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

CIL:13523*  Cite 
Description

A confocal Z-stack of neurons in postnatal hippocampus, expressing a GFP-tagged knockdown construct for the microRNA miR-137 (green, sh-miR-137) and immunostained to localize doublecortin (red), a marker of neuronal differentiation, and labeled with Dapi (blue), to determine the function of this microRNA in vivo. miR-137 regulates a core transcription factor in stem cell renewal, and data from these experiments suggest that high levels of miR-137promotes neuronal stem cell proliferation, but repressed neuronal differentiation. This image is original data from Fig. 5 of Szulwach et al.(2010) Cross talk between microRNA and epigenetic regulation in adult neurogenesis, J. Cell Biol. Vol. 189:127–141.

Technical Details

A retroviral vector containing sh-miR-137 under a U6 promoter and EGFP under a chicken actin promoter was prepared, and the virus containing the construct was injected into the dentate gyrus of adult mice in vivo. After 1 week, animals were anesthetized, perfused with 4% paraformaldehyde, brains removed and postfixed, equilibrated in 30% sucrose, and sectioned at a thickness of 40µm with a sliding microtome. Infected cells were localized using chicken-α-GFP (Invitrogen) and rabbit-α-DCX (Cell Signaling Technology), secondaries were α-chicken Alexa Fluor 488 and goat-α-rabbit Alexa Fluor 568 (both from Invitrogen). Confocal z-stacks were acquired at 1 µm steps using a Nikon TE2000 with equipped with a spinning disc confocal with an oil immersion 40X 1.3 NA objective (Zeiss). For additional details see: J. Cell Biol. Vol. 189:127–141.

Biological Sources
NCBI Organism Classification
Mus musculus
Cell Type
neuronal stem cell
Cellular Component
nucleus
Attribution
Names
Keith E. Szulwach
Xuekun Li
Richard D. Smrt
Yujing Li
Yuping Luo
Li Lin
Nicholas J. Santistevan
Wendi Li
Xinyu Zhao
Peng Jin
Pubmed
PMID:20368621
Link
http://jcb-dataviewer.rupress.org/jcb/browse/...
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL13523
Archival Resource Key (ARK)
ark:/b7295/w9cil13523
Grouping This image is part of a group.
Sample Preparation
Methods
formaldehyde fixed tissue
permeabilized tissue
Relation To Intact Cell
microtome-sectioned tissue
Dimensions
Spatial Axis Image Size Pixel Size
X 1024px ——
Y 1024px ——
Z 8px 1µm