Sakata Yoko, Olson Jared K, Michel William C
University of Utah School of Medicine, Department of Physiology, 410 Chipeta Way Room 155, Salt Lake City, UT 84108-1297, USA.
Methods Cell Sci. 2003;25(1-2):39-48. doi: 10.1023/B:MICS.0000006852.16798.34.
Olfactory coding at the level of the olfactory bulb is thought to depend upon an ensemble response of mitral cells receiving input from chemotopically-organized projections of olfactory sensory neurons and regulated by lateral inhibitory circuits. Immunocytochemical methods are described to metabolically classify neurons in the developing zebrafish olfactory system based on the relative concentrations of taurine, glutamate, GABA (and potentially other small biogenic amines) and a small guanidium-based cation, agmatine, which labels NMDA-sensitive cells by permeating through active ionotropic glutamate receptor channels. Using metabolic profiling in conjunction with activity dependent labeling we demonstrate that neuronal differentiation in the developing olfactory bulb, as assessed by acquisition of a mature neurochemical profile, and sensitivity to an ionotropic glutamate receptor agonist, NMDA, occurs during the second day of development. This experimental approach is likely to be useful in studies concerned with the development of glutamatergic signaling pathways.
嗅球水平的嗅觉编码被认为依赖于接受嗅觉感觉神经元的化学拓扑组织投射输入并受侧向抑制回路调节的二尖瓣细胞的整体反应。本文描述了免疫细胞化学方法,用于根据牛磺酸、谷氨酸、GABA(以及可能的其他小生物胺)和一种基于胍的小阳离子胍丁胺的相对浓度,对发育中的斑马鱼嗅觉系统中的神经元进行代谢分类,胍丁胺通过活性离子型谷氨酸受体通道渗透来标记对NMDA敏感的细胞。通过将代谢谱分析与活性依赖性标记相结合,我们证明,通过获得成熟的神经化学谱评估的发育中的嗅球中的神经元分化以及对离子型谷氨酸受体激动剂NMDA的敏感性,发生在发育的第二天。这种实验方法可能在与谷氨酸能信号通路发育相关的研究中有用。