Abstract
Purpose :
Posterior capsular opacification (PCO) is a common complication of cataract surgery that appears to result from the proliferation and migration of residual lens epithelial cells (LECs) and their differentiation into either lens fiber cells (regeneration) or fibrotic myofibroblasts (epithelial to mesenchymal transition). However, immune cells also populate the capsular bag following cataract surgery, and in other tissues, fibrosis post-injury is driven by circulating fibrocytes. This study describes the initial characterization of a mouse model that will be used to determine the cellular origin of lens capsule-associated myofibroblasts generated in vivo following cataract surgery.
Methods :
Mice carrying a floxed H2b-mCherry allele (B6;129S-Gt(ROSA)26Sor<tm1.1Ksvo>/J) were obtained from the Jackson Laboratory and bred to MLR10Cre mice (STOCK Tg(Cryaa-cre)10Mlr/J) to create compound heterozygous MLR10Cre-H2b-mCherry mice. This genetic tool drives MLR10Cre expression, enabling permanent LEC labeling when bred with mice harboring StopFloxStop reporter alleles through the expression of the effector protein H2b-mCherry. For analysis, we collected eye tissues when mice were 2-5 months old and performed immunofluorescence staining to assess H2b-mCherry expression using a rabbit polyclonal antibody against H2b-mCherry, and sections were imaged on a confocal microscope.
Results :
MLR10Cre H2b-mCherry mice exhibit Cre-dependent expression of H2b-mCherry in both the nucleus and cytoplasm of lens epithelial and fiber cells in adult mice, while no H2B-mCherry signal was detected in control animals lacking MLR10Cre.
Conclusions :
MLR10Cre H2b-mCherry mice exhibit both nuclear and cytoplasmic expression of H2b-mCherry suggesting that these animals will be useful to track the fate of lens epithelial cells during their injury response in an animal model of cataract surgery. We are in the midst of validating the completeness of cellular labeling by whole lens/epithelial whole mount imaging and then will perform lens injuries to trace the fate of these cells during the lens injury response.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.