Lineage Tracing of Ascl1+ Retinal Progenitors

Ascl1^CreERT2 knock-in mice (Kim et al., 2011 (link)) were crossed with mTmG reporter mice (Jackson Laboratory, stock 007576) (Muzumdar et al., 2007 (link)). Pups were injected with 0.5 mg tamoxifen (Sigma) in 50 µl corn oil intraperitoneally at P4 to label the lineage of Ascl1+ retinal progenitors at the time of injection (rod photoreceptors, bipolar cells and Müller glia) with GFP. DM was injected intravitreally at P6 in one eye and the other eye was uninjected as a control. Eyes were collected at P21. GFP+ rod photoreceptors (in the ONL), bipolar cells (Otx2+ in the INL) and Müller glia (Otx2− in the INL) were blindly counted from random images of the central retina.

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Ueki Y., Wilken M.S., Cox K.E., Chipman L.B., Bermingham-McDonogh O, & Reh T.A. (2015). A transient wave of BMP signaling in the retina is necessary for Müller glial differentiation. Development (Cambridge, England), 142(3), 533-543.

Publication 2015

mTmG reporter mice tamoxifen

Cells Corn oil Eyes Glia Mice Retina Rod photoreceptors Tamoxifen

Corresponding Organization :

Other organizations : University of Washington

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Variable analysis

independent variables

Injection of 0.5 mg tamoxifen (Sigma) in 50 µl corn oil intraperitoneally at P4 to label the lineage of Ascl1+ retinal progenitors
Injection of DM intravitreally at P6 in one eye

dependent variables

Number of GFP+ rod photoreceptors (in the ONL)
Number of GFP+ bipolar cells (Otx2+ in the INL)
Number of GFP+ Müller glia (Otx2− in the INL)

control variables

Use of Ascl1^CreERT2 knock-in mice crossed with mTmG reporter mice
The other eye was uninjected as a control

controls

Positive control: Ascl1^CreERT2 knock-in mice crossed with mTmG reporter mice
Negative control: The other eye was uninjected

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