J Korean Neuropsychiatr Assoc.
2006 Jan;45(1):21-27.
Regulation of the Circadian Gene CLOCK Expression by KCl Depolarization in B35 Rat Neuroblastoma Cells
- Affiliations
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- 1Clinical Research Institute, Seoul National University Hospital, Seoul, Korea.
- 2Department of Psychiatry, Seoul National University Hospital, Seoul, Korea. aym@snu.ac.kr
- 3Department of Psychiatry and Institute of Human Behavioral Medicine, Seoul National University College of Medicine, Seoul, Korea.
Abstract
OBJECTIVES
To investigate the effects of KCl on regulation of circadian gene CLOCK expression, we observed whether induction of CLOCK is influenced by KCl depolarization in B35 rat neuroblastoma cells.
METHODS
B35 rat neuroblastoma cells were grown in Dulbecco's modified Eagle's medium supplemented with 10% FBS and 1% penicillin-streptomycin in a 37 degrees C humidified incubator with 5% CO2. Inhibitors including cycloheximide and actinomycin D were pretreated 1 hour before treatment with 50mM KCl. Immunoblotting with anti-CLOCK antibody was done.
RESULTS
CLCOK is induced by 50 mM KCl in B35 Rat Neuroblastoma cells, and a maximal induction in CLOCK level reached peak at 8 to 20 hours. The pretreatment of cycloheximide and actinomycin D prevented the induction of CLOCK by 50 mM KCl.
CONCLUSION
We suggest that KCl depolarization may play critical roles in several aspects of the circadian gene CLOCK expression.
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