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Journal of Neuroscience, Vol 8, 22-30, Copyright © 1988 by Society for Neuroscience
Circadian clock in cell culture: II. In vitro photic entrainment of melatonin oscillation from dissociated chick pineal cells
LM Robertson and JS Takahashi
Department of Neurobiology and Physiology, Northwestern University, Evanston, Illinois 60201.
The avian pineal gland contains circadian oscillators that regulate the
rhythmic synthesis of melatonin. We have developed a flow-through cell
culture system in order to begin to study the cellular and molecular basis
of this vertebrate circadian oscillator. Pineal cell cultures express a
circadian oscillation of melatonin release for at least 5 cycles in
constant darkness with a period close to 24 hr. In all circadian systems,
light regulates the rhythm by the process of entrainment that involves
control of the phase and period of the circadian oscillator. In chick
pineal cell cultures we have investigated the entraining effects of light
in 2 ways: by shifting the light-dark cycle in vitro and by measuring the
phase-shifting effects of single light pulses. A 6 hr advance or delay of a
LD 12:12 light- dark cycle produced a corresponding shift in the melatonin
rhythm. The phase shifts of the rhythms persisted after transfer to
constant darkness, showing that the underlying circadian oscillator was
entrained. Photic entrainment of the oscillator was further characterized
by measuring the phase-shifting effects of single 6 hr light pulses. Single
pulses of light shifted the phase of the circadian oscillator in a
phase-dependent manner. Light pulses beginning early in the subjective
night delayed the phase of the oscillation 8 hr relative to dark controls.
Conversely, light pulses beginning late in the subjective night advanced
the phase of the oscillation nearly 8 hr. Thus, photoreceptors within the
cell cultures can mediate entrainment of the pineal oscillators.(ABSTRACT
TRUNCATED AT 250 WORDS)
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