Abstract

The wound healing response of fibroblasts critically depends on the primary cilium, a sensory organelle protruding into the environment and comprising a stable axonemal structure. A characteristic marker for primary cilia is acetylation of axonemal tubulin. Although formation of primary cilia is under cell cycle control, the environmental cues affecting ciliation are not fully understood. Our purpose was, therefore, to study the impact of culture conditions on cilia formation in NIH3T3 fibroblasts. We quantified ciliation in different NIH3T3 sub-cell lines and culture conditions by immunodetection of primary cilia and counting. Quantitative Western blotting, qRT-PCR, and proliferation assays completed our investigation. We observed large differences between NIH3T3 sub-cell lines in their ability to generate acetylated primary cilia that correlated with cytoplasmic tubulin acetylation. We found no increased activity of the major tubulin deacetylase, HDAC6, but instead reduced expression of the α-tubulin acetyltransferase 1 (Atat1) as being causative. Our observations demonstrate that cells with reduced expression of Atat1 and tubulin acetylation proliferate faster, eventually displacing all other cells in the population. Expression of Atat1 and tubulin acetylation are therefore selective forces in cell competition.

Highlights

  • In the initial phase of wound healing, activated platelets trigger the release of growth factors, such as plateletderived growth factor (PDGF), that signal through the tyrosine kinase receptor PDGFRα located in the primary cilium membrane of fibroblasts causing re-entry into the cell cycle from quiescence and proliferation [3]

  • Our results demonstrate a significant impact of long-term cultivation conditions on intrinsic cellular features

  • alpha-tubulin acetyltransferase1 (Atat1) expression and tubulin acetylation that correlated with a decrease of acetylated primary cilia

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Summary

Introduction

Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. Fibroblasts are fundamental cells of the connective tissue. They maintain the structural integrity of the stroma by synthesizing the extracellular matrix. Fibroblasts play a critical role in wound healing and inflammatory diseases, like rheumatism, and fibrotic diseases [1]. Injury-stimulated migration of fibroblasts and their woundhealing capacity critically depend on a functional primary cilium [2]. In the initial phase of wound healing, activated platelets trigger the release of growth factors, such as plateletderived growth factor (PDGF), that signal through the tyrosine kinase receptor PDGFRα located in the primary cilium membrane of fibroblasts causing re-entry into the cell cycle from quiescence and proliferation [3]

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