Kruss Sebastian, Salem Daniel P, Vuković Lela, Lima Barbara, Vander Ende Emma, Boyden Edward S, Strano Michael S
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139.
Department of Physics, University of Illinois at Urbana-Champaign, Champaign, IL 61801.
Proc Natl Acad Sci U S A. 2017 Feb 21;114(8):1789-1794. doi: 10.1073/pnas.1613541114. Epub 2017 Feb 8.
Intercellular communication via chemical signaling proceeds with both spatial and temporal components, but analytical tools, such as microfabricated electrodes, have been limited to just a few probes per cell. In this work, we use a nonphotobleaching fluorescent nanosensor array based on single-walled carbon nanotubes (SWCNTs) rendered selective to dopamine to study its release from PC12 neuroprogenitor cells at a resolution exceeding 20,000 sensors per cell. This allows the spatial and temporal dynamics of dopamine release, following K stimulation, to be measured at exceedingly high resolution. We observe localized, unlabeled release sites of dopamine spanning 100 ms to seconds that correlate with protrusions but not predominately the positive curvature associated with the tips of cellular protrusions as intuitively expected. The results illustrate how directionality of chemical signaling is shaped by membrane morphology, and highlight the advantages of nanosensor arrays that can provide high spatial and temporal resolution of chemical signaling.
通过化学信号进行的细胞间通讯具有空间和时间两个维度,但诸如微加工电极等分析工具,每个细胞只能使用少数几个探针。在这项工作中,我们使用了一种基于对多巴胺具有选择性的单壁碳纳米管(SWCNT)的非光漂白荧光纳米传感器阵列,以超过每个细胞20,000个传感器的分辨率研究其从PC12神经祖细胞中的释放。这使得在钾离子刺激后,能够以极高的分辨率测量多巴胺释放的空间和时间动态。我们观察到多巴胺的局部、未标记的释放位点,其持续时间从100毫秒到数秒不等,这些位点与细胞突起相关,但并不主要与细胞突起尖端的正曲率相关,而这是直观上所预期的。结果表明了化学信号的方向性是如何由膜形态塑造的,并突出了纳米传感器阵列在提供化学信号高空间和时间分辨率方面的优势。