Novel Isolation Method Reveals Sex-Specific Composition and Neurotoxicity of Small Extracellular Vesicles in a Mouse Model of Alzheimer’s Disease
We developed a new method to isolate small extracellular vesicles (sEVs) from male and female wild-type and 5xFAD mouse brains to investigate the sex-specific functions of sEVs in Alzheimer’s disease (AD). A mass spectrometric analysis revealed that sEVs contained proteins critical for EV formation...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-06-01
|
| Series: | Cells |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2073-4409/12/12/1623 |
| _version_ | 1797595588307451904 |
|---|---|
| author | Ahmed Elsherbini Zhihui Zhu Zainuddin Quadri Simone M. Crivelli Xiaojia Ren Hemendra J. Vekaria Priyanka Tripathi Liping Zhang Wenbo Zhi Erhard Bieberich |
| author_facet | Ahmed Elsherbini Zhihui Zhu Zainuddin Quadri Simone M. Crivelli Xiaojia Ren Hemendra J. Vekaria Priyanka Tripathi Liping Zhang Wenbo Zhi Erhard Bieberich |
| author_sort | Ahmed Elsherbini |
| collection | DOAJ |
| description | We developed a new method to isolate small extracellular vesicles (sEVs) from male and female wild-type and 5xFAD mouse brains to investigate the sex-specific functions of sEVs in Alzheimer’s disease (AD). A mass spectrometric analysis revealed that sEVs contained proteins critical for EV formation and Aβ. ExoView analysis showed that female mice contained more GFAP and Aβ-labeled sEVs, suggesting that a larger proportion of sEVs from the female brain is derived from astrocytes and/or more likely to bind to Aβ. Moreover, sEVs from female brains had more acid sphingomyelinase (ASM) and ceramide, an enzyme and its sphingolipid product important for EV formation and Aβ binding to EVs, respectively. We confirmed the function of ASM in EV formation and Aβ binding using co-labeling and proximity ligation assays, showing that ASM inhibitors prevented complex formation between Aβ and ceramide in primary cultured astrocytes. Finally, our study demonstrated that sEVs from female 5xFAD mice were more neurotoxic than those from males, as determined by impaired mitochondrial function (Seahorse assays) and LDH cytotoxicity assays. Our study suggests that sex-specific sEVs are functionally distinct markers for AD and that ASM is a potential target for AD therapy. |
| first_indexed | 2024-03-11T02:38:43Z |
| format | Article |
| id | doaj.art-4c6ee4e2a1484f0dab57d7d2b918123a |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-11T02:38:43Z |
| publishDate | 2023-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-4c6ee4e2a1484f0dab57d7d2b918123a2023-11-18T09:43:35ZengMDPI AGCells2073-44092023-06-011212162310.3390/cells12121623Novel Isolation Method Reveals Sex-Specific Composition and Neurotoxicity of Small Extracellular Vesicles in a Mouse Model of Alzheimer’s DiseaseAhmed Elsherbini0Zhihui Zhu1Zainuddin Quadri2Simone M. Crivelli3Xiaojia Ren4Hemendra J. Vekaria5Priyanka Tripathi6Liping Zhang7Wenbo Zhi8Erhard Bieberich9Department of Physiology, University of Kentucky College of Medicine, Lexington, KY 40536, USADepartment of Physiology, University of Kentucky College of Medicine, Lexington, KY 40536, USADepartment of Physiology, University of Kentucky College of Medicine, Lexington, KY 40536, USADepartment of Physiology, University of Kentucky College of Medicine, Lexington, KY 40536, USADepartment of Physiology, University of Kentucky College of Medicine, Lexington, KY 40536, USASpinal Cord and Brain Injury Research Center (SCoBIRC), University of Kentucky, Lexington, KY 40536, USADepartment of Physiology, University of Kentucky College of Medicine, Lexington, KY 40536, USADepartment of Physiology, University of Kentucky College of Medicine, Lexington, KY 40536, USADepartment of Center for Biotechnology and Genomic Medicine, Augusta University, Augusta, GA 30912, USADepartment of Physiology, University of Kentucky College of Medicine, Lexington, KY 40536, USAWe developed a new method to isolate small extracellular vesicles (sEVs) from male and female wild-type and 5xFAD mouse brains to investigate the sex-specific functions of sEVs in Alzheimer’s disease (AD). A mass spectrometric analysis revealed that sEVs contained proteins critical for EV formation and Aβ. ExoView analysis showed that female mice contained more GFAP and Aβ-labeled sEVs, suggesting that a larger proportion of sEVs from the female brain is derived from astrocytes and/or more likely to bind to Aβ. Moreover, sEVs from female brains had more acid sphingomyelinase (ASM) and ceramide, an enzyme and its sphingolipid product important for EV formation and Aβ binding to EVs, respectively. We confirmed the function of ASM in EV formation and Aβ binding using co-labeling and proximity ligation assays, showing that ASM inhibitors prevented complex formation between Aβ and ceramide in primary cultured astrocytes. Finally, our study demonstrated that sEVs from female 5xFAD mice were more neurotoxic than those from males, as determined by impaired mitochondrial function (Seahorse assays) and LDH cytotoxicity assays. Our study suggests that sex-specific sEVs are functionally distinct markers for AD and that ASM is a potential target for AD therapy.https://www.mdpi.com/2073-4409/12/12/1623extracellular vesiclesAlzheimer’sacid sphingomyelinaseceramideneurotoxicity |
| spellingShingle | Ahmed Elsherbini Zhihui Zhu Zainuddin Quadri Simone M. Crivelli Xiaojia Ren Hemendra J. Vekaria Priyanka Tripathi Liping Zhang Wenbo Zhi Erhard Bieberich Novel Isolation Method Reveals Sex-Specific Composition and Neurotoxicity of Small Extracellular Vesicles in a Mouse Model of Alzheimer’s Disease Cells extracellular vesicles Alzheimer’s acid sphingomyelinase ceramide neurotoxicity |
| title | Novel Isolation Method Reveals Sex-Specific Composition and Neurotoxicity of Small Extracellular Vesicles in a Mouse Model of Alzheimer’s Disease |
| title_full | Novel Isolation Method Reveals Sex-Specific Composition and Neurotoxicity of Small Extracellular Vesicles in a Mouse Model of Alzheimer’s Disease |
| title_fullStr | Novel Isolation Method Reveals Sex-Specific Composition and Neurotoxicity of Small Extracellular Vesicles in a Mouse Model of Alzheimer’s Disease |
| title_full_unstemmed | Novel Isolation Method Reveals Sex-Specific Composition and Neurotoxicity of Small Extracellular Vesicles in a Mouse Model of Alzheimer’s Disease |
| title_short | Novel Isolation Method Reveals Sex-Specific Composition and Neurotoxicity of Small Extracellular Vesicles in a Mouse Model of Alzheimer’s Disease |
| title_sort | novel isolation method reveals sex specific composition and neurotoxicity of small extracellular vesicles in a mouse model of alzheimer s disease |
| topic | extracellular vesicles Alzheimer’s acid sphingomyelinase ceramide neurotoxicity |
| url | https://www.mdpi.com/2073-4409/12/12/1623 |
| work_keys_str_mv | AT ahmedelsherbini novelisolationmethodrevealssexspecificcompositionandneurotoxicityofsmallextracellularvesiclesinamousemodelofalzheimersdisease AT zhihuizhu novelisolationmethodrevealssexspecificcompositionandneurotoxicityofsmallextracellularvesiclesinamousemodelofalzheimersdisease AT zainuddinquadri novelisolationmethodrevealssexspecificcompositionandneurotoxicityofsmallextracellularvesiclesinamousemodelofalzheimersdisease AT simonemcrivelli novelisolationmethodrevealssexspecificcompositionandneurotoxicityofsmallextracellularvesiclesinamousemodelofalzheimersdisease AT xiaojiaren novelisolationmethodrevealssexspecificcompositionandneurotoxicityofsmallextracellularvesiclesinamousemodelofalzheimersdisease AT hemendrajvekaria novelisolationmethodrevealssexspecificcompositionandneurotoxicityofsmallextracellularvesiclesinamousemodelofalzheimersdisease AT priyankatripathi novelisolationmethodrevealssexspecificcompositionandneurotoxicityofsmallextracellularvesiclesinamousemodelofalzheimersdisease AT lipingzhang novelisolationmethodrevealssexspecificcompositionandneurotoxicityofsmallextracellularvesiclesinamousemodelofalzheimersdisease AT wenbozhi novelisolationmethodrevealssexspecificcompositionandneurotoxicityofsmallextracellularvesiclesinamousemodelofalzheimersdisease AT erhardbieberich novelisolationmethodrevealssexspecificcompositionandneurotoxicityofsmallextracellularvesiclesinamousemodelofalzheimersdisease |