Isolation of large dense-core vesicles from bovine adrenal medulla for functional studies
<p dir="ltr">Large dense-core vesicles (LDCVs) contain a variety of neurotransmitters, proteins, and hormones such as biogenic amines and peptides, together with microRNAs (miRNAs). Isolation of LDCVs is essential for functional studies including vesicle fusion, vesicle acidification...
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| مؤلفون آخرون: | , , , |
| منشور في: |
2020
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| _version_ | 1864513519669477376 |
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| author | Yelda Birinci (14152818) |
| author2 | Julia Preobraschenski (14152821) Marcelo Ganzella (11228604) Reinhard Jahn (65883) Yongsoo Park (761850) |
| author2_role | author author author author |
| author_facet | Yelda Birinci (14152818) Julia Preobraschenski (14152821) Marcelo Ganzella (11228604) Reinhard Jahn (65883) Yongsoo Park (761850) |
| author_role | author |
| dc.creator.none.fl_str_mv | Yelda Birinci (14152818) Julia Preobraschenski (14152821) Marcelo Ganzella (11228604) Reinhard Jahn (65883) Yongsoo Park (761850) |
| dc.date.none.fl_str_mv | 2020-05-05T06:00:00Z |
| dc.identifier.none.fl_str_mv | 10.1038/s41598-020-64486-3 |
| dc.relation.none.fl_str_mv | https://figshare.com/articles/journal_contribution/Isolation_of_large_dense-core_vesicles_from_bovine_adrenal_medulla_for_functional_studies/21598167 |
| dc.rights.none.fl_str_mv | CC BY 4.0 info:eu-repo/semantics/openAccess |
| dc.subject.none.fl_str_mv | Biomedical and clinical sciences Medical biochemistry and metabolomics Neurosciences Neurotransmission Vesicle Fusion Large Dense-Core Vesicles (LDCVs) G-Protein-Coupled Receptors (GPCRs) Central Nervous System Peripheral Nervous System Chromaffin Cells Peptides Proteins MicroRNAs (miRNAs) Adrenal Medulla |
| dc.title.none.fl_str_mv | Isolation of large dense-core vesicles from bovine adrenal medulla for functional studies |
| dc.type.none.fl_str_mv | Text Journal contribution info:eu-repo/semantics/publishedVersion text contribution to journal |
| description | <p dir="ltr">Large dense-core vesicles (LDCVs) contain a variety of neurotransmitters, proteins, and hormones such as biogenic amines and peptides, together with microRNAs (miRNAs). Isolation of LDCVs is essential for functional studies including vesicle fusion, vesicle acidification, monoamine transport, and the miRNAs stored in LDCVs. Although several methods were reported for purifying LDCVs, the final fractions are significantly contaminated by other organelles, compromising biochemical characterization. Here we isolated LDCVs (chromaffin granules) with high yield and purity from bovine adrenal medulla. The fractionation protocol combines differential and continuous sucrose gradient centrifugation, allowing for reducing major contaminants such as mitochondria. Purified LDCVs show robust acidification by the endogenous V-ATPase and undergo SNARE-mediated fusion with artificial membranes. Interestingly, LDCVs contain specific miRNAs such as miR-375 and miR-375 is stabilized by protein complex against RNase A. This protocol can be useful in research on the biological functions of LDCVs.</p><p dir="ltr">Author Correction: Isolation of large dense-core vesicles from bovine adrenal medulla for functional studies: <i>Scientific Reports</i> <a href="https://dx.doi.org/10.1038/s41598-022-21361-7" target="_blank">https://dx.doi.org/10.1038/s41598-022-21361-7</a>, published online 06 October 2022.</p><h2>Other Information</h2><p dir="ltr">Published in: Scientific Reports<br>License: <a href="https://creativecommons.org/licenses/by/4.0" target="_blank">https://creativecommons.org/licenses/by/4.0</a><br>See article on publisher's website: <a href="http://dx.doi.org/10.1038/s41598-020-64486-3" target="_blank">http://dx.doi.org/10.1038/s41598-020-64486-3</a></p> |
| eu_rights_str_mv | openAccess |
| id | Manara2_4ffe0e31fe4dce86ee71713751dbaf34 |
| identifier_str_mv | 10.1038/s41598-020-64486-3 |
| network_acronym_str | Manara2 |
| network_name_str | Manara2 |
| oai_identifier_str | oai:figshare.com:article/21598167 |
| publishDate | 2020 |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | CC BY 4.0 |
| spelling | Isolation of large dense-core vesicles from bovine adrenal medulla for functional studiesYelda Birinci (14152818)Julia Preobraschenski (14152821)Marcelo Ganzella (11228604)Reinhard Jahn (65883)Yongsoo Park (761850)Biomedical and clinical sciencesMedical biochemistry and metabolomicsNeurosciencesNeurotransmissionVesicle FusionLarge Dense-Core Vesicles (LDCVs)G-Protein-Coupled Receptors (GPCRs)Central Nervous SystemPeripheral Nervous SystemChromaffin CellsPeptidesProteinsMicroRNAs (miRNAs)Adrenal Medulla<p dir="ltr">Large dense-core vesicles (LDCVs) contain a variety of neurotransmitters, proteins, and hormones such as biogenic amines and peptides, together with microRNAs (miRNAs). Isolation of LDCVs is essential for functional studies including vesicle fusion, vesicle acidification, monoamine transport, and the miRNAs stored in LDCVs. Although several methods were reported for purifying LDCVs, the final fractions are significantly contaminated by other organelles, compromising biochemical characterization. Here we isolated LDCVs (chromaffin granules) with high yield and purity from bovine adrenal medulla. The fractionation protocol combines differential and continuous sucrose gradient centrifugation, allowing for reducing major contaminants such as mitochondria. Purified LDCVs show robust acidification by the endogenous V-ATPase and undergo SNARE-mediated fusion with artificial membranes. Interestingly, LDCVs contain specific miRNAs such as miR-375 and miR-375 is stabilized by protein complex against RNase A. This protocol can be useful in research on the biological functions of LDCVs.</p><p dir="ltr">Author Correction: Isolation of large dense-core vesicles from bovine adrenal medulla for functional studies: <i>Scientific Reports</i> <a href="https://dx.doi.org/10.1038/s41598-022-21361-7" target="_blank">https://dx.doi.org/10.1038/s41598-022-21361-7</a>, published online 06 October 2022.</p><h2>Other Information</h2><p dir="ltr">Published in: Scientific Reports<br>License: <a href="https://creativecommons.org/licenses/by/4.0" target="_blank">https://creativecommons.org/licenses/by/4.0</a><br>See article on publisher's website: <a href="http://dx.doi.org/10.1038/s41598-020-64486-3" target="_blank">http://dx.doi.org/10.1038/s41598-020-64486-3</a></p>2020-05-05T06:00:00ZTextJournal contributioninfo:eu-repo/semantics/publishedVersiontextcontribution to journal10.1038/s41598-020-64486-3https://figshare.com/articles/journal_contribution/Isolation_of_large_dense-core_vesicles_from_bovine_adrenal_medulla_for_functional_studies/21598167CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/215981672020-05-05T06:00:00Z |
| spellingShingle | Isolation of large dense-core vesicles from bovine adrenal medulla for functional studies Yelda Birinci (14152818) Biomedical and clinical sciences Medical biochemistry and metabolomics Neurosciences Neurotransmission Vesicle Fusion Large Dense-Core Vesicles (LDCVs) G-Protein-Coupled Receptors (GPCRs) Central Nervous System Peripheral Nervous System Chromaffin Cells Peptides Proteins MicroRNAs (miRNAs) Adrenal Medulla |
| status_str | publishedVersion |
| title | Isolation of large dense-core vesicles from bovine adrenal medulla for functional studies |
| title_full | Isolation of large dense-core vesicles from bovine adrenal medulla for functional studies |
| title_fullStr | Isolation of large dense-core vesicles from bovine adrenal medulla for functional studies |
| title_full_unstemmed | Isolation of large dense-core vesicles from bovine adrenal medulla for functional studies |
| title_short | Isolation of large dense-core vesicles from bovine adrenal medulla for functional studies |
| title_sort | Isolation of large dense-core vesicles from bovine adrenal medulla for functional studies |
| topic | Biomedical and clinical sciences Medical biochemistry and metabolomics Neurosciences Neurotransmission Vesicle Fusion Large Dense-Core Vesicles (LDCVs) G-Protein-Coupled Receptors (GPCRs) Central Nervous System Peripheral Nervous System Chromaffin Cells Peptides Proteins MicroRNAs (miRNAs) Adrenal Medulla |