Minimal expression of dysferlin prevents development of dysferlinopathy in dysferlin exon 40a knockout mice
Abstract Dysferlin is a Ca2+-activated lipid binding protein implicated in muscle membrane repair. Recessive variants in DYSF result in dysferlinopathy, a progressive muscular dystrophy. We showed previously that calpain cleavage within a motif encoded by alternatively spliced exon 40a releases a 72...
| Main Authors: | , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2023-01-01
|
| Series: | Acta Neuropathologica Communications |
| Online Access: | https://doi.org/10.1186/s40478-022-01473-x |
| _version_ | 1797945675129815040 |
|---|---|
| author | Joe Yasa Claudia E. Reed Adam M. Bournazos Frances J. Evesson Ignatius Pang Mark E. Graham Jesse R. Wark Brunda Nijagal Kim H. Kwan Thomas Kwiatkowski Rachel Jung Noah Weisleder Sandra T. Cooper Frances A. Lemckert |
| author_facet | Joe Yasa Claudia E. Reed Adam M. Bournazos Frances J. Evesson Ignatius Pang Mark E. Graham Jesse R. Wark Brunda Nijagal Kim H. Kwan Thomas Kwiatkowski Rachel Jung Noah Weisleder Sandra T. Cooper Frances A. Lemckert |
| author_sort | Joe Yasa |
| collection | DOAJ |
| description | Abstract Dysferlin is a Ca2+-activated lipid binding protein implicated in muscle membrane repair. Recessive variants in DYSF result in dysferlinopathy, a progressive muscular dystrophy. We showed previously that calpain cleavage within a motif encoded by alternatively spliced exon 40a releases a 72 kDa C-terminal minidysferlin recruited to injured sarcolemma. Herein we use CRISPR/Cas9 gene editing to knock out murine Dysf exon 40a, to specifically assess its role in membrane repair and development of dysferlinopathy. We created three Dysf exon 40a knockout (40aKO) mouse lines that each express different levels of dysferlin protein ranging from ~ 90%, ~ 50% and ~ 10–20% levels of wild-type. Histopathological analysis of skeletal muscles from all 12-month-old 40aKO lines showed virtual absence of dystrophic features and normal membrane repair capacity for all three 40aKO lines, as compared with dysferlin-null BLAJ mice. Further, lipidomic and proteomic analyses on 18wk old quadriceps show all three 40aKO lines are spared the profound lipidomic/proteomic imbalance that characterises dysferlin-deficient BLAJ muscles. Collective results indicate that membrane repair does not depend upon calpain cleavage within exon 40a and that ~ 10–20% of WT dysferlin protein expression is sufficient to maintain the muscle lipidome, proteome and membrane repair capacity to crucially prevent development of dysferlinopathy. |
| first_indexed | 2024-04-10T20:58:51Z |
| format | Article |
| id | doaj.art-6b549e68706942818cd58183122bbf7a |
| institution | Directory Open Access Journal |
| issn | 2051-5960 |
| language | English |
| last_indexed | 2024-04-10T20:58:51Z |
| publishDate | 2023-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Acta Neuropathologica Communications |
| spelling | doaj.art-6b549e68706942818cd58183122bbf7a2023-01-22T12:28:15ZengBMCActa Neuropathologica Communications2051-59602023-01-0111111610.1186/s40478-022-01473-xMinimal expression of dysferlin prevents development of dysferlinopathy in dysferlin exon 40a knockout miceJoe Yasa0Claudia E. Reed1Adam M. Bournazos2Frances J. Evesson3Ignatius Pang4Mark E. Graham5Jesse R. Wark6Brunda Nijagal7Kim H. Kwan8Thomas Kwiatkowski9Rachel Jung10Noah Weisleder11Sandra T. Cooper12Frances A. Lemckert13Kids Neuroscience Centre, The Children’s Hospital at WestmeadKids Neuroscience Centre, The Children’s Hospital at WestmeadKids Neuroscience Centre, The Children’s Hospital at WestmeadKids Neuroscience Centre, The Children’s Hospital at WestmeadSynapse Proteomics, Children’s Medical Research Institute, The University of SydneySynapse Proteomics, Children’s Medical Research Institute, The University of SydneyOperations, Children’s Medical Research Institute, The University of SydneyMetabolomics Australia, Bio21 Institute, The University of MelbourneMetabolomics Australia, Bio21 Institute, The University of MelbourneWest Chester UniversityDepartment of Physiology and Cell Biology, Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University Wexner Medical CenterDepartment of Physiology and Cell Biology, Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University Wexner Medical CenterKids Neuroscience Centre, The Children’s Hospital at WestmeadKids Neuroscience Centre, The Children’s Hospital at WestmeadAbstract Dysferlin is a Ca2+-activated lipid binding protein implicated in muscle membrane repair. Recessive variants in DYSF result in dysferlinopathy, a progressive muscular dystrophy. We showed previously that calpain cleavage within a motif encoded by alternatively spliced exon 40a releases a 72 kDa C-terminal minidysferlin recruited to injured sarcolemma. Herein we use CRISPR/Cas9 gene editing to knock out murine Dysf exon 40a, to specifically assess its role in membrane repair and development of dysferlinopathy. We created three Dysf exon 40a knockout (40aKO) mouse lines that each express different levels of dysferlin protein ranging from ~ 90%, ~ 50% and ~ 10–20% levels of wild-type. Histopathological analysis of skeletal muscles from all 12-month-old 40aKO lines showed virtual absence of dystrophic features and normal membrane repair capacity for all three 40aKO lines, as compared with dysferlin-null BLAJ mice. Further, lipidomic and proteomic analyses on 18wk old quadriceps show all three 40aKO lines are spared the profound lipidomic/proteomic imbalance that characterises dysferlin-deficient BLAJ muscles. Collective results indicate that membrane repair does not depend upon calpain cleavage within exon 40a and that ~ 10–20% of WT dysferlin protein expression is sufficient to maintain the muscle lipidome, proteome and membrane repair capacity to crucially prevent development of dysferlinopathy.https://doi.org/10.1186/s40478-022-01473-x |
| spellingShingle | Joe Yasa Claudia E. Reed Adam M. Bournazos Frances J. Evesson Ignatius Pang Mark E. Graham Jesse R. Wark Brunda Nijagal Kim H. Kwan Thomas Kwiatkowski Rachel Jung Noah Weisleder Sandra T. Cooper Frances A. Lemckert Minimal expression of dysferlin prevents development of dysferlinopathy in dysferlin exon 40a knockout mice Acta Neuropathologica Communications |
| title | Minimal expression of dysferlin prevents development of dysferlinopathy in dysferlin exon 40a knockout mice |
| title_full | Minimal expression of dysferlin prevents development of dysferlinopathy in dysferlin exon 40a knockout mice |
| title_fullStr | Minimal expression of dysferlin prevents development of dysferlinopathy in dysferlin exon 40a knockout mice |
| title_full_unstemmed | Minimal expression of dysferlin prevents development of dysferlinopathy in dysferlin exon 40a knockout mice |
| title_short | Minimal expression of dysferlin prevents development of dysferlinopathy in dysferlin exon 40a knockout mice |
| title_sort | minimal expression of dysferlin prevents development of dysferlinopathy in dysferlin exon 40a knockout mice |
| url | https://doi.org/10.1186/s40478-022-01473-x |
| work_keys_str_mv | AT joeyasa minimalexpressionofdysferlinpreventsdevelopmentofdysferlinopathyindysferlinexon40aknockoutmice AT claudiaereed minimalexpressionofdysferlinpreventsdevelopmentofdysferlinopathyindysferlinexon40aknockoutmice AT adammbournazos minimalexpressionofdysferlinpreventsdevelopmentofdysferlinopathyindysferlinexon40aknockoutmice AT francesjevesson minimalexpressionofdysferlinpreventsdevelopmentofdysferlinopathyindysferlinexon40aknockoutmice AT ignatiuspang minimalexpressionofdysferlinpreventsdevelopmentofdysferlinopathyindysferlinexon40aknockoutmice AT markegraham minimalexpressionofdysferlinpreventsdevelopmentofdysferlinopathyindysferlinexon40aknockoutmice AT jesserwark minimalexpressionofdysferlinpreventsdevelopmentofdysferlinopathyindysferlinexon40aknockoutmice AT brundanijagal minimalexpressionofdysferlinpreventsdevelopmentofdysferlinopathyindysferlinexon40aknockoutmice AT kimhkwan minimalexpressionofdysferlinpreventsdevelopmentofdysferlinopathyindysferlinexon40aknockoutmice AT thomaskwiatkowski minimalexpressionofdysferlinpreventsdevelopmentofdysferlinopathyindysferlinexon40aknockoutmice AT racheljung minimalexpressionofdysferlinpreventsdevelopmentofdysferlinopathyindysferlinexon40aknockoutmice AT noahweisleder minimalexpressionofdysferlinpreventsdevelopmentofdysferlinopathyindysferlinexon40aknockoutmice AT sandratcooper minimalexpressionofdysferlinpreventsdevelopmentofdysferlinopathyindysferlinexon40aknockoutmice AT francesalemckert minimalexpressionofdysferlinpreventsdevelopmentofdysferlinopathyindysferlinexon40aknockoutmice |