小鼠脑中蓝斑核的定位
Localization of the Locus Coeruleus in the Mouse Brain.
作者信息
Schmidt Katharina, Bari Bilal, Ralle Martina, Washington-Hughes Clorissa, Muchenditsi Abigael, Maxey Evan, Lutsenko Svetlana
机构信息
Department of Physiology, Johns Hopkins University, School of Medicine;
Department of Neuroscience, Johns Hopkins University.
出版信息
J Vis Exp. 2019 Mar 7(145). doi: 10.3791/58652.
The locus coeruleus (LC) is a major hub of norepinephrine producing neurons that modulate a number of physiological functions. Structural or functional abnormalities of LC impact several brain regions including cortex, hippocampus, and cerebellum and may contribute to depression, bipolar disorder, anxiety, as well as Parkinson disease and Alzheimer disease. These disorders are often associated with metal misbalance, but the role of metals in LC is only partially understood. Morphologic and functional studies of LC are needed to better understand the human pathologies and contribution of metals. Mice are a widely used experimental model, but the mouse LC is small (~0.3 mm diameter) and hard to identify for a non-expert. Here, we describe a step-by-step immunohistochemistry-based protocol to localize the LC in the mouse brain. Dopamine-β-hydroxylase (DBH), and alternatively, tyrosine hydroxylase (TH), both enzymes highly expressed in the LC, are used as immunohistochemical markers in brain slices. Sections adjacent to LC-containing sections can be used for further analysis, including histology for morphological studies, metabolic testing, as well as metal imaging by X-ray fluorescence microscopy (XFM).
蓝斑（LC）是去甲肾上腺素能神经元的主要枢纽，可调节多种生理功能。LC的结构或功能异常会影响包括皮层、海马体和小脑在内的多个脑区，并可能导致抑郁症、双相情感障碍、焦虑症以及帕金森病和阿尔茨海默病。这些疾病通常与金属失衡有关，但金属在LC中的作用仅得到部分了解。需要对LC进行形态学和功能研究，以更好地理解人类病理学以及金属的作用。小鼠是一种广泛使用的实验模型，但小鼠的LC很小（直径约0.3毫米），对于非专业人员来说很难识别。在这里，我们描述了一种基于免疫组织化学的分步方案，用于在小鼠脑中定位LC。多巴胺-β-羟化酶（DBH），或者酪氨酸羟化酶（TH），这两种在LC中高度表达的酶，被用作脑切片的免疫组织化学标记物。与含LC切片相邻的切片可用于进一步分析，包括用于形态学研究的组织学、代谢测试以及通过X射线荧光显微镜（XFM）进行的金属成像。
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