摘要
目的研究神经黏附分子Necl1在神经突触形成过程中的功能。方法采用半定量逆转录-聚合酶链反应(RT-PCR)方法检测在体外神经分化细胞模型(SH-SY5Y,P19)中Necl1表达量的变化;Western blot方法检测原代培养的大鼠海马和皮层神经元在体外培养条件下Necl1的表达量变化和在突触小体中Necl1的表达;细胞免疫荧光法检测神经元和293细胞共培养后293细胞表面突触形成情况,并检测在神经元内过表达Necl1后神经元表面突触密度变化。结果Necl1的表达量可随神经细胞的分化或原代神经元体外培养时间的延长而升高。纯化后的突触小体中存在Necl1的表达。过表达Necl1的293细胞在与原代神经元共培养的情况下可产生突触样结构。直接在原代培养神经元内过表达Necl1可提高神经元间突触密度。结论Necl1可能在中枢神经系统的神经突触形成中发挥功能。
Objective To study the role of cell adhesion molecules Necll in synaptogenesis in primary cultured neurons. Methods Semi-quantitive reverse transcription polymerase chain reaction （RT-PCR） was used to detect the expression pattern of Necll in the neuronal differentiation cell model in vitro. Western blot was performed to detect the expression pattern of Necll in primary cultured rat neurons and in purified synaptosome. Immunofluoresence was used to detect the synapse formation in primary neurons and in 293 cells coculture and to detect the density of synapses in primary neuron with ectopic expression of Necll. Results Necll expression increased after retinoic acid （RA） induction in SH-SY5Y and P19 cells. The increase of Necll expression was consistent with the days of primary neurons culture in vitro, and Necll partly localized in synaptosome. The overexpression of Necll in 293 ceils induced the synapse formation between cocultured 293 cells and neurons. Ectopic expression of Necll in primary neurons increased the density of synapses. Conclusion Necll plays an important role in neuronal synapse formation.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2008年第3期275-279,I0001,共6页
Acta Academiae Medicinae Sinicae
基金
国家自然科学基金(30571039、30430200)
国家高技术研究发展计划项目(863计划)(2006AA02Z137)
国家重点基础研究发展计划项目(973计划)(2005CB522507)~~
