Altered Cell Clusters and Upregulated Aqp1 in Connexin 50 Knockout Lens Epithelium

Area of Science:

• Ophthalmology
• Cell Biology
• Genomics

Background:

• Lens epithelial cells (LECs) are crucial for maintaining lens homeostasis and growth.
• Connexin 50 (Cx50) plays a vital role in lens development and function.
• Cx50 deficiency is associated with cataracts and reduced lens size.

Purpose of the Study:

• To characterize heterogeneity and cell clusters in postnatal LECs.
• To identify downstream targets of Cx50 in regulating lens homeostasis and growth.
• To determine differentially expressed genes (DEGs) in Cx50 knockout (Cx50KO) LECs to understand cataract mechanisms.

Main Methods:

• Single-cell RNA sequencing (scRNA-seq) was performed on LECs from Cx50KO and wild-type (WT) mice.
• Differentially expressed genes (DEGs) were identified.
• Quantitative real-time PCR (RT-qPCR) and Western blot analysis were used to validate selected DEGs, including aquaporin 1 (Aqp1).

Main Results:

• scRNA-seq revealed thousands of gene expression profiles in LECs.
• Cx50KO LECs exhibited numerous DEGs compared to WT controls, including growth regulators and water channel genes.
• Significantly upregulated Aqp1 gene and protein expression was confirmed in Cx50KO LECs.

Conclusions:

• LECs display intrinsic heterogeneity across different cell clusters, influencing lens homeostasis and growth.
• Upregulated Aqp1 in Cx50KO LECs suggests a critical role for both Cx50 and AQP1 in regulating water homeostasis.
• These findings provide novel insights into the molecular mechanisms underlying Cx50-related cataract and lens growth defects.