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*CIL – Cell Image Library accession number. Please use this to reference an image.

CIL:16244*  Cite 
Description

Distribution of hippocampal neurons expressing EGFP from the Nr4a1/Nur77 promoter (Tg(Nr4a1-EGFP)GY139Gsat, www.gensat.org) in hippocampal CA1 colabelled for the vesicular glutamate transporter (VGluT1, red, Invitrogen polyclonal, 1:1000) and stained with DAPI (blue) to show cell layers. EGFP expression was amplified with chicken anti-GFP (Abcam, 1:2000). VGluT1 labels excitatory terminals synapsing in the molecular layer of the dentate gyrus, mossy fiber terminals in CA3 and dendritic fields of the pyramidal cells in stratum orien and stratum radiatum. Nr4a1-EGFP is heavily expressed by CA1 pyramidal cells and to a lesser extent in CA3 pyramidal cells with sparse cells in the dentate gyrus (see associated images). Section (40 um, sagittal) was mounted with Fluoromount G and imaged with a 5X, 0.15 NA 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 and images of EGFP expression in the striatum and amygdala in this strain. This image is part of a series characterizing EGFP expression from the Nr4a1 promoter throughout the brain.

Biological Sources
NCBI Organism Classification
Mus musculus
Cell Type
pyramidal cell
hippocampal granule cell
Cellular Component
axon
dendrite
Attribution
Names
Margaret I. Davis
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL16244
Archival Resource Key (ARK)
ark:/b7295/w9cil16244
Grouping This image is part of a group.
Imaging
Image Type
recorded image
Image Mode
fluorescence microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
compartmentalization of stain or label
distribution of a specific protein
Visualization Methods
4',6-diamidino-2-phenylindole (DAPI)
EGFP
Alexa Fluor 488
Alexa Fluor 568
Data Qualifiers
raw, unprocessed data
Sample Preparation
Methods
formaldehyde fixed tissue
Relation To Intact Cell
vibratome-sectioned tissue
Dimensions
Spatial Axis Image Size Pixel Size
X 1300px 2.03µm
Y 1030px 2.03µm