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

doi:10.1016/j.isci.2020.101629

A Qualitative Change in the Transcriptome Occurs after the First Cell Cycle and Coincides with Lumen Establishment during MDCKII Cystogenesis

iScience. 2020 Sep 29;23(10):101629. doi: 10.1016/j.isci.2020.101629. eCollection 2020 Oct 23.

Authors

Tianfang Wang¹, Sang-Ho Kwon^{2

3}, Xiao Peng², Severine Urdy², Zefu Lu⁴, Robert J Schmitz⁴, Stephen Dalton¹, Keith E Mostov², Shaying Zhao¹

Affiliations

¹ Department of Biochemistry and Molecular Biology, Institute of Bioinformatics, University of Georgia, Athens, GA 30602, USA.
² Department of Anatomy, University of California, San Francisco, CA 94143-2140, USA.
³ Department of Cellular Biology and Anatomy, Augusta University, Medical College of Georgia, 1460 Laney Walker Boulevard, CB2820A, Augusta, GA 30912, USA.
⁴ Department of Genetics, University of Georgia, Athens, GA 30602, USA.

Abstract

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.

Keywords: Developmental Biology; Embryology; Transcriptomics.

Abstract

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