Detail of DD25_Rod_CRX_R



Project
Title
Immunofluorescence image of photoreceptor-specific gene crx and Rhodopsin expressing in mouse iPS cells derived 3D retinal maintained containing 9cRA at differentiated day25
Description
NA
Release, Updated
2019-11-20
License
CC BY
Kind
Image data based on Experiment
File Formats
Data size
3.2 MB

Organism
M. musculus ( NCBI:txid10090 )
Strain(s)
-
Cell Line
-
Gene symbols
crx, Rhodopsin

Datatype
retina differentiation
Molecular Function (MF)
Biological Process (BP)
retinal rod cell differentiation ( GO:0060221 ) photoreceptor development ( GO:0008594 )
Cellular Component (CC)
photoreceptor cilium ( GO:0032391 )
Biological Imaging Method
XYZ Scale
XY:0.3125849 micrometer/pixel, Z: NA
T scale
-

Image Acquisition
Experiment type
Immunofluorescence
Microscope type
ConfocalMicroscope
Acquisition mode
FluorescenceCorrelationSpectroscopy
Contrast method
Fluorescence
Microscope model
ZEISS LSM700
Detector model
-
Objective model
-
Filter set
-

Summary of Methods
See details in Ueda et al. (2018) Biochem Biophys Res Commun, 495(4): 2595-2601.
Related paper(s)

Kaori Ueda, Akishi Onishi, Shin-Ichiro Ito, Makoto Nakamura, Masayo Takahashi (2018) Generation of three-dimensional retinal organoids expressing rhodopsin and S- and M-cone opsins from mouse stem cells., Biochemical and biophysical research communications, Volume 495, Number 4, pp. 2595-2601

Published in 2018 Jan 22 (Electronic publication in Dec. 20, 2017, midnight )

(Abstract) PURPOSE: Three-dimensional retinal organoids can be differentiated from embryonic stem cells/induced pluripotent stem cells (ES/iPS cells) under defined medium conditions. We modified the serum-free floating culture of embryoid body-like aggregates with quick reaggregation (SFEBq) culture procedure to obtain retinal organoids expressing more rod photoreceptors and S- and M-cone opsins. METHODS: Retinal organoids differentiated from mouse Nrl-eGFP iPS cells were cultured in various mediums during photoreceptor development. To promote rod photoreceptor development, organoids were maintained in media containing 9-cis retinoic acids (9cRA). To obtain retinal organoids with M-opsin expression, we cultured in medium with 1% fetal bovine serum (FBS) supplemented with T3, BMP4, and DAPT. Section immunohistochemistry was performed to visualize the expression of photoreceptor markers. RESULTS: In three-dimensional (3D) retinas exposed to 9cRA, rhodopsin was expressed earlier and S-cone opsins were suppressed. We could maintain 3D retinas up to DD 35 in culture media with 1% FBS. The 3D retinas expressed rhodopsin, S- and M-opsins, but most cone photoreceptors expressed either S- or M-opsins. CONCLUSION: By modifying culture conditions in the SFEBq protocol, we obtained rod-dominated 3D retinas and S- and M-opsin expressing 3D retinas.
(MeSH Terms)

Contact
Akishi Onishi , RIKEN , Center for Biosystems Dynamics Research , Laboratory for Retinal Regeneration
Contributors
Kaori Ueda, Akishi Onishi, Shin-ichiro Ito, Makoto Nakamura, Masayo Takahashi

OMERO Dataset
OMERO Project
Source