An autonomous circadian clock in the inner mouse retina regulated by dopamine and GABA

Guo Xiang Ruan, Gregg C. Allen, Shin Yamazaki, Douglas G. McMahon

Research output: Contribution to journalArticle

94 Citations (Scopus)

Abstract

The influence of the mammalian retinal circadian clock on retinal physiology and function is widely recognized, yet the cellular elements and neural regulation of retinal circadian pacemaking remain unclear due to the challenge of long-term culture of adult mammalian retina and the lack of an ideal experimental measure of the retinal circadian clock. In the current study, we developed a protocol for long-term culture of intact mouse retinas, which allows retinal circadian rhythms to be monitored in real time as luminescence rhythms from a PERIOD2::LUCIFERASE (PER2::LUC) clock gene reporter. With this in vitro assay, we studied the characteristics and location within the retina of circadian PER2::LUC rhythms, the influence of major retinal neurotransmitters, and the resetting of the retinal circadian clock by light. Retinal PER2::LUC rhythms were routinely measured from whole-mount retinal explants for 10 d and for up to 30 d. Imaging of vertical retinal slices demonstrated that the rhythmic luminescence signals were concentrated in the inner nuclear layer. Interruption of cell communication via the major neurotransmitter systems of photoreceptors and ganglion cells (melatonin and glutamate) and the inner nuclear layer (dopamine, acetylcholine, GABA, glycine, and glutamate) did not disrupt generation of retinal circadian PER2::LUC rhythms, nor did interruption of intercellular communication through sodium-dependent action potentials or connexin 36 (cx36)-containing gap junctions, indicating that PER2::LUC rhythms generation in the inner nuclear layer is likely cell autonomous. However, dopamine, acting through D1 receptors, and GABA, acting through membrane hyperpolarization and casein kinase, set the phase and amplitude of retinal PER2::LUC rhythms, respectively. Light pulses reset the phase of the in vitro retinal oscillator and dopamine D1 receptor antagonists attenuated these phase shifts. Thus, dopamine and GABA act at the molecular level of PER proteins to play key roles in the organization of the retinal circadian clock.

Original languageEnglish (US)
Article numbere249
Pages (from-to)2248-2265
Number of pages18
JournalPLoS Biology
Volume6
Issue number10
DOIs
StatePublished - Oct 2008

Fingerprint

Circadian Clocks
gamma-aminobutyric acid
luciferase
dopamine
Luciferases
retina
circadian rhythm
gamma-Aminobutyric Acid
Retina
Clocks
Dopamine
mice
cell communication
luminescence
neurotransmitters
Luminescence
glutamates
Neurotransmitter Agents
Glutamic Acid
Casein Kinases

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)

Cite this

An autonomous circadian clock in the inner mouse retina regulated by dopamine and GABA. / Ruan, Guo Xiang; Allen, Gregg C.; Yamazaki, Shin; McMahon, Douglas G.

In: PLoS Biology, Vol. 6, No. 10, e249, 10.2008, p. 2248-2265.

Research output: Contribution to journalArticle

Ruan, Guo Xiang ; Allen, Gregg C. ; Yamazaki, Shin ; McMahon, Douglas G. / An autonomous circadian clock in the inner mouse retina regulated by dopamine and GABA. In: PLoS Biology. 2008 ; Vol. 6, No. 10. pp. 2248-2265.
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