The hippocampal mossy fiber (MF) terminal is among the largest and most complex synaptic structures in the brain. Our understanding of the development of this morphologically elaborate structure has been limited because of the inability of standard electron microscopy techniques to quickly and accurately reconstruct large volumes of neuropil. Here we use serial block-face electron microscopy (SBEM) to surmount these limitations and investigate the establishment of MF connectivity during mouse postnatal development. Based on volume reconstructions, we find that MF axons initially form bouton-like specializations directly onto dendritic shafts, that dendritic protrusions primarily arise independently of bouton contact sites, and that a dramatic increase in presynaptic and postsynaptic complexity follows the association of MF boutons with CA3 dendritic protrusions. We also identify a transient period of MF bouton filopodial exploration, followed by refinement of sites of synaptic connectivity. These observations enhance our understanding of the development of this highly specialized synapse and illustrate the power of SBEM to resolve details of developing microcircuits at a level not easily attainable with conventional approaches.

译文

海马苔藓纤维 (MF) 末端是大脑中最大,最复杂的突触结构之一。由于标准电子显微镜技术无法快速准确地重建大量神经纤维,因此我们对这种形态精细结构的发展的理解受到限制。在这里,我们使用连续块面电子显微镜 (SBEM) 克服这些限制,并研究小鼠出生后发育过程中MF连接的建立。基于体积重建,我们发现MF轴突最初直接在树枝状轴上形成类似bouton的特化,树枝状突起主要独立于bouton接触部位产生,并且突触前和突触后复杂性的急剧增加伴随着MF bouton与CA3树枝状突起的关联。我们还确定了MF bouton丝状探索的瞬态,然后完善了突触连接位点。这些观察结果增强了我们对这种高度专业化的突触发展的理解,并说明了SBEM在传统方法难以达到的水平上解决微电路开发细节的能力。

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