A Qualitative Change in the Transcriptome Occurs after the First Cell Cycle and Coincides with Lumen Establishment during MDCKII Cystogenesis
Tianfang Wang,
Sang-Ho Kwon,
Xiao Peng,
Severine Urdy,
Zefu Lu,
Robert J. Schmitz,
Stephen Dalton,
Keith E. Mostov,
Shaying Zhao
Affiliations
Tianfang Wang
Department of Biochemistry and Molecular Biology, Institute of Bioinformatics, University of Georgia, Athens, GA 30602, USA
Sang-Ho Kwon
Department of Anatomy, University of California, San Francisco, CA 94143-2140, USA; Department of Cellular Biology and Anatomy, Augusta University, Medical College of Georgia, 1460 Laney Walker Boulevard, CB2820A, Augusta, GA 30912, USA
Xiao Peng
Department of Anatomy, University of California, San Francisco, CA 94143-2140, USA
Severine Urdy
Department of Anatomy, University of California, San Francisco, CA 94143-2140, USA
Zefu Lu
Department of Genetics, University of Georgia, Athens, GA 30602, USA
Robert J. Schmitz
Department of Genetics, University of Georgia, Athens, GA 30602, USA
Stephen Dalton
Department of Biochemistry and Molecular Biology, Institute of Bioinformatics, University of Georgia, Athens, GA 30602, USA
Keith E. Mostov
Department of Anatomy, University of California, San Francisco, CA 94143-2140, USA
Shaying Zhao
Department of Biochemistry and Molecular Biology, Institute of Bioinformatics, University of Georgia, Athens, GA 30602, USA; Corresponding author
Summary: Madin-Darby canine kidney II (MDCKII) cells are widely used to study epithelial morphogenesis. To better understand this process, we performed time course RNA-seq analysis of MDCKII 3D cystogenesis, along with polarized 2D cells for comparison. Our study reveals a biphasic change in the transcriptome that occurs after the first cell cycle and coincides with lumen establishment. This change appears to be linked to translocation of β-catenin, supported by analyses with AVL9- and DENND5A-knockdown clones, and regulation by HNF1B, supported by ATAC-seq study. These findings indicate a qualitative change model for transcriptome remodeling during epithelial morphogenesis, leading to cell proliferation decrease and cell polarity establishment. Furthermore, our study reveals that active mitochondria are retained and chromatin accessibility decreases in 3D cysts but not in 2D polarized cells. This indicates that 3D culture is a better model than 2D culture for studying epithelial morphogenesis.
