June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Melatonin protects 661W cells from cell death induced by H2O2 via inhibition of the Fas/FasL-Caspase 3 pathway
Author Affiliations & Notes
  • Aida Sanchez-Bretano
    Pharmacology, Morehouse School of Medicine, Atlanta, Georgia, United States
  • Uzair Janjua
    Pharmacology, Morehouse School of Medicine, Atlanta, Georgia, United States
  • ilaria Piano
    Dipartimento di Farmacia, Universita di Pisa, Pisa, Italy
    Pharmacology, Morehouse School of Medicine, Atlanta, Georgia, United States
  • Glaudia Gargini
    Dipartimento di Farmacia, Universita di Pisa, Pisa, Italy
  • Kenkichi Baba
    Pharmacology, Morehouse School of Medicine, Atlanta, Georgia, United States
  • Gianluca Tosini
    Pharmacology, Morehouse School of Medicine, Atlanta, Georgia, United States
  • Footnotes
    Commercial Relationships   Aida Sanchez-Bretano, None; Uzair Janjua, None; ilaria Piano, None; Glaudia Gargini, None; Kenkichi Baba, None; Gianluca Tosini, None
  • Footnotes
    Support  GM116760, EY022216, EY026291, EY020821, GM109861
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 354. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Aida Sanchez-Bretano, Uzair Janjua, ilaria Piano, Glaudia Gargini, Kenkichi Baba, Gianluca Tosini; Melatonin protects 661W cells from cell death induced by H2O2 via inhibition of the Fas/FasL-Caspase 3 pathway. Invest. Ophthalmol. Vis. Sci. 2017;58(8):354.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose : Previous studies have shown that melatonin (MEL) signaling is involved in the modulation of photoreceptor viability during aging. Recent work from our laboratory has suggested that MEL may protect cones by modulating the Fas/FasL-Caspase 3 pathway. We have also reported that 661W cells - a photoreceptor-like cell line – express MEL receptors (MT1 and MT2) mRNAs. In this study, we first investigated the signaling pathway activated by MEL in 661W cells and then whether MEL can protect these cells from H2O2 induced cell death.

Methods : MEL receptors presence in 661W was studied by immunohistochemistry (IHC). 661W were treated with MEL and MEL agonist and antagonist to identify the intracellular pathways activated by MEL in these cells. To test the protective effect, 661W cells were subjected to a 2-h treatment with H2O2 and/or MEL. To analyze the pathways involved in H2O2–mediated cell death, Fas/FasL and caspase 3 were analyzed at the level or mRNA expression and protein abundance by western blot. Cell viability was analyzed by using trypan blue.

Results : MEL receptors immunoreactivity was observed in 661W cells by IHC. The increase in cAMP production induced by forskolin was inhibited by MEL and IIK7 (a melatonin agonist) in a dose-dependent manner (p<0.05). These results suggest that in 661W cells MEL receptors are functional and - also in these cells - MT1 and MT2 receptors may heterodimerize. MEL partially prevented the H2O2-mediated cell death (20-25%; p<0.05). This effect was replicate with IIK7 when used at 1 µM (p<0.05). Pre-incubation with luzindole, a MEL antagonist, blocked MEL protection (p<0.05). Fas, FasL and caspase-3 gene expression was increased in cells treated with H2O2 and this effect was prevented by MEL (p<0.05). Finally, Kp7-6, an antagonist of Fas/FasL, blocked the cell death caused by H2O2 (p<0.05). Melatonin treatment also partially prevented the activation of Caspase 3 caused by H2O2.

Conclusions : Our results demonstrate that MEL receptors are present and functional in 661W. MEL can prevent photoreceptor cell death induced by H2O2 via the inhibition of the pro-apoptotic pathway Fas/FasL-Caspase 3. Finally, our data indicate that 661W cells may represent a new useful model to study melatonin signaling in a photoreceptor-like cell line.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×