Role of mitochondrial fission-related genes in mitochondrial morphology and energy metabolism in mouse embryonic stem cells
Mitochondria, the major organelles that produce energy for cell survival and function, dynamically change their morphology via fusion and fission, a process called mitochondrial dynamics. The details of the underlying mechanism of mitochondrial dynamics have not yet been elucidated. Here, we aimed t...
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| Format: | Article |
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Elsevier
2020-09-01
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| Series: | Redox Biology |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2213231720306340 |
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| author | Bong Jong Seo Joonhyuk Choi Hyeonwoo La Omer Habib Youngsok Choi Kwonho Hong Jeong Tae Do |
| author_facet | Bong Jong Seo Joonhyuk Choi Hyeonwoo La Omer Habib Youngsok Choi Kwonho Hong Jeong Tae Do |
| author_sort | Bong Jong Seo |
| collection | DOAJ |
| description | Mitochondria, the major organelles that produce energy for cell survival and function, dynamically change their morphology via fusion and fission, a process called mitochondrial dynamics. The details of the underlying mechanism of mitochondrial dynamics have not yet been elucidated. Here, we aimed to investigate the function of mitochondrial fission genes in embryonic stem cells (ESCs). To this end, we generated homozygous knockout ESC lines, namely, Fis1−/−, Mff−/−, and Dnm1l−/− ESCs, using the CRISPR-Cas9 system. Interestingly, the Fis1−/−, Mff−/−, and Dnm1l−/− ESCs showed normal morphology, self-renewal, and the ability to differentiate into all three germ layers in vitro. However, transmission electron microscopy showed a significant increase in the cytoplasm to nucleus ratio and mitochondrial elongation in Dnm1l−/− ESCs, which was due to incomplete fission. To assess the change in metabolic energy, we analyzed oxidative phosphorylation (OXPHOS), glycolysis, and the intracellular ATP concentration. The ESC knockout lines showed an increase in OXPHOS, decrease in glycolysis, and an increase in intracellular ATP concentration, which was related to mitochondrial elongation. In particular, the Dnm1l knockout most significantly affected mitochondrial morphology, energy metabolism, and ATP production in ESCs. Furthermore, RNA sequencing and gene ontology analysis showed that the differentially expressed genes in Mff-/- ESCs were distinct from those in Dnm1l−/- or Fis1−/− ESCs. In total, five metabolism-related genes, namely, Aass, Cdo1, Cyp2b23, Nt5e, and Pck2, were expressed in all three knockout ESC lines, and three of them were associated with regulation of ATP generation. |
| first_indexed | 2024-12-21T01:24:20Z |
| format | Article |
| id | doaj.art-b928fe1ab4e6487e9c92f9101c24311a |
| institution | Directory Open Access Journal |
| issn | 2213-2317 |
| language | English |
| last_indexed | 2024-12-21T01:24:20Z |
| publishDate | 2020-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Redox Biology |
| spelling | doaj.art-b928fe1ab4e6487e9c92f9101c24311a2022-12-21T19:20:34ZengElsevierRedox Biology2213-23172020-09-0136101599Role of mitochondrial fission-related genes in mitochondrial morphology and energy metabolism in mouse embryonic stem cellsBong Jong Seo0Joonhyuk Choi1Hyeonwoo La2Omer Habib3Youngsok Choi4Kwonho Hong5Jeong Tae Do6Department of Stem Cell and Regenerative Biotechnology, Konkuk Institute of Technology, Konkuk University, Seoul, Republic of KoreaDepartment of Stem Cell and Regenerative Biotechnology, Konkuk Institute of Technology, Konkuk University, Seoul, Republic of KoreaDepartment of Stem Cell and Regenerative Biotechnology, Konkuk Institute of Technology, Konkuk University, Seoul, Republic of KoreaDepartment of Chemistry, Hanyang University, Seoul, Republic of KoreaDepartment of Stem Cell and Regenerative Biotechnology, Konkuk Institute of Technology, Konkuk University, Seoul, Republic of KoreaDepartment of Stem Cell and Regenerative Biotechnology, Konkuk Institute of Technology, Konkuk University, Seoul, Republic of KoreaDepartment of Stem Cell and Regenerative Biotechnology, Konkuk Institute of Technology, Konkuk University, Seoul, Republic of Korea; Corresponding author.Mitochondria, the major organelles that produce energy for cell survival and function, dynamically change their morphology via fusion and fission, a process called mitochondrial dynamics. The details of the underlying mechanism of mitochondrial dynamics have not yet been elucidated. Here, we aimed to investigate the function of mitochondrial fission genes in embryonic stem cells (ESCs). To this end, we generated homozygous knockout ESC lines, namely, Fis1−/−, Mff−/−, and Dnm1l−/− ESCs, using the CRISPR-Cas9 system. Interestingly, the Fis1−/−, Mff−/−, and Dnm1l−/− ESCs showed normal morphology, self-renewal, and the ability to differentiate into all three germ layers in vitro. However, transmission electron microscopy showed a significant increase in the cytoplasm to nucleus ratio and mitochondrial elongation in Dnm1l−/− ESCs, which was due to incomplete fission. To assess the change in metabolic energy, we analyzed oxidative phosphorylation (OXPHOS), glycolysis, and the intracellular ATP concentration. The ESC knockout lines showed an increase in OXPHOS, decrease in glycolysis, and an increase in intracellular ATP concentration, which was related to mitochondrial elongation. In particular, the Dnm1l knockout most significantly affected mitochondrial morphology, energy metabolism, and ATP production in ESCs. Furthermore, RNA sequencing and gene ontology analysis showed that the differentially expressed genes in Mff-/- ESCs were distinct from those in Dnm1l−/- or Fis1−/− ESCs. In total, five metabolism-related genes, namely, Aass, Cdo1, Cyp2b23, Nt5e, and Pck2, were expressed in all three knockout ESC lines, and three of them were associated with regulation of ATP generation.http://www.sciencedirect.com/science/article/pii/S2213231720306340Dynamin 1 like (Dnm1l)Mitochondrial fission factor (Mff)Mitochondrial fission 1 protein (Fis1)KnockoutEmbryonic stem cells (ESCs)Cellular metabolism |
| spellingShingle | Bong Jong Seo Joonhyuk Choi Hyeonwoo La Omer Habib Youngsok Choi Kwonho Hong Jeong Tae Do Role of mitochondrial fission-related genes in mitochondrial morphology and energy metabolism in mouse embryonic stem cells Redox Biology Dynamin 1 like (Dnm1l) Mitochondrial fission factor (Mff) Mitochondrial fission 1 protein (Fis1) Knockout Embryonic stem cells (ESCs) Cellular metabolism |
| title | Role of mitochondrial fission-related genes in mitochondrial morphology and energy metabolism in mouse embryonic stem cells |
| title_full | Role of mitochondrial fission-related genes in mitochondrial morphology and energy metabolism in mouse embryonic stem cells |
| title_fullStr | Role of mitochondrial fission-related genes in mitochondrial morphology and energy metabolism in mouse embryonic stem cells |
| title_full_unstemmed | Role of mitochondrial fission-related genes in mitochondrial morphology and energy metabolism in mouse embryonic stem cells |
| title_short | Role of mitochondrial fission-related genes in mitochondrial morphology and energy metabolism in mouse embryonic stem cells |
| title_sort | role of mitochondrial fission related genes in mitochondrial morphology and energy metabolism in mouse embryonic stem cells |
| topic | Dynamin 1 like (Dnm1l) Mitochondrial fission factor (Mff) Mitochondrial fission 1 protein (Fis1) Knockout Embryonic stem cells (ESCs) Cellular metabolism |
| url | http://www.sciencedirect.com/science/article/pii/S2213231720306340 |
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