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Name pinyin
LIANG Fanghao
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0710 生物学
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07 理学
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如兴奋性三突触回路( EC→DG→CA3→CA1→EC) 等,非常有利于神经元之间
的信号调控研究。 在神经元的信号调控中, 过去一般认为是单向的,即由突触
前神经元通过突触释放化学或电信号到突触后神经元并调控其功能。 但是近年
来越来越多的证据表明,神经元可以通过突触实现双向传递信息。 本课题希望
建立一套基于微流控系统的海马体神经元体外突触体系,用于研究海马体中 CA
锥体神经元(突触后) 释放的信号通过突触来逆向调控齿状回( DG)颗粒细胞
(突触前) 的基因表达和发育。主要研究内容包括探索高效分离海马体不同神
经元的方法, 设计与组装微流控系统, 验证神经元体外培养中的突触形成, FAST
DiI 染料逆向标记的神经元的流式分选, RNA 提取以用于 RNA 测序和转录组
学分析等。其中主要用到大脑精细结构解剖技术、 神经元体外培养技术、 细胞
荧光流式分选技术、逆向荧光染料标记技术和 RNA 提取等研究方法。本课题
流体分隔培养, 同时可以使神经元之间建立突触联系, 于是拥有可以对一种或
体神经元体外微流控培养体系, 不仅能实现逆向信号的调控研究,还可以方便

Other Abstract

The hippocampus is one of the most popular brain regions studied by
neuroscientists so far. Its unique structural features, such as excitatory three-synaptic
circuits (EC→DG→CA3→CA1→EC), are very useful for signal regulation studies
between neurons. In the signal regulation of neurons, it is generally considered to be
unidirectional, that is, the presynaptic neurons release chemical and electrical signals
to the postsynaptic neurons via synapses and regulate it. But in recent years, there is
increasing evidence that neurons can transmit information bidirectionally through
synapses. This project aims to establish an in vitro synapse formation system of
hippocampal neurons based on microfluidic devices for studying retrograde
regulation of gene expression and development of dentate gyrus (DG) granule cells
(presynaptic) by signal(s) released by CA pyramidal neurons (postsynaptic) in a
retrograde manner. The main research contents include exploring methods for
efficiently dissecting and separating different neurons of hippocampus, designing and
assembling microfluidic systems, verifying synapse formation in vitro cell culture,
sorting specific cells of FAST DiI retrograde labeling, RNA extraction to prepare for
RNA sequencing and transcriptomic analysis. The techniques and methods used in
this study are fine brain structure anatomy, FACS sorting, in vitro neuron culture,
retrograde labeling by fluorochrome and RNA extraction. The innovation of this
subject lies in the combination of microfluidic technology and neuronal culture, so
that a variety of neurons can be cultured separately under the same conditions, and
at the same time, synaptic connections can be established between the neurons. This
opens up the possibility of processing one or more neurons conveniently so that it is
very easy to collect the specific neurons after the retrograde signal regulation and
conduct the later research. The main significance of this topic is firstly to establish a
set of microfluidic cell culture system of hippocampal neurons in vitro, which can
not only realize the study of retrograde signal regulation, but also can be easily
applied to other signal regulation and other neuronal cultures; secondly, since
progress in retrograde signal regulation is limited, we hope this project can open a
new path in this field.

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梁芳昊. 小鼠海马神经元发育的信号调控研究[D]. 哈尔滨. 哈尔滨工业大学,2019.
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