Hellmer Chase B, Ichinose Tomomi
Departments of Anatomy and Cell Biology, Wayne State University School of Medicine, Detroit, MI, USA.
Department of Ophthalmology, Wayne State University School of Medicine, Detroit, MI, USA.
Methods Mol Biol. 2018;1753:217-233. doi: 10.1007/978-1-4939-7720-8_15.
Retinal first-order neurons, photoreceptors, receive visual inputs and convert them to neural signals. The second-order neurons, bipolar cells then sort out the visual signals and encode them through multiple neural streams. Approximately 15 morphological types of bipolar cells have been identified, which are thought to encode different aspects of visual signals such as motion and color (Ichinose et al. J Neurosci 34(26):8761-8771, 2014; Euler et al. Nat Rev Neurosci 15(8):507-519, 2014). To investigate functional aspects of OFF bipolar cells, single cell recordings are preferred; however, bipolar cells in the mouse retina are small and hard to distinguish from other types of cells. Here, we describe our methodology and tips for immunohistochemistry and patch clamp recordings for analyzing light-evoked responses in each type of OFF bipolar cell.
视网膜的一级神经元,即光感受器,接收视觉输入并将其转换为神经信号。二级神经元,即双极细胞,随后对视觉信号进行分类,并通过多个神经流对其进行编码。已鉴定出大约15种形态类型的双极细胞,它们被认为对视觉信号的不同方面进行编码,如运动和颜色(市野濑等人,《神经科学杂志》34(26):8761 - 8771, 2014;欧拉等人,《自然评论神经科学》15(8):507 - 519, 2014)。为了研究OFF双极细胞的功能方面,单细胞记录是首选方法;然而,小鼠视网膜中的双极细胞很小,很难与其他类型的细胞区分开来。在这里,我们描述了用于免疫组织化学和膜片钳记录的方法和技巧,以分析每种类型的OFF双极细胞中的光诱发反应。