Kosar T Fettah, Tourovskaia Anna, Figueroa-Masot Xavier, Adams Marvin E, Folch Albert
Department of Bioengineering, University of Washington, Campus Box 352255, Seattle, WA 98195-2255, USA.
Lab Chip. 2006 May;6(5):632-8. doi: 10.1039/b517475a. Epub 2006 Mar 22.
The release of synaptogenic factors by the nerve terminal plays a central role in the aggregation of neurotransmitter receptors at the postsynaptic membrane, a precisely timed and localized process that is essential for the correct formation and functioning of the synapse. This process has been difficult to re-capitulate in cell culture because present cell stimulation methods do not have sufficient spatiotemporal control of the delivery of soluble signals. We cultured myotubes atop nanofabricated planar apertures (2-8 microm diameter) to focally stimulate the muscle cell membrane with neural agrin, a synaptogenic factor released by motor neurons during development. Focal agrin delivery through the apertures after myotube fusion results in local aggregation of acetylcholine receptors (AChRs) in the vicinity of the apertures, a process reminiscent of AChR clustering at innervation sites. Since the apertures are spatially organized in microarrays, multiple experiments can be run in parallel on one device. The technique has wide applicability in cell-cell communication studies and cell-based bioassays.
神经末梢释放的促突触形成因子在神经递质受体于突触后膜聚集过程中起核心作用，这是一个时机精确且定位明确的过程，对突触的正确形成和功能至关重要。该过程在细胞培养中难以重现，因为目前的细胞刺激方法对可溶性信号传递缺乏足够的时空控制。我们将肌管培养在纳米制造的平面小孔（直径2 - 8微米）上，用神经聚集蛋白局部刺激肌细胞膜，神经聚集蛋白是运动神经元在发育过程中释放的一种促突触形成因子。肌管融合后通过小孔局部递送神经聚集蛋白会导致小孔附近乙酰胆碱受体（AChRs）局部聚集，这一过程让人联想到神经支配部位的AChR簇集。由于小孔在微阵列中呈空间排列，因此可以在一个装置上并行进行多个实验。该技术在细胞间通讯研究和基于细胞的生物测定中具有广泛的适用性。
