September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Circadian Rhythm in Cone-like (661W) Photoreceptor Cells
Author Affiliations & Notes
  • Kenkichi Baba
    Pharmacology/Toxicology, Morehouse School of Medicine, Atlanta, Georgia, United States
  • Takana Tubo
    Pharmacology/Toxicology, Morehouse School of Medicine, Atlanta, Georgia, United States
  • Ting-Chung Suen
    Pharmacology/Toxicology, Morehouse School of Medicine, Atlanta, Georgia, United States
  • Jason DeBruyne
    Pharmacology/Toxicology, Morehouse School of Medicine, Atlanta, Georgia, United States
  • Gianluca Tosini
    Pharmacology/Toxicology, Morehouse School of Medicine, Atlanta, Georgia, United States
  • Footnotes
    Commercial Relationships   Kenkichi Baba, None; Takana Tubo, None; Ting-Chung Suen, None; Jason DeBruyne, None; Gianluca Tosini, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 4663. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to Subscribers Only
      Sign In or Create an Account ×
    • Get Citation

      Kenkichi Baba, Takana Tubo, Ting-Chung Suen, Jason DeBruyne, Gianluca Tosini; Circadian Rhythm in Cone-like (661W) Photoreceptor Cells. Invest. Ophthalmol. Vis. Sci. 2016;57(12):4663.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : Previous studies have shown that the mammalian retina contains circadian clocks that control many physiological events with this tissue. However, the presence of circadian clocks within the photoreceptor (rods or cones) is still matter of debate. In this study, we investigate whether cone-like cells (661W, a murine photoreceptor derived cell line) contain circadian clocks and whether they express mRNA for dopamine and melatonin receptors.

Methods : 661W cells generously provided by Dr. Al-Ubaidi, University of Oklahoma Health Science Center were used in this study. Cells were cultured and grown to 90-100 % confluency with Dulbecco’s Modified Eagle Medium (DMEM) at 37 oC and then were harvested and the mRNA was extracted. The presence of mRNAs for clock genes (Period1, Period2 and Bmal1), dopamine (D1-D5) and melatonin (MT1 and MT2) receptors was investigated with RT-PCR. To determine the presence of a circadian clock, 661W cells were stably transfected with a PERIOD2::LUCIFERASE (PER2::LUC) . Transfected cells (661W-PER2::LUC) were then cultured with DMEM containing 0.1mM D-Luciferin K salt, and the bioluminescence rhythms emitted from 661W-PER2::LUC cells was measured using a LumiCycle®.

Results : RT-PCR amplification and electrophoresis gel analysis indicated that Period1, Period2, Bmal1 mRNAs were all present in 661W cells. mRNAs for the Dopamine D4 and both melatonin (MT1 and MT2) receptors was also amplifiable from these cells. 661W-PER2::LUC cells also expressed a clear circadian rhythm in bioluminescence that lasted for at least 4 days The average free running period was 21.96±0.26 hours (Mean±SEM, n=16).

Conclusions : Our data indicate that 661W-PER2::LUC cells contain a circadian clock and express dopamine and melatonin receptors. Thus 661W-PER2::LUC cells may represent an useful in vitro model to study the mechanisms that generate circadian rhythms and the role that melatonin and dopamine plays in the entrainment of circadian rhythm in a cone-like photoreceptor.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

×
×

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.

×