Japanese Journal of Ophthalmology

, Volume 52, Issue 3, pp 217–223 | Cite as

Effect of electrical stimulation on IGF-1 transcription by L-type calcium channels in cultured retinal Müller cells

  • Tatsuhiko Sato
  • Takashi FujikadoEmail author
  • Takeshi Morimoto
  • Kenji Matsushita
  • Takayuki Harada
  • Yasuo Tano
Laboratory Investgation



To investigate the effect of electrical stimulation (ES) on the induction of insulin-like growth factor 1 (IGF-1) in cultured retinal Müller cells.


Müller cells were isolated from rat retinas. ES was applied to Müller cells of passage 1 with biphasic pulses (duration, 1 ms; frequency, 20 Hz; current, 0–10 mA) for 30 min. The mRNA level of IGF-1 was determined by reverse transcription-polymerase chain reaction (RT-PCR) immediately to 2 h after ES. The change of intracellular calcium concentration ([Ca2+]) induced by ES was monitored by Ca2+ imaging with Fura 2-AM. Ca2+ imaging and RT-PCR were performed with and without the application of l μM nifedipine, an L-type calcium channel blocker.


The mRNA level of IGF-1 was increased significantly (P < 0.05) by about 1.3-fold immediately after 10 mA ES. [Ca2+] began to increase immediately after the start of ES, reached a maximum of approximately 1.8-fold, and continued to increase until about 20 min after the ES. The inductions of IGF-1 transcription and Ca2+ influx were suppressed by nifedipine.


These results indicate that the enhancement of IGF-1 transcription by ES in cultured Müller cells depends largely on Ca2+ influx through L-type Ca2+ channels.

Key Words

electrical stimulation IGF-1 L-type calcium channel Müller cell 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Morimoto T, Miyoshi T, Matsuda S, Tano Y, Fujikado T, Fukuda Y. Transcorneal electrical stimulation rescues axotomized retinal ganglion cells by activating endogenous retinal IGF-1 system. Invest Ophthalmol Vis Sci 2005;46:2147–2155.CrossRefPubMedGoogle Scholar
  2. 2.
    Fujikado T, Morimoto T, Matsushita K, Shimojo H, Okawa Y, Tano Y. Effect of transcorneal electrical stimulation in patients with nonarteritic ischemic optic neuropathy or traumatic optic neuropathy. Jpn J Ophthalmol 2006;50:266–273.CrossRefPubMedGoogle Scholar
  3. 3.
    Porter JT, McCarthy KD. Hippocampal astrocytes in situ respond to glutamate released from synaptic terminals. J Neurosci 1996;16:5073–5081.PubMedGoogle Scholar
  4. 4.
    Koyama S, Haruyama T, Kobatake E, Aizawa M. Electrically induced NGF production by astroglial cells. Nat Biotechnol 1997;15:164–166.CrossRefPubMedGoogle Scholar
  5. 5.
    Yanagida Y, Mizuno A, Motegi T, Kobatake E, Aizawa M. Electrically stimulated induction of hsp70 gene expression in mouse astroglia and fibroblast cells. J Biotech 2000;79:53–61.CrossRefGoogle Scholar
  6. 6.
    Schipke CG, Boucsein C, Ohlemeyer C, Kirchhoff F, Kettenmann H. Astrocyte Ca2+ waves trigger responses in microglial cells in brain slices. FASEB J 2002;16:255–257.CrossRefPubMedGoogle Scholar
  7. 7.
    Balkowiec A, Katz DM. Cellular mechanisms regulating activitydependent release of native brain-derived neurotrophic factor from hippocampal neurons. J Neurosci 2002;22:10399–10407.PubMedGoogle Scholar
  8. 8.
    Simard M, Arcuino G, Takano T, Liu QS, Nedergaard M. Signaling at the gliovascular interface. J Neurosci 2003;23:9254–9262.PubMedGoogle Scholar
  9. 9.
    Bowser DN, Khakh BS. ATP excites interneurons and astrocytes to increase synaptic inhibition in neuronal networks. J Neurosci 2004;24:8606–8620.CrossRefPubMedGoogle Scholar
  10. 10.
    Cao W, Wen R, Li F, Cheng T, Steinberg RH. Induction of basic fibroblast growth factor mRNA by basic fibroblast growth factor in Müller cells. Invest Ophthalmol Vis Sci 1997;38:1358–1366.PubMedGoogle Scholar
  11. 11.
    Li F, Cao W, Steinberg RH, LaVail MM. Basic FGF-induced down-regulation of IGF-1 mRNA in cultured rat Müller cells. Exp Eye Res 1999;68:19–27.CrossRefPubMedGoogle Scholar
  12. 12.
    Taylor S, Srinivasan B, Wordinger RJ, Roque RS. Glutamate stimulates neurotrophin expression in cultured Müller cells. Brain Res Mol Brain Res 2003;111:189–197.CrossRefPubMedGoogle Scholar
  13. 13.
    King JL, Guidry C. Insulin-like growth factor binding proteins modulate Müller cell responses to insulin-like growth factors. Invest Ophthalmol Vis Sci 2004;45:2848–2855.CrossRefPubMedGoogle Scholar
  14. 14.
    Seki M, Tanaka T, Sakai Y, et al. Müller cells as a source of brainderived neurotrophic factor in the retina: noradrenaline upregulates brain-derived neurotrophic factor levels in cultured rat Müller cells. Neurochem Res 2005;30:1163–1170.CrossRefPubMedGoogle Scholar
  15. 15.
    Roque RS, Caldwell RB, Behzadian MA. Cultured Müller cells have high levels of epidermal growth factor receptors. Invest Ophthalmol Vis Sci 1992;33:2587–2595.PubMedGoogle Scholar
  16. 16.
    Johnson MR, Wang K, Smith JB, Heslin MJ, Diasio RB. Quantitation of dihydropyrimidine dehydrogenase expression by real-time reverse transcription polymerase chain reaction. Analytical Biochem 2000;278:175–184.CrossRefGoogle Scholar
  17. 17.
    Xu HP, Zhao JW, Yang XL. Expression of voltage-dependent calcium channel subunits in the rat retina. Neurosci Lett 2002;329:297–300.CrossRefPubMedGoogle Scholar
  18. 18.
    Bourinet E, Mangoni ME, Nargeot J. Dissecting the functional role of different isoforms of the L-type Ca2+ channel. J Clin Invest 2004;113:1382–1384.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Hicks D, Courtois Y. The growth and behaviour of rat retinal Müller cells in vitro. 1. An improved method for isolation and culture. Exp Eye Res 1990;51:119–129.CrossRefPubMedGoogle Scholar
  20. 20.
    Hauck SM, Suppmann S, Ueffing M. Proteomic profiling of primary retinal Müller glia cells reveals a shift in expression patterns upon adaptation to in vitro conditions. Glia 2003;44:251–263.CrossRefPubMedGoogle Scholar
  21. 21.
    Miller RJ. Rocking and rolling with Ca2+ channel. Trends Neurosci 2001;24:445–449.CrossRefPubMedGoogle Scholar
  22. 22.
    Heidinger V, Hicks D, Sahel J, Dreyfus H. Ability of retinal Müller glial cells to protect neurons against excitotoxicity in vitro depends upon maturation and neuron-glial interactions. Glia 1999;25:229–239.CrossRefPubMedGoogle Scholar

Copyright information

© Japanese Ophthalmological Society (JOS) 2008

Authors and Affiliations

  • Tatsuhiko Sato
    • 1
  • Takashi Fujikado
    • 1
    Email author
  • Takeshi Morimoto
    • 2
  • Kenji Matsushita
    • 2
  • Takayuki Harada
    • 3
  • Yasuo Tano
    • 2
  1. 1.Department of Applied Visual ScienceOsaka University Medical SchoolOsakaJapan
  2. 2.Department of OphthalmologyOsaka University Medical SchoolOsakaJapan
  3. 3.Department of Molecular NeurobiologyTokyo Metropolitan Institute for NeuroscienceTokyoJapan

Personalised recommendations