Regulation of the Circadian Gene CLOCK Expression by KCl Depolarization in B35 Rat Neuroblastoma Cells / 신경정신의학

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.