B35 백서 신경모세포종 세포에서 KCl에 의한 탈분극이 일중 주기 관련 분자 CLOCK의 발현에 미치는 영향

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

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℃ 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.