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成年转基因斑马鱼脊髓中的神经元标记模式。

Neuronal labeling patterns in the spinal cord of adult transgenic Zebrafish.

作者信息

Stil Aurélie, Drapeau Pierre

机构信息

Hospital Research Centre (CRCHUM) and Department of Neurosciences, Université de Montréal, Montréal, Quebec, Canada, H2X 0A9.

出版信息

Dev Neurobiol. 2016 Jun;76(6):642-60. doi: 10.1002/dneu.22350. Epub 2015 Sep 26.

Abstract

We describe neuronal patterns in the spinal cord of adult zebrafish. We studied the distribution of cells and processes in the three spinal regions reported in the literature: the 8th vertebra used as a transection injury site, the 15th vertebra mainly used for motor cell recordings and also for crush injury, and the 24th vertebra used to record motor nerve activity. We used well-known transgenic lines in which expression of green fluorescent protein (GFP) is driven by promoters to hb9 and isl1 in motoneurons, alx/chx10 and evx1 interneurons, ngn1 in sensory neurons and olig2 in oligodendrocytes, as well as antibodies for neurons (HuC/D, NF and SV2) and glia (GFAP). In isl1:GFP fish, GFP-positive processes are retained in the upper part of ventral horns and two subsets of cell bodies are observed. The pattern of the transgene in hb9:GFP adults is more diffuse and fibers are present broadly through the adult spinal cord. In alx/chx10 and evx1 lines we respectively observed two and three different GFP-positive populations. Finally, the ngn1:GFP transgene identifies dorsal root ganglion and some cells in dorsal horns. Interestingly some GFP positive fibers in ngn1:GFP fish are located around Mauthner axons and their density seems to be related to a rostrocaudal gradient. Many other cell types have been described in embryos and need to be studied in adults. Our findings provide a reference for further studies on spinal cytoarchitecture. Combined with physiological, histological and pathological/traumatic approaches, these studies will help clarify the operation of spinal locomotor circuits of adult zebrafish.

摘要

我们描述了成年斑马鱼脊髓中的神经元模式。我们研究了文献中报道的三个脊髓区域中细胞和突起的分布:第8节椎骨用作横断损伤部位,第15节椎骨主要用于运动细胞记录以及挤压损伤,第24节椎骨用于记录运动神经活动。我们使用了著名的转基因品系,其中绿色荧光蛋白(GFP)的表达由启动子驱动,在运动神经元中由hb9和isl1驱动,在中间神经元中由alx/chx10和evx1驱动,在感觉神经元中由ngn1驱动,在少突胶质细胞中由olig2驱动,以及用于神经元(HuC/D、NF和SV2)和神经胶质(GFAP)的抗体。在isl1:GFP鱼中,GFP阳性突起保留在腹角上部,并观察到两个细胞体亚群。hb9:GFP成年鱼中转基因的模式更弥散,纤维广泛存在于成年脊髓中。在alx/chx10和evx1品系中,我们分别观察到两个和三个不同的GFP阳性群体。最后,ngn1:GFP转基因识别背根神经节和背角中的一些细胞。有趣的是,ngn1:GFP鱼中的一些GFP阳性纤维位于Mauthner轴突周围,其密度似乎与前后梯度有关。在胚胎中已经描述了许多其他细胞类型,需要在成体中进行研究。我们的发现为进一步研究脊髓细胞结构提供了参考。结合生理学、组织学和病理/创伤学方法,这些研究将有助于阐明成年斑马鱼脊髓运动回路的运作。

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