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Related Experiment Videos

HEK293S cells have functional retinoid processing machinery.

Lioubov I Brueggemann1, Jack M Sullivan

  • 1Department of Ophthalmology, Institute for Human Performance, State University of New York, Upstate Medical University, 750 East Adams Street, Syracuse, NY 13210, USA.

The Journal of General Physiology
|May 30, 2002
PubMed
Summary
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Human embryonic kidney cells expressing rhodopsin can regenerate visual pigment through intrinsic retinoid processing. This study reveals HEK293S cells possess machinery for converting Vitamin A and all-trans-retinal into the active form, enabling early receptor current signal recovery.

Area of Science:

  • Biochemistry
  • Cell Biology
  • Neuroscience

Background:

  • Rhodopsin activation, crucial for vision, is measured by the early receptor current (ERC), a charge motion.
  • ERC signals in opsin-expressing cells recover after rhodopsin bleaching through dark adaptation.
  • The source of this ERC signal recovery in human embryonic kidney (HEK293S) cells remains to be fully elucidated.

Purpose of the Study:

  • To investigate the intrinsic mechanisms responsible for early receptor current (ERC) signal recovery in HEK293S cells.
  • To determine if HEK293S cells can regenerate ground-state rhodopsin from various retinoid precursors.
  • To assess the role of the retinoid cycle in this regeneration process.

Main Methods:

  • HEK293S cells expressing wild-type human rod opsin were loaded with 11-cis-retinal, all-trans-retinal, or Vitamin A.

Related Experiment Videos

  • Early receptor currents (ERCs) were elicited by flash photolysis and measured using whole-cell recording.
  • The effect of 4-butylaniline, a retinoid cycle inhibitor, on ERC recovery was evaluated.

    • Main Results:

    • Cells loaded with 11-cis-retinal showed bimodal ERC signals (R(1), R(2)) after regeneration.
    • Cells regenerated with all-trans-retinal or Vitamin A exhibited negative or absent ERCs initially.
    • Overnight dark adaptation led to recovery of outward R(2) signals in cells treated with all-trans-retinal or Vitamin A, indicating retinoid conversion.
    • 4-butylaniline treatment reversibly suppressed the recovery of the R(2) component.

    Conclusions:

    • HEK293S cells possess intrinsic retinoid processing capabilities, converting all-trans-retinal and Vitamin A into cis-retinaldehyde.
    • This endogenous machinery regenerates ground-state rhodopsin, leading to the recovery of ERC signals.
    • These findings suggest a functional similarity between retinoid processing in HEK293S cells and the mammalian eye.