Protocol for measuring force in skinned diaphragm muscle fibers of myopathic SEPN1 knockout mice following chronic tauroursodeoxycholic acid treatment
Summary: Selenoprotein N1 (SEPN1) is a type II endoplasmic reticulum (ER) glycoprotein. Loss-of-function mutations in the gene encoding for SEPN1 give rise to myopathy. Here, we present a protocol for evaluating the contractility of diaphragmatic muscle fibers of SEPN1 knockout mice following chroni...
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| Language: | English |
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Elsevier
2025-09-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725003247 |
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| author | Leonardo Nogara Serena Germani Cosimo De Napoli Bert Blaauw Ester Zito |
| author_facet | Leonardo Nogara Serena Germani Cosimo De Napoli Bert Blaauw Ester Zito |
| author_sort | Leonardo Nogara |
| collection | DOAJ |
| description | Summary: Selenoprotein N1 (SEPN1) is a type II endoplasmic reticulum (ER) glycoprotein. Loss-of-function mutations in the gene encoding for SEPN1 give rise to myopathy. Here, we present a protocol for evaluating the contractility of diaphragmatic muscle fibers of SEPN1 knockout mice following chronic treatment with tauroursodeoxycholic acid (TUDCA). We describe steps for genotyping SEPN1 knockout mice, TUDCA in vivo treatment, diaphragm dissection, and chemical permeabilization. We then detail procedures for single muscle fiber isolation and tension measurement.For complete details on the use and execution of this protocol, please refer to Germani et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-eb75f636a34a4e4ab7ada181f7570355 |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-eb75f636a34a4e4ab7ada181f75703552025-08-20T03:32:12ZengElsevierSTAR Protocols2666-16672025-09-016310391810.1016/j.xpro.2025.103918Protocol for measuring force in skinned diaphragm muscle fibers of myopathic SEPN1 knockout mice following chronic tauroursodeoxycholic acid treatmentLeonardo Nogara0Serena Germani1Cosimo De Napoli2Bert Blaauw3Ester Zito4Department of Biomedical Sciences, University of Padua, Padua, Italy; Department of Pharmaceutical Sciences, University of Padua, Padua, Italy; Corresponding authorIstituto di Ricerche Farmacologiche Mario Negri IRCCS, Milan, Italy; Corresponding authorDepartment of Biomedical Sciences, University of Padua, Padua, ItalyDepartment of Biomedical Sciences, University of Padua, Padua, Italy; Veneto Institute of Molecular Medicine (VIMM), Padua, ItalyIstituto di Ricerche Farmacologiche Mario Negri IRCCS, Milan, Italy; Department of Biomolecular Sciences, University of Urbino Carlo Bo, Urbino, Italy; Corresponding authorSummary: Selenoprotein N1 (SEPN1) is a type II endoplasmic reticulum (ER) glycoprotein. Loss-of-function mutations in the gene encoding for SEPN1 give rise to myopathy. Here, we present a protocol for evaluating the contractility of diaphragmatic muscle fibers of SEPN1 knockout mice following chronic treatment with tauroursodeoxycholic acid (TUDCA). We describe steps for genotyping SEPN1 knockout mice, TUDCA in vivo treatment, diaphragm dissection, and chemical permeabilization. We then detail procedures for single muscle fiber isolation and tension measurement.For complete details on the use and execution of this protocol, please refer to Germani et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166725003247Biotechnology and bioengineeringHealth SciencesModel Organisms |
| spellingShingle | Leonardo Nogara Serena Germani Cosimo De Napoli Bert Blaauw Ester Zito Protocol for measuring force in skinned diaphragm muscle fibers of myopathic SEPN1 knockout mice following chronic tauroursodeoxycholic acid treatment STAR Protocols Biotechnology and bioengineering Health Sciences Model Organisms |
| title | Protocol for measuring force in skinned diaphragm muscle fibers of myopathic SEPN1 knockout mice following chronic tauroursodeoxycholic acid treatment |
| title_full | Protocol for measuring force in skinned diaphragm muscle fibers of myopathic SEPN1 knockout mice following chronic tauroursodeoxycholic acid treatment |
| title_fullStr | Protocol for measuring force in skinned diaphragm muscle fibers of myopathic SEPN1 knockout mice following chronic tauroursodeoxycholic acid treatment |
| title_full_unstemmed | Protocol for measuring force in skinned diaphragm muscle fibers of myopathic SEPN1 knockout mice following chronic tauroursodeoxycholic acid treatment |
| title_short | Protocol for measuring force in skinned diaphragm muscle fibers of myopathic SEPN1 knockout mice following chronic tauroursodeoxycholic acid treatment |
| title_sort | protocol for measuring force in skinned diaphragm muscle fibers of myopathic sepn1 knockout mice following chronic tauroursodeoxycholic acid treatment |
| topic | Biotechnology and bioengineering Health Sciences Model Organisms |
| url | http://www.sciencedirect.com/science/article/pii/S2666166725003247 |
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