Generation of a Transgenic Mouse Model for Investigating Mitochondria in Sperm
Mitochondria play a crucial role in sperm development; however, the mechanisms regulating their function in sperm remain poorly understood. Developing a method to regulate the expression of a target gene within the mitochondria of sperm is a vital step in this area of research. In this study, we aim...
Saved in:
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2025-02-01
|
| Series: | Cells |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2073-4409/14/4/296 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849719053257539584 |
|---|---|
| author | Hironmoy Sarkar Suryaprakash R. Batta Neerja Wadhwa Subeer S. Majumdar Bhola Shankar Pradhan |
| author_facet | Hironmoy Sarkar Suryaprakash R. Batta Neerja Wadhwa Subeer S. Majumdar Bhola Shankar Pradhan |
| author_sort | Hironmoy Sarkar |
| collection | DOAJ |
| description | Mitochondria play a crucial role in sperm development; however, the mechanisms regulating their function in sperm remain poorly understood. Developing a method to regulate the expression of a target gene within the mitochondria of sperm is a vital step in this area of research. In this study, we aimed to create a system for expressing a transgene in the mitochondria of sperm. As a proof of concept, we generated transgenic mice that express green fluorescent protein (GFP) fused with a mitochondrial localization signal (MLS) driven by the phosphoglycerate kinase 2 (PGK2) promoter, which facilitates the transgene expression in the sperm. Although the PGK2 promoter has previously shown to drive gene expression in spermatocytes and spermatids, the novelty of our approach lies in the combination of PGK2-driven MLS-GFP expression to study mitochondria in vivo. We established two founder lines of transgenic mice through pronuclear microinjection, and MLS-GFP expression was confirmed in the mitochondria of sperm cells using fluorescence microscopy and flow cytometry. Consequently, we provide a novel platform for investigating mitochondrial function in sperm, where GFP can be substituted with other genes of interest to examine their effects on mitochondria. This system specifically targets sperm mitochondria, offering an innovative approach for studying mitochondrial function in vivo. |
| format | Article |
| id | doaj-art-6f1d170130954d69974da7fa396d8d5c |
| institution | DOAJ |
| issn | 2073-4409 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj-art-6f1d170130954d69974da7fa396d8d5c2025-08-20T03:12:14ZengMDPI AGCells2073-44092025-02-0114429610.3390/cells14040296Generation of a Transgenic Mouse Model for Investigating Mitochondria in SpermHironmoy Sarkar0Suryaprakash R. Batta1Neerja Wadhwa2Subeer S. Majumdar3Bhola Shankar Pradhan4Cellular Endocrinology Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, JNU Complex, New Delhi 110067, IndiaCellular Endocrinology Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, JNU Complex, New Delhi 110067, IndiaEmbryo Biotechnology Lab, National Institute of Immunology, New Delhi 110067, IndiaCellular Endocrinology Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, JNU Complex, New Delhi 110067, IndiaCellular Endocrinology Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, JNU Complex, New Delhi 110067, IndiaMitochondria play a crucial role in sperm development; however, the mechanisms regulating their function in sperm remain poorly understood. Developing a method to regulate the expression of a target gene within the mitochondria of sperm is a vital step in this area of research. In this study, we aimed to create a system for expressing a transgene in the mitochondria of sperm. As a proof of concept, we generated transgenic mice that express green fluorescent protein (GFP) fused with a mitochondrial localization signal (MLS) driven by the phosphoglycerate kinase 2 (PGK2) promoter, which facilitates the transgene expression in the sperm. Although the PGK2 promoter has previously shown to drive gene expression in spermatocytes and spermatids, the novelty of our approach lies in the combination of PGK2-driven MLS-GFP expression to study mitochondria in vivo. We established two founder lines of transgenic mice through pronuclear microinjection, and MLS-GFP expression was confirmed in the mitochondria of sperm cells using fluorescence microscopy and flow cytometry. Consequently, we provide a novel platform for investigating mitochondrial function in sperm, where GFP can be substituted with other genes of interest to examine their effects on mitochondria. This system specifically targets sperm mitochondria, offering an innovative approach for studying mitochondrial function in vivo.https://www.mdpi.com/2073-4409/14/4/296mitochondriaPGK2MLSspermtransgenic mice |
| spellingShingle | Hironmoy Sarkar Suryaprakash R. Batta Neerja Wadhwa Subeer S. Majumdar Bhola Shankar Pradhan Generation of a Transgenic Mouse Model for Investigating Mitochondria in Sperm Cells mitochondria PGK2 MLS sperm transgenic mice |
| title | Generation of a Transgenic Mouse Model for Investigating Mitochondria in Sperm |
| title_full | Generation of a Transgenic Mouse Model for Investigating Mitochondria in Sperm |
| title_fullStr | Generation of a Transgenic Mouse Model for Investigating Mitochondria in Sperm |
| title_full_unstemmed | Generation of a Transgenic Mouse Model for Investigating Mitochondria in Sperm |
| title_short | Generation of a Transgenic Mouse Model for Investigating Mitochondria in Sperm |
| title_sort | generation of a transgenic mouse model for investigating mitochondria in sperm |
| topic | mitochondria PGK2 MLS sperm transgenic mice |
| url | https://www.mdpi.com/2073-4409/14/4/296 |
| work_keys_str_mv | AT hironmoysarkar generationofatransgenicmousemodelforinvestigatingmitochondriainsperm AT suryaprakashrbatta generationofatransgenicmousemodelforinvestigatingmitochondriainsperm AT neerjawadhwa generationofatransgenicmousemodelforinvestigatingmitochondriainsperm AT subeersmajumdar generationofatransgenicmousemodelforinvestigatingmitochondriainsperm AT bholashankarpradhan generationofatransgenicmousemodelforinvestigatingmitochondriainsperm |