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The Nanoscale Observation of the Three-Dimensional Structures of Neurosynapses, Membranous Conjunctions Between Cultured Hippocampal Neurons and Their Significance in the Development of Epilepsy

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Abstract

The nanoscale three-dimensional structures of neurosynapses are unknown, and the neuroanatomical basis of epilepsy remains to be elucidated. Here, we studied the nanoscale three-dimensional synapses between hippocampal neurons, and membranous conjunctions between neurons were found with atomic force microscopy (AFM) and confirmed by transmission electron microscope (TEM), and their pathophysiological significance was primarily investigated. The neurons and dendrites were marked by MAP-2, axons by neurofilament 200, and synapses by synapsin I immunological staining. In the synapsin I-positive neurite ends of the neurons positively stained with MAP-2 and neurofilament 200, neurosynapses with various nanoscale morphology and structure could be found by AFM. The neurosynapses had typical three-dimensional structures of synaptic triplet including the presynaptic neurite end, synaptic cleft of 30 ∼ 40 in chemical synapses and 2 ∼ 6 nm in electrical ones, the postsynaptic neurite or dendrite spine, the typical neurite end button, the distinct pre- and postsynaptic membranes, and the obvious thickening of the postsynaptic membranes or neurites. Some membranous connections including membrane-like junctions (MLJ) and fiber-tube links (FTL) without triplet structures and cleft were found between neurons. The development frequencies of the two membranous conjunctions increased while those of the synaptic conjunctions decreased between the neurons from Otx1 knock-out mice in comparison with those between the neurons from normal mice. These results suggested that the neuroanatomical basis of Otx1 knock-out epilepsy is the combination of the decreased synaptic conjunctions and the increased membranous conjunctions.

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Acknowledgments

This work was supported by the Major Special Projection for Nanoscience of China (2010CB933904) and the Major Special Projection for New Drugs of China (2011ZX09102-001-15).

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Authors and Affiliations

  1. Key Laboratory of Nanopharmacology and Nanotoxicology, Beijing Institute of Pharmacology and Toxicology, 27 Taiping Road, Beijing, 100850, China

    Lan Sun, Shuang Jiang, Xianhua Tang, Yingge Zhang & Xia Jiang

  2. Neurosciences Research Institute, Peking University, Key Laboratory of Neuroscience; Department of Neurobiology, Peking University Health Science Center, 38 Xue Yuan Road, Beijing, 100083, China

    Luye Qin & Albert Cheung Hoi Yu

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  1. Lan Sun
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  2. Shuang Jiang
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Sun, L., Jiang, S., Tang, X. et al. The Nanoscale Observation of the Three-Dimensional Structures of Neurosynapses, Membranous Conjunctions Between Cultured Hippocampal Neurons and Their Significance in the Development of Epilepsy. Mol Neurobiol 53, 7137–7157 (2016). https://doi.org/10.1007/s12035-015-9588-1

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