Formation of hippocampal synapses on patterned substrates of a laminin-derived synthetic peptide
Formation of hippocampal synapses on patterned substrates of a laminin-derived synthetic peptide
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DOI：
10.1046/j.1460-9568.2000.00977.x
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发表时间：
2000-03-01
影响因子：
3.4
通讯作者：
Kano, M
中科院分区：
文献类型：
--
作者：
Matsuzawa, M;Tabata, T;Kano, M
我们创造了一种新的培养系统，它提供了简单、刻板的神经元电路，适合于研究哺乳动物中枢神经元之间的突触事件。我们使用表面化学和激光光刻来产生神经元相容的衬底的几何图案，这些基片由较少神经元相容的表面隔开。图案化的底物由层粘连蛋白衍生的合成肽PA22-2和十烷基二甲基硅烷(DDMS)或三甲基硅烷(TMS)的间隔面组成。分离的大鼠海马神经元在没有胶质细胞帮助的情况下在有图案的底物上存活了几天，并沿着底物延伸它们的神经突起。TMS间隔面比DDMS间隔面更有利于海马神经元兴奋性发育和轴突分化，但不利于静息电导的发育。此外，生长在有图案的底物上的神经元之间经常进行突触素阳性的接触。从这些神经元记录的自发突触后电流表明，这些接触确实是功能性突触。当海马神经元以非常低的密度电镀时，它们通常在图案化的衬底上形成只由两个神经元组成的电路。这样一个简单的电路使我们能够分析单个神经元对中的突触传递，而不受第三个神经元的影响。随着分析的清晰和操作的准备，我们的培养系统将为研究哺乳动物中央突触的发育和功能提供一个强大的工具。
We created a new culture system which provides simple, stereotyped neuronal circuitries suitable for investigating synaptic events between mammalian central neurons. We used surface chemistry and laser-lithography to produce geometrical patterns of neuron-compatible substrate spaced by less neuron-compatible surfaces. The patterned substrates were composed of a laminin-derived synthetic peptide, PA22-2, and the spacing surfaces of either decyldimethylsilane (DDMS) or trimethylsilane (TMS). Dissociated rat hippocampal neurons survived on the patterned substrates for several days without the aid of glia and extended their neurites along the substrates. The TMS spacing surfaces appeared more favourable for the excitability development and axonal differentiation of the hippocampal neurons, but less favourable for the development of the resting conductance than the DDMS spacing surfaces. Furthermore, neurons grown on the patterned substrates frequently made synaptophysin-positive contacts with one another. Spontaneous post-synaptic currents recorded from such neurons suggest that these contacts were indeed functional synapses. When hippocampal neurons were plated at a very low density, they often formed circuitries consisting of only two neurons on the patterned substrate. Such a simple circuitry allowed us to analyse synaptic transmission in a single neuronal pair without the influence of the third neurons. With the clarity of analysis and the readiness of manipulation, our culture system would offer a powerful tool for studying development and functions of mammalian central synapses.