Xinyuan Wang, Xinyi Li, Yunsheng Wang, Zhongmei Ren, Xueqing Du, Jing Gao, Guangdong Ji, Zhenhui Liu
{"title":"Nkx1.2 基因缺失会降低斑马鱼的脂肪产量","authors":"Xinyuan Wang, Xinyi Li, Yunsheng Wang, Zhongmei Ren, Xueqing Du, Jing Gao, Guangdong Ji, Zhenhui Liu","doi":"10.1002/oby.24043","DOIUrl":null,"url":null,"abstract":"<div>\n \n \n <section>\n \n <h3> Objective</h3>\n \n <p>This study aimed to investigate the role of Nkx1-2, a transcription factor with the NK homeobox domain, in the regulation of fat production.</p>\n </section>\n \n <section>\n \n <h3> Methods</h3>\n \n <p>Gene expression was analyzed using quantitative real-time polymerase chain reaction or transcriptome sequencing. CRISPR/Cas9 technology was employed to generate <i>nkx1.2</i> knockout zebrafish and <i>nkx1.2</i>-deleted 3T3-L1 cells. Lipid droplet production in zebrafish larvae was visually quantified using Nile red staining, whereas lipid droplets in 3T3-L1 cells were stained with Oil red O. The binding of Nkx1-2 to the promoter was verified through an electrophoretic mobility shift assay experiment.</p>\n </section>\n \n <section>\n \n <h3> Results</h3>\n \n <p>Nkx1-2 plays crucial roles in the regulation of fat production in zebrafish. Knockout of <i>nkx1.2</i> in zebrafish leads to weight loss, accompanied by significantly reduced lipid droplet production and decreased visceral and liver fat content. Furthermore, genes related to lipid biosynthesis are significantly downregulated. In 3T3-L1 preadipocytes, Nkx1-2 induces differentiation into mature adipocytes by binding to the <i>cebpa</i> promoter, thereby activating its transcription. Additionally, the expression of <i>nkx1.2</i> is regulated by the p38 MAPK, JNK, or Smad2/3 signaling pathways in 3T3-L1 cells.</p>\n </section>\n \n <section>\n \n <h3> Conclusions</h3>\n \n <p>Our findings suggest that Nkx1-2 functions as a positive regulator of fat production, playing a critical role in adipocyte differentiation and lipid biosynthesis.</p>\n </section>\n </div>","PeriodicalId":215,"journal":{"name":"Obesity","volume":"32 7","pages":"1315-1328"},"PeriodicalIF":4.2000,"publicationDate":"2024-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Nkx1.2 deletion decreases fat production in zebrafish\",\"authors\":\"Xinyuan Wang, Xinyi Li, Yunsheng Wang, Zhongmei Ren, Xueqing Du, Jing Gao, Guangdong Ji, Zhenhui Liu\",\"doi\":\"10.1002/oby.24043\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div>\\n \\n \\n <section>\\n \\n <h3> Objective</h3>\\n \\n <p>This study aimed to investigate the role of Nkx1-2, a transcription factor with the NK homeobox domain, in the regulation of fat production.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Methods</h3>\\n \\n <p>Gene expression was analyzed using quantitative real-time polymerase chain reaction or transcriptome sequencing. CRISPR/Cas9 technology was employed to generate <i>nkx1.2</i> knockout zebrafish and <i>nkx1.2</i>-deleted 3T3-L1 cells. Lipid droplet production in zebrafish larvae was visually quantified using Nile red staining, whereas lipid droplets in 3T3-L1 cells were stained with Oil red O. The binding of Nkx1-2 to the promoter was verified through an electrophoretic mobility shift assay experiment.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Results</h3>\\n \\n <p>Nkx1-2 plays crucial roles in the regulation of fat production in zebrafish. Knockout of <i>nkx1.2</i> in zebrafish leads to weight loss, accompanied by significantly reduced lipid droplet production and decreased visceral and liver fat content. Furthermore, genes related to lipid biosynthesis are significantly downregulated. In 3T3-L1 preadipocytes, Nkx1-2 induces differentiation into mature adipocytes by binding to the <i>cebpa</i> promoter, thereby activating its transcription. 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Nkx1.2 deletion decreases fat production in zebrafish
Objective
This study aimed to investigate the role of Nkx1-2, a transcription factor with the NK homeobox domain, in the regulation of fat production.
Methods
Gene expression was analyzed using quantitative real-time polymerase chain reaction or transcriptome sequencing. CRISPR/Cas9 technology was employed to generate nkx1.2 knockout zebrafish and nkx1.2-deleted 3T3-L1 cells. Lipid droplet production in zebrafish larvae was visually quantified using Nile red staining, whereas lipid droplets in 3T3-L1 cells were stained with Oil red O. The binding of Nkx1-2 to the promoter was verified through an electrophoretic mobility shift assay experiment.
Results
Nkx1-2 plays crucial roles in the regulation of fat production in zebrafish. Knockout of nkx1.2 in zebrafish leads to weight loss, accompanied by significantly reduced lipid droplet production and decreased visceral and liver fat content. Furthermore, genes related to lipid biosynthesis are significantly downregulated. In 3T3-L1 preadipocytes, Nkx1-2 induces differentiation into mature adipocytes by binding to the cebpa promoter, thereby activating its transcription. Additionally, the expression of nkx1.2 is regulated by the p38 MAPK, JNK, or Smad2/3 signaling pathways in 3T3-L1 cells.
Conclusions
Our findings suggest that Nkx1-2 functions as a positive regulator of fat production, playing a critical role in adipocyte differentiation and lipid biosynthesis.
期刊介绍：
Obesity is the official journal of The Obesity Society and is the premier source of information for increasing knowledge, fostering translational research from basic to population science, and promoting better treatment for people with obesity. Obesity publishes important peer-reviewed research and cutting-edge reviews, commentaries, and public health and medical developments.
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