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

Jpn J Ophthalmol. 2008 May-Jun;52(3):217-223. doi: 10.1007/s10384-008-0533-y. Epub 2008 Jul 27.

Abstract

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

Methods: 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 ([Ca(2+)]) induced by ES was monitored by Ca(2+) imaging with Fura 2-AM. Ca(2+) imaging and RT-PCR were performed with and without the application of l muM nifedipine, an L-type calcium channel blocker.

Results: The mRNA level of IGF-1 was increased significantly (P<0.05) by about 1.3-fold immediately after 10 mA ES. [Ca(2+)] 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 Ca(2+) influx were suppressed by nifedipine.

Conclusions: These results indicate that the enhancement of IGF-1 transcription by ES in cultured Müller cells depends largely on Ca(2+) influx through L-type Ca(2+) channels.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / metabolism
  • Calcium Channel Blockers / pharmacology
  • Calcium Channels, L-Type / metabolism*
  • Cells, Cultured
  • Electric Stimulation*
  • Enzyme-Linked Immunosorbent Assay
  • Gene Expression Regulation / physiology*
  • Insulin-Like Growth Factor I / genetics*
  • Microscopy, Fluorescence
  • Neuroglia / metabolism*
  • Nifedipine / pharmacology
  • RNA, Messenger / metabolism
  • Rats
  • Rats, Long-Evans
  • Retina / cytology*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Up-Regulation

Substances

  • Calcium Channel Blockers
  • Calcium Channels, L-Type
  • RNA, Messenger
  • Insulin-Like Growth Factor I
  • Nifedipine
  • Calcium