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Transpupillary Two-Photon In Vivo Imaging of the Mouse Retina
Published on: February 13, 2021
Expression of Cre recombinase in retinal Müller cells
Yumi Ueki1, John D Ash, Meili Zhu
1Oklahoma Center for Neuroscience, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.
Purpose:
In an effort to generate inducible RPE-specific Cre mice using a 3.0-kb human vitelliform macular dystrophy-2 (VMD2) promoter, we identified a mouse line with unanticipated Cre activity in the neural retina, including Müller glial cells. Müller cells play important roles in the function and maintenance of the retina, and this mouse line would be potentially useful for conditional gene targeting in Müller glia. We therefore characterized the timing, inducibility, and cell specificity of Cre expression, as well as Müller cell-specific efficiency of Cre-mediated recombination in this mouse line.
Methods:
Transgenic mice carrying cassettes of human P(VMD2)-rtTA and TRE-cre were generated. Cre expression was characterized using a Cre-activatable lacZ reporter mouse line (R26R) and a floxed interleukin six signal transducing receptor (gp130) mouse line.
Results:
beta-Galactosidase (beta-gal) assay and immunohistochemical analysis of VMD2-cre/R26R double transgenic mice indicated that Cre activity was detected in cells located in the inner nuclear layer, with prominent expression of beta-gal in Müller cells. Cre activity was also detected in photoreceptors in the outer nuclear layer. PCR analysis demonstrated that Cre-mediated recombination initiated by embryonic day 15. Immunohistochemical analysis indicated that Cre-mediated deletion of floxed gp130 gene occurred in 52% of the retinal Müller cells. Retinal function and morphology were normal in 10-month-old VMD2-cre mice.
Conclusion:
We generated a transgenic cre mouse that is useful to study gene activation and inactivation in retinal Müller cells.
Insights
A novel transgenic mouse model exhibits Cre recombinase activity in retinal Müller glial cells, enabling conditional gene targeting for studying retinal diseases. This tool is valuable for investigating Müller glia function and maintenance.
Area of Science:
- Retinal biology
- Gene targeting
- Transgenic mouse models
Background:
- Müller glial cells are crucial for retinal function and maintenance.
- Conditional gene targeting is essential for studying specific cell functions in vivo.
- The vitelliform macular dystrophy-2 (VMD2) promoter is a target for RPE-specific gene expression.
Purpose of the Study:
- To develop an inducible RPE-specific Cre mouse line using the VMD2 promoter.
- To characterize a mouse line with unexpected Cre activity in Müller glial cells.
- To evaluate the utility of this mouse line for conditional gene targeting in Müller glia.
Main Methods:
- Generation of transgenic mice with human P(VMD2)-rtTA and TRE-cre cassettes.
- Cre expression analysis using a Cre-activatable lacZ reporter (R26R) mouse line.
- Assessment of Cre-mediated recombination via a floxed interleukin-6 signal transducer (gp130) mouse line.
Main Results:
- Cre activity was observed in Müller cells and photoreceptors of the neural retina.
- Cre-mediated recombination initiated by embryonic day 15.
- Successful deletion of the floxed gp130 gene in 52% of retinal Müller cells.
- Normal retinal function and morphology in VMD2-cre mice up to 10 months of age.
Conclusions:
- A transgenic Cre mouse model with activity in retinal Müller cells was successfully generated.
- This VMD2-driven Cre mouse line is a valuable tool for studying gene activation and inactivation in Müller glia.
- The model facilitates research into Müller cell biology and its role in retinal health and disease.
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