首都医科大学学报 ›› 2009, Vol. 30 ›› Issue (5): 601-606.doi: 10.3969/j.issn.1006-7795.2009.05.007

• PD的发病机制与早期诊断 • 上一篇    下一篇

视交叉上核脑片与NIH/3T3成纤维细胞共培养模型的建立

左晓虹1, 蔡彦宁1, 李宁1,2, 于顺1, 张燕莉1, 陈彪1   

  1. 1. 首都医科大学宣武医院老年病研究所神经生物学研究室,神经变性病教育部重点实验室;2. 首都师范大学生命科学学院细胞发育研究室
  • 收稿日期:2009-07-16 修回日期:1900-01-01 出版日期:2009-10-21 发布日期:2009-10-21
  • 通讯作者: 蔡彦宁

Establishment of a Novel Co-culture Model of Suprachiasmatic Nucleus Slices and NIH/3T3 Cells in a Serum-free Condition

ZUO Xiao-hong1, CAI Yan-ning1, LI Ning1,2, YU Shun1, ZHANG Yan-li1, CHEN Biao1   

  1. 1. Department of Neurobiology, Beijing Institute of Geriatrics, Xuanwu Hospital, Capital Medical University; Key Laboratory of Neurodegenerative Diseases of Ministry of Education;2. Laboratory of Cell and Development, College of Life Sciences, Capital Normal University
  • Received:2009-07-16 Revised:1900-01-01 Online:2009-10-21 Published:2009-10-21

摘要: 目的 建立视交叉上核(suprachiasmatic nucleus,SCN)脑片与NIH/3T3成纤维细胞共培养的分子时钟诱导模型。方法 选用7日龄SD大鼠,分离SCN脑片或皮层脑片,分别与NIH/3T3细胞共培养,分为SCN共培养组和对照组皮质共培养组,6 d后连续24 h每间隔6 h收集NIH/3T3细胞,抽提细胞总RNA,实时定量RT-PCR检测时钟基因Per1mRNA水平。结果 SCN共培养组Per1的表达均表现明显的节律性(P=0.001),对照组大脑皮质共培养组Per1基因表达均无明显节律性变化。结论 成功建立SCN脑片与NIH3T3细胞共培养模型并诱导细胞内分子时钟的节律性表达,为研究SCN调控外周组织细胞生物节律提供有力工具。

关键词: 昼夜节律, 表达, 视交叉上核, Per1

Abstract: Objective To establish a novel co-cultures model of suprachiasmatic nucleus(SCN) slices and NIH/3T3 cells in a serum-free condition. Methods SCN or cortical slices prepared from 7-day-old male SD rats were cultured with NIH/3T3 cells for 6 d. 24 h expression profile of Per1 was examined in NIH/3T3 cells using real-time PCR. Results A significant daily oscillation of Per1 mRNA expression was observed in NIH/3T3 cells co-cultured with SCN slices. The peak time of Per1 was at CT11. No daily oscillation was found in NIH/3T3 cells induced with cortical slices. Conclusion A novel co-culture model of SCN slices and NIH/3T3 cells was established, which will facilitate the studies on the communications between SCN and peripheral tissues.

Key words: circadian, expression, hypothalamic suprachiasmatic nucleus, Per1

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