BACKGROUND:Spinal motor nerves are essential for relaying information between the central and peripheral nervous systems. Perturbations to cell types that comprise these nerves may impair rapid and efficient transmission of action potentials and alter nerve function. Identifying ultrastructural changes resulting from defects to these cellular components via transmission electron microscopy (TEM) can provide valuable insight into nerve function and disease. However, efficiently locating spinal motor nerves in adult zebrafish for TEM is challenging and time-consuming. Because of this, we developed a protocol that allows us to quickly and precisely locate spinal motor nerve roots in adult zebrafish for TEM processing. RESULTS:Following fixation, a transverse slab of adult zebrafish dissected from the trunk region was mounted in embedding media, sectioned, and secondary fixation with osmium tetroxide performed. Transverse sections containing motor nerves were selected for TEM ultrathin sectioning and imaging. CONCLUSIONS:We developed an efficient protocol for locating spinal motor nerves in adult zebrafish to allow for ultrastructural characterization. Although our work focuses on spinal motor nerves, this protocol may be useful for efficiently identifying other discrete, repeated structures within the developing and mature nervous system that are difficult to find via traditional, whole organism TEM processing. Developmental Dynamics 246:956-962, 2017. © 2017 Wiley Periodicals, Inc.

译文

背景:脊髓运动神经对于在中枢神经系统和周围神经系统之间传递信息至关重要。对包含这些神经的细胞类型的干扰可能会损害动作电位的快速有效传递并改变神经功能。通过透射电子显微镜(TEM)识别出这些细胞成分的缺陷所导致的超微结构变化,可以为神经功能和疾病的诊断提供有价值的信息。然而,有效地定位成年斑马鱼中的脊髓运动神经以进行TEM既具有挑战性又耗时。因此,我们开发了一种协议,该协议使我们能够快速而精确地定位成年斑马鱼中的脊髓运动神经根,以进行TEM处理。
结果:固定后,将成年斑马鱼从躯干区域切下的横板固定在包埋介质中,切片,并用四氧化进行二次固定。选择含有运动神经的横断面进行TEM超薄切片和成像。
结论:我们开发了一种有效的协议,用于定位成年斑马鱼的脊髓运动神经,以便进行超微结构表征。尽管我们的工作集中在脊髓运动神经上,但该协议可能对有效识别发育中和成熟的神经系统中其他离散的,重复的结构非常有用,而这些结构很难通过传统的全机体TEM处理找到。 Developmental Dynamics 246:956-962,2017.©2017 Wiley Periodicals,Inc.

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