Gut epithelial Depdc5/mTORC1 signaling axis regulates mouse intestinal epithelial homeostasis
Objective To investigate the effect of over-activation of mTORC1 on mouse intestinal epithelial homeostasis by conditional knockout of Depdc5. Methods Mice were divided into Depdc5flox/flox group (control group), Depdc5flox/flox:Villin-Cre group (IKO group), rapamycin-Depdc5flox/flox group (rap-Ctrl...
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| Format: | Article |
| Language: | zho |
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Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College.
2022-08-01
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| Series: | Jichu yixue yu linchuang |
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| Online Access: | http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/1001-6325-2022-42-8-1213.pdf |
| _version_ | 1797365859756277760 |
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| author | ZHANG Xin-ge, MA Jie, WANG Qing-zhi, XIN Yue, YANG Chen-yan, XIONG Xi-wen |
| author_facet | ZHANG Xin-ge, MA Jie, WANG Qing-zhi, XIN Yue, YANG Chen-yan, XIONG Xi-wen |
| author_sort | ZHANG Xin-ge, MA Jie, WANG Qing-zhi, XIN Yue, YANG Chen-yan, XIONG Xi-wen |
| collection | DOAJ |
| description | Objective To investigate the effect of over-activation of mTORC1 on mouse intestinal epithelial homeostasis by conditional knockout of Depdc5. Methods Mice were divided into Depdc5flox/flox group (control group), Depdc5flox/flox:Villin-Cre group (IKO group), rapamycin-Depdc5flox/flox group (rap-Ctrl group) rapamycin-Depdc5flox/flox:Villin-Cre group (rap-IKO group). Western blot was used to detect the protein expression of DEPDC5 and the phosphorylation S6 (PS6) which is the downstream molecule of mTORC1 in the intestinal epithelium. qPCR was applied to detect the mRNA levels of Dclk1, Trpm5 and Muc2 in intestinal epithelial cells (IECs). Hematoxylin-eosin (HE) staining was used to observe the intestinal morphology changes. Immunohistochemical (IHC) staining and alcian blue staining were used to detect the number of tuft cells, Paneth cells, proliferative cells and goblet cells. Results The protein expression of PS6 in the small intestine and colon from the mice of IKO group was obviously up-regulated than that of the control group (0.957±0.028 vs. 0.598±0.041) (P＜0.05). The mRNA levels of Dclk1, Trpm5, and Muc2 were significantly decreased in the IKO group than those in the control group (P＜0.05). The numbers of tuft cells, Paneth cells and goblet cells in the IKO group were all significantly reduced than that in the control group. Nevertheless, the crypt depth in the IKO group was significantly higher than that of the control group (P＜0.05), and the number of proliferative cells was also increased in the IKO group(P＜0.05). Conclusions The over-activation of mTORC1 inhibits the differentiation of IECs and impairs the homeostasis of intestinal epithelial. |
| first_indexed | 2024-03-08T16:55:55Z |
| format | Article |
| id | doaj.art-72e306b4f04a4e2a85c48a900d0b3c2b |
| institution | Directory Open Access Journal |
| issn | 1001-6325 |
| language | zho |
| last_indexed | 2024-03-08T16:55:55Z |
| publishDate | 2022-08-01 |
| publisher | Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College. |
| record_format | Article |
| series | Jichu yixue yu linchuang |
| spelling | doaj.art-72e306b4f04a4e2a85c48a900d0b3c2b2024-01-05T02:36:08ZzhoInstitute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College.Jichu yixue yu linchuang1001-63252022-08-014281213121910.16352/j.issn.1001-6325.2022.08.1213Gut epithelial Depdc5/mTORC1 signaling axis regulates mouse intestinal epithelial homeostasisZHANG Xin-ge, MA Jie, WANG Qing-zhi, XIN Yue, YANG Chen-yan, XIONG Xi-wen01. School of Forensic Medicine; 2. Department of Human Anatomy & Histoembryology, School of Basic Medical Sciences, Xinxiang Medical University, Xinxiang 453000, ChinaObjective To investigate the effect of over-activation of mTORC1 on mouse intestinal epithelial homeostasis by conditional knockout of Depdc5. Methods Mice were divided into Depdc5flox/flox group (control group), Depdc5flox/flox:Villin-Cre group (IKO group), rapamycin-Depdc5flox/flox group (rap-Ctrl group) rapamycin-Depdc5flox/flox:Villin-Cre group (rap-IKO group). Western blot was used to detect the protein expression of DEPDC5 and the phosphorylation S6 (PS6) which is the downstream molecule of mTORC1 in the intestinal epithelium. qPCR was applied to detect the mRNA levels of Dclk1, Trpm5 and Muc2 in intestinal epithelial cells (IECs). Hematoxylin-eosin (HE) staining was used to observe the intestinal morphology changes. Immunohistochemical (IHC) staining and alcian blue staining were used to detect the number of tuft cells, Paneth cells, proliferative cells and goblet cells. Results The protein expression of PS6 in the small intestine and colon from the mice of IKO group was obviously up-regulated than that of the control group (0.957±0.028 vs. 0.598±0.041) (P＜0.05). The mRNA levels of Dclk1, Trpm5, and Muc2 were significantly decreased in the IKO group than those in the control group (P＜0.05). The numbers of tuft cells, Paneth cells and goblet cells in the IKO group were all significantly reduced than that in the control group. Nevertheless, the crypt depth in the IKO group was significantly higher than that of the control group (P＜0.05), and the number of proliferative cells was also increased in the IKO group(P＜0.05). Conclusions The over-activation of mTORC1 inhibits the differentiation of IECs and impairs the homeostasis of intestinal epithelial.http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/1001-6325-2022-42-8-1213.pdfdepdc5|mtorc1|iecs|rapamycin |
| spellingShingle | ZHANG Xin-ge, MA Jie, WANG Qing-zhi, XIN Yue, YANG Chen-yan, XIONG Xi-wen Gut epithelial Depdc5/mTORC1 signaling axis regulates mouse intestinal epithelial homeostasis Jichu yixue yu linchuang depdc5|mtorc1|iecs|rapamycin |
| title | Gut epithelial Depdc5/mTORC1 signaling axis regulates mouse intestinal epithelial homeostasis |
| title_full | Gut epithelial Depdc5/mTORC1 signaling axis regulates mouse intestinal epithelial homeostasis |
| title_fullStr | Gut epithelial Depdc5/mTORC1 signaling axis regulates mouse intestinal epithelial homeostasis |
| title_full_unstemmed | Gut epithelial Depdc5/mTORC1 signaling axis regulates mouse intestinal epithelial homeostasis |
| title_short | Gut epithelial Depdc5/mTORC1 signaling axis regulates mouse intestinal epithelial homeostasis |
| title_sort | gut epithelial depdc5 mtorc1 signaling axis regulates mouse intestinal epithelial homeostasis |
| topic | depdc5|mtorc1|iecs|rapamycin |
| url | http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/1001-6325-2022-42-8-1213.pdf |
| work_keys_str_mv | AT zhangxingemajiewangqingzhixinyueyangchenyanxiongxiwen gutepithelialdepdc5mtorc1signalingaxisregulatesmouseintestinalepithelialhomeostasis |