress and molecular chaperones in disease. Int J Clin Lab Res. 2000;30(2):49–66. doi: 10.1007/s005990070016. [DOI] [PubMed] [Google Scholar]
  24. Mailhos C., Howard M. K., Latchman D. S. Heat shock protects neuronal cells from programmed cell death by apoptosis. Neuroscience. 1993 Aug;55(3):621–627. doi: 10.1016/0306-4522(93)90428-i. [DOI] [PubMed] [Google Scholar]
  25. Miwa H., Nagahara A., Wada R., Hirai S., Endo K., Watanabe S., Sato N. Suppressive effect of tetraprenylacetone on gastric atrophy induced by short-term administration of N-methyl-N'-nitro-N-nitrosoguanidine in rats. Aliment Pharmacol Ther. 1997 Jun;11(3):613–618. doi: 10.1046/j.1365-2036.1997.00162.x. [DOI] [PubMed] [Google Scholar]
  26. Mizushima T., Tsutsumi S., Rokutan K., Tsuchiya T. Suppression of ethanol-induced apoptotic DNA fragmentation by geranylgeranylacetone in cultured guinea pig gastric mucosal cells. Dig Dis Sci. 1999 Mar;44(3):510–514. doi: 10.1023/a:1026692920848. [DOI] [PubMed] [Google Scholar]
  27. Morimoto R. I. Cells in stress: transcriptional activation of heat shock genes. Science. 1993 Mar 5;259(5100):1409–1410. doi: 10.1126/science.8451637. [DOI] [PubMed] [Google Scholar]
  28. Morimoto R. I., Santoro M. G. Stress-inducible responses and heat shock proteins: new pharmacologic targets for cytoprotection. Nat Biotechnol. 1998 Sep;16(9):833–838. doi: 10.1038/nbt0998-833. [DOI] [PubMed] [Google Scholar]
  29. Mosser D. D., Caron A. W., Bourget L., Denis-Larose C., Massie B. Role of the human heat shock protein hsp70 in protection against stress-induced apoptosis. Mol Cell Biol. 1997 Sep;17(9):5317–5327. doi: 10.1128/mcb.17.9.5317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Mosser D. D., Caron A. W., Bourget L., Meriin A. B., Sherman M. Y., Morimoto R. I., Massie B. The chaperone function of hsp70 is required for protection against stress-induced apoptosis. Mol Cell Biol. 2000 Oct;20(19):7146–7159. doi: 10.1128/mcb.20.19.7146-7159.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Murakami M., Oketani K., Fujisaki H., Wakabayashi T., Ohgo T. Antiulcer effect of geranylgeranylacetone, a new acyclic polyisoprenoid on experimentally induced gastric and duodenal ulcers in rats. Arzneimittelforschung. 1981;31(5):799–804. [PubMed] [Google Scholar]
  32. Nishida K., Ohta Y., Ishiguro I. Preventive effect of teprenone on stress-induced gastric mucosal lesions and its relation to gastric mucosal constitutive nitric oxide synthase activity. Pharmacol Res. 1999 Apr;39(4):325–332. doi: 10.1006/phrs.1998.0448. [DOI] [PubMed] [Google Scholar]
  33. Nylandsted J., Rohde M., Brand K., Bastholm L., Elling F., Jättelä M. Selective depletion of heat shock protein 70 (Hsp70) activates a tumor-specific death program that is independent of caspases and bypasses Bcl-2. Proc Natl Acad Sci U S A. 2000 Jul 5;97(14):7871–7876. doi: 10.1073/pnas.97.14.7871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Okisaka S., Murakami A., Mizukawa A., Ito J. Apoptosis in retinal ganglion cell decrease in human glaucomatous eyes. Jpn J Ophthalmol. 1997 Mar-Apr;41(2):84–88. doi: 10.1016/s0021-5155(97)00013-0. [DOI] [PubMed] [Google Scholar]
  35. Ooie T., Takahashi N., Saikawa T., Nawata T., Arikawa M., Yamanaka K., Hara M., Shimada T., Sakata T. Single oral dose of geranylgeranylacetone induces heat-shock protein 72 and renders protection against ischemia/reperfusion injury in rat heart. Circulation. 2001 Oct 9;104(15):1837–1843. doi: 10.1161/hc3901.095771. [DOI] [PubMed] [Google Scholar]
  36. Park K. H., Cozier F., Ong O. C., Caprioli J. Induction of heat shock protein 72 protects retinal ganglion cells in a rat glaucoma model. Invest Ophthalmol Vis Sci. 2001 Jun;42(7):1522–1530. [PubMed] [Google Scholar]
  37. Plumier J. C., Krueger A. M., Currie R. W., Kontoyiannis D., Kollias G., Pagoulatos G. N. Transgenic mice expressing the human inducible Hsp70 have hippocampal neurons resistant to ischemic injury. Cell Stress Chaperones. 1997 Sep;2(3):162–167. doi: 10.1379/1466-1268(1997)002<0162:tmethi>2.3.co;2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Quigley H. A., Dunkelberger G. R., Green W. R. Chronic human glaucoma causing selectively greater loss of large optic nerve fibers. Ophthalmology. 1988 Mar;95(3):357–363. doi: 10.1016/s0161-6420(88)33176-3. [DOI] [PubMed] [Google Scholar]
  39. Quigley H. A., Nickells R. W., Kerrigan L. A., Pease M. E., Thibault D. J., Zack D. J. Retinal ganglion cell death in experimental glaucoma and after axotomy occurs by apoptosis. Invest Ophthalmol Vis Sci. 1995 Apr;36(5):774–786. [PubMed] [Google Scholar]
  40. Rokutan K., Hirakawa T., Teshima S., Nakano Y., Miyoshi M., Kawai T., Konda E., Morinaga H., Nikawa T., Kishi K. Implications of heat shock/stress proteins for medicine and disease. J Med Invest. 1998 Feb;44(3-4):137–147. [PubMed] [Google Scholar]
  41. Rokutan K., Teshima S., Kawai T., Kawahara T., Kusumoro K., Mizushima T., Kishi K. Geranylgeranylacetone stimulates mucin synthesis in cultured guinea pig gastric pit cells by inducing a neuronal nitric oxide synthase. J Gastroenterol. 2000;35(9):673–681. doi: 10.1007/s005350070046. [DOI] [PubMed] [Google Scholar]
  42. Rordorf G., Koroshetz W. J., Bonventre J. V. Heat shock protects cultured neurons from glutamate toxicity. Neuron. 1991 Dec;7(6):1043–1051. doi: 10.1016/0896-6273(91)90348-4. [DOI] [PubMed] [Google Scholar]
  43. Saleh A., Srinivasula S. M., Balkir L., Robbins P. D., Alnemri E. S. Negative regulation of the Apaf-1 apoptosome by Hsp70. Nat Cell Biol. 2000 Aug;2(8):476–483. doi: 10.1038/35019510. [DOI] [PubMed] [Google Scholar]
  44. Santoro M. G. Heat shock factors and the control of the stress response. Biochem Pharmacol. 2000 Jan 1;59(1):55–63. doi: 10.1016/s0006-2952(99)00299-3. [DOI] [PubMed] [Google Scholar]
  45. Sommer A., Katz J., Quigley H. A., Miller N. R., Robin A. L., Richter R. C., Witt K. A. Clinically detectable nerve fiber atrophy precedes the onset of glaucomatous field loss. Arch Ophthalmol. 1991 Jan;109(1):77–83. doi: 10.1001/archopht.1991.01080010079037. [DOI] [PubMed] [Google Scholar]
  46. Stacchiotti A., Rezzani R., Rodella L., Tiberio L., Schiaffonati L., Bianchi R. Cell-specific expression of heat shock transcription factors 1 and 2 in unstressed rat spinal cord. Neurosci Lett. 1999 Jun 18;268(2):73–76. doi: 10.1016/s0304-3940(99)00394-8. [DOI] [PubMed] [Google Scholar]
  47. Takahashi S., Yamazaki T., Okabe S. Effects of heat shock and teprenone on ethanol-induced damage to cultured rabbit gastric mucosal cells. J Physiol Pharmacol. 1996 Sep;47(3):433–441. [PubMed] [Google Scholar]
  48. Tatton N. A., Tezel G., Insolia S. A., Nandor S. A., Edward P. D., Wax M. B. In situ detection of apoptosis in normal pressure glaucoma. a preliminary examination. Surv Ophthalmol. 2001 May;45 (Suppl 3):S268–S276. doi: 10.1016/s0039-6257(01)00204-1. [DOI] [PubMed] [Google Scholar]
  49. Tezel G., Hernandez R., Wax M. B. Immunostaining of heat shock proteins in the retina and optic nerve head of normal and glaucomatous eyes. Arch Ophthalmol. 2000 Apr;118(4):511–518. doi: 10.1001/archopht.118.4.511. [DOI] [PubMed] [Google Scholar]
  50. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Tsuruma T., Yagihashi A., Hirata K., Araya J., Katsuramaki T., Tarumi K., Yanai Y., Watanabe N. Induction of heat shock protein-70 (hsp-70) by intraarterial administration of geranylgeranylacetone. Transplant Proc. 2000 Nov;32(7):1631–1633. doi: 10.1016/s0041-1345(00)01441-x. [DOI] [PubMed] [Google Scholar]
  52. Tsuruma T., Yagihashi A., Koide S., Araya J., Tarumi K., Watanabe N., Hirata K. Geranylgeranylacetone induces heat shock protein-73 in rat small intestine. Transplant Proc. 1999 Feb-Mar;31(1-2):572–573. doi: 10.1016/s0041-1345(98)01559-0. [DOI] [PubMed] [Google Scholar]
  53. Tsutsumi S., Rokutan K., Tsuchiya T., Mizushima T. Geranylgeranylacetone suppresses spontaneous apoptotic DNA fragmentation in cultured guinea pig gastric pit cells. Biol Pharm Bull. 1999 Aug;22(8):886–887. doi: 10.1248/bpb.22.886. [DOI] [PubMed] [Google Scholar]
  54. Ueda J., Sawaguchi S., Hanyu T., Yaoeda K., Fukuchi T., Abe H., Ozawa H. Experimental glaucoma model in the rat induced by laser trabecular photocoagulation after an intracameral injection of India ink. Jpn J Ophthalmol. 1998 Sep-Oct;42(5):337–344. doi: 10.1016/s0021-5155(98)00026-4. [DOI] [PubMed] [Google Scholar]
  55. Welch W. J. Mammalian stress response: cell physiology, structure/function of stress proteins, and implications for medicine and disease. Physiol Rev. 1992 Oct;72(4):1063–1081. doi: 10.1152/physrev.1992.72.4.1063. [DOI] [PubMed] [Google Scholar]
  56. Wu C. Heat shock transcription factors: structure and regulation. Annu Rev Cell Dev Biol. 1995;11:441–469. doi: 10.1146/annurev.cb.11.110195.002301. [DOI] [PubMed] [Google Scholar]
  57. Yenari M. A., Fink S. L., Sun G. H., Chang L. K., Patel M. K., Kunis D. M., Onley D., Ho D. Y., Sapolsky R. M., Steinberg G. K. Gene therapy with HSP72 is neuroprotective in rat models of stroke and epilepsy. Ann Neurol. 1998 Oct;44(4):584–591. doi: 10.1002/ana.410440403. [DOI] [PubMed] [Google Scholar]

Articles from Transactions of the American Ophthalmological Society are provided here courtesy of American Ophthalmological Society

RESOURCES