Margaret I. Davis (2011) CIL:14971, Mus musculus, interneuron, hippocampal granule cell, pyramidal cell. CIL. Dataset

Distribution of interneurons expressing EGFP from the 5HT3 receptor promoter (Tg(Htr3a-EGFP)DH30Gsat, www.gensat.org) in the dorsal hippocampus colabelled for the CB1 cannabinoid receptor (red) and counterstained with DAPI (blue) to show the cell layers. EGFP expression was amplified with chicken anti-GFP (Abcam, 1:2000); cell bodies and fibers are present throughout all layers of the hippocampus but enriched in the hilus and stratum lacunosum moleculare (see associated images). CB1 immunoreactivity (L15 rabbit polyclonal 1:200, K. Mackie) is prominent in the terminals of basket cells synapsing in the pyramidal cell layer. CB1 is also enriched in axons with distinct intensities in the inner and outer molecular layer of the dentate gyrus. CB immunoreactivity is also present in the stratum radiatum and stratum lacunosum moleculare.

Section (40 um) was mounted with Fluoromount G and imaged with a 5X, 0.15 NA Achroplan objective on a Zeiss Axiovert microscope. AF568 was excited at 562/40 nm and emission collected at 624/40 nm with Semrock filters and a single band dichroic (530-585 nm reflected, 601-800 nm transmitted). AF488 was excited at 482/35 nm and emission collected at 536/40 nm with Semrock filters and a single band dichroic (446-499 nm reflected, 513-735 nm transmitted). DAPI was excited at 350/50 nm and emission collected at 460/50 nm with a CLP400 dichroic (Chroma). Single images were captured for each channel using a Zeiss Axiocam MR CCD and pseudo-colored using Axiovision. Linear background subtraction was used on the presented image. See Davis and Puhl 2011 (Pubmed ID 21305052) for a detailed staining protocol. This image is part of a series characterizing EGFP expression from the 5HT3 promoter throughout the brain.

5HT3受体启动子表达的EGFP标记的内细胞群在大脑皮层中的分布（Tg(Htr3a-EGFP)DH30Gsat，www.gensat.org），与CB1大麻素受体（红色）共标记，并经DAPI染色（蓝色）以显示细胞层。EGFP表达通过鸡抗GFP抗体（Abcam，1:2000）进行增强；细胞体和纤维遍布海马的所有层次，但在海马门和分子层（参见相关图像）中富集。CB1免疫反应性（L15兔多克隆抗体1:200，K. Mackie）在篮状细胞与锥体层突触的突触末端明显。CB1在大脑回的内外分子层中也富集，具有不同的强度。CB免疫反应性也存在于辐射层和分子层。切片（40微米）使用Fluoromount G封片，并在蔡司Axiovert显微镜上使用5倍、0.15数值孔径的Achroplan物镜进行成像。AF568在562/40纳米处激发，并通过Semrock滤光片和单色二向色镜（530-585纳米反射，601-800纳米透过）收集发射光，发射光收集于624/40纳米。AF488在482/35纳米处激发，并通过Semrock滤光片和单色二向色镜（446-499纳米反射，513-735纳米透过）收集发射光，发射光收集于536/40纳米。DAPI在350/50纳米处激发，并通过CLP400二向色镜（Chroma）收集发射光，发射光收集于460/50纳米。使用蔡司Axiocam MR CCD捕获每个通道的单个图像，并使用Axiovision进行伪彩色处理。在展示的图像上使用线性背景减法。详见Davis和Puhl 2011年发表的文章（PubMed ID 21305052）以获取详细的染色方案。此图像是描述5HT3启动子在整个大脑中EGFP表达的一组图像之一。