Fibroblast Growth Factor Receptor 2c Signaling Is Required for Intestinal Cell Differentiation in Zebrafish

PLOS ONE, Dec 2019

Background There are four cell lineages derived from intestinal stem cells that are located at the crypt and villus in the mammalian intestine the non-secretory absorptive enterocytes, and the secretory cells, which include mucous-secreting goblet cells, regulatory peptide-secreting enteroendocrine cells and antimicrobial peptide-secreting Paneth cells. Although fibroblast growth factor (Fgf) signaling is important for cell proliferation and differentiation in various tissues, its role in intestinal differentiation is less well understood. Methodology/Principal Findings We used a loss of function approach to investigate the importance of Fgf signaling in intestinal cell differentiation in zebrafish; abnormal differentiation of goblet cells was observed when Fgf signaling was inhibited using SU5402 or in the Tg(hsp70ldnfgfr1-EGFP) transgenic line. We identified Fgfr2c as an important receptor for cell differentiation. The number of goblet cells and enteroendocrine cells was reduced in fgfr2c morphants. In addition to secretory cells, enterocyte differentiation was also disrupted in fgfr2c morphants. Furthermore, proliferating cells were increased in the morphants. Interestingly, the loss of fgfr2c expression repressed secretory cell differentiation and increased cell proliferation in the mibta52b mutant that had defective Notch signaling. Conclusions/Significance In conclusion, we found that Fgfr2c signaling derived from mesenchymal cells is important for regulating the differentiation of zebrafish intestine epithelial cells by promoting cell cycle exit. The results of Fgfr2c knockdown in mibta52b mutants indicated that Fgfr2c signaling is required for intestinal cell differentiation. These findings provide new evidences that Fgf signaling is required for the differentiation of intestinal cells in the zebrafish developing gut.

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Fibroblast Growth Factor Receptor 2c Signaling Is Required for Intestinal Cell Differentiation in Zebrafish

Wang W-P (2013) Fibroblast Growth Factor Receptor 2c Signaling Is Required for Intestinal Cell Differentiation in Zebrafish. PLoS ONE 8(3): e58310. doi:10.1371/journal.pone.0058310 Fibroblast Growth Factor Receptor 2c Signaling Is Required for Intestinal Cell Differentiation in Zebrafish Da-Wei Liu 0 Su-Mei Tsai 0 Bih-Fen Lin 0 Yun-Jin Jiang 0 Wen-Pin Wang 0 Shree Ram Singh, National Cancer Institute, United States of America 0 1 Institute of Medical Sciences, Tzu-Chi University , Hualien, Taiwan , 2 Department of Laboratory Medicine and Biotechnology, Tzu-Chi University , Hualien, Taiwan , 3 Division of Molecular and Genomic Medicine, National Health Research Institutes , Zhunan Town, Miaoli County , Taiwan , 4 Department of Molecular Biology and Human Genetics, Tzu-Chi University , Hualien , Taiwan Background: There are four cell lineages derived from intestinal stem cells that are located at the crypt and villus in the mammalian intestine the non-secretory absorptive enterocytes, and the secretory cells, which include mucous-secreting goblet cells, regulatory peptide-secreting enteroendocrine cells and antimicrobial peptide-secreting Paneth cells. Although fibroblast growth factor (Fgf) signaling is important for cell proliferation and differentiation in various tissues, its role in intestinal differentiation is less well understood. Methodology/Principal Findings: We used a loss of function approach to investigate the importance of Fgf signaling in intestinal cell differentiation in zebrafish; abnormal differentiation of goblet cells was observed when Fgf signaling was inhibited using SU5402 or in the Tg(hsp70ldnfgfr1-EGFP) transgenic line. We identified Fgfr2c as an important receptor for cell differentiation. The number of goblet cells and enteroendocrine cells was reduced in fgfr2c morphants. In addition to secretory cells, enterocyte differentiation was also disrupted in fgfr2c morphants. Furthermore, proliferating cells were increased in the morphants. Interestingly, the loss of fgfr2c expression repressed secretory cell differentiation and increased cell proliferation in the mibta52b mutant that had defective Notch signaling. Conclusions/Significance: In conclusion, we found that Fgfr2c signaling derived from mesenchymal cells is important for regulating the differentiation of zebrafish intestine epithelial cells by promoting cell cycle exit. The results of Fgfr2c knockdown in mibta52b mutants indicated that Fgfr2c signaling is required for intestinal cell differentiation. These findings provide new evidences that Fgf signaling is required for the differentiation of intestinal cells in the zebrafish developing gut. - Funding: This work was supported by grants from the National Science Council (NSC100-2311-B-320-001), and Tzu Chi University (TCMRC-P-99013-01). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. In adult mammals, the epithelium of the small intestine comprises two structures: finger-like villi and pocket-like crypts of Lieberk uhn. Intestinal stem cells are located at the bottom of the crypt. Crypts also contain transit amplifying progenitor cells. These proliferating cells differentiate, then migrate to villi and are removed at the top of the villi by apoptosis. There are four cell lineages that derive from intestinal stem cells: the non-secretory absorptive enterocytes, and secretory cells, which include mucoussecreting goblet cells, regulatory peptide-secreting enteroendocrine cells, and antimicrobial peptide-secreting Paneth cells [1,2,3,4]. It has been reported that, unlike mammals, zebrafish do not possess crypts of Lieberku hn or Paneth cells [5]. Many signaling molecules regulate stem cell self-renewal, proliferation, and differentiation in the intestines [6,7]. The Wnt pathway is important in controlling crypt cell proliferation. The crypt precursors of Tcf4 null mice exhibit decreased cell proliferation, and comprise various differentiated cells [8]. However, in mice that lack Apc expression (APCmin), crypt cells exhibit greater proliferation than they do in Tcf4 null mice, and in the deficient mice, these cells only differentiate to form Paneth cells [9,10]. In Apc mutant zebrafish (Apcmcr), the enterocyte differentiation marker, intestinal fatty acid binding protein (ifabp), was failed to express [11,12]. In bone morphogenetic protein receptor 1a (Bmpr1a) mutant mice or Noggin transgenic mice, the expansion of proliferating cells in the crypt results in intestinal polyposis [13,14]. Three secretory cells are also reduced in Bmpr1a mutant mice [15]. Interestingly, Wnt signaling is highly activated in these Bmp pathway deficient mice. Additionally, Notch signaling is important for cell lineage commitment and proliferation. Notch1 and Notch2 double knockout mice exhibit complete conversion of proliferating cryp (...truncated)


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Da-Wei Liu, Su-Mei Tsai, Bih-Fen Lin, Yun-Jin Jiang, Wen-Pin Wang. Fibroblast Growth Factor Receptor 2c Signaling Is Required for Intestinal Cell Differentiation in Zebrafish, PLOS ONE, 2013, Volume 8, Issue 3, DOI: 10.1371/journal.pone.0058310