Generation of interconnected vesicles in a liposomal cell model
| Autor: | Daniel Fjällborg, Tatsiana Lobovkina, Kiryl Kustanovich, Baharan Ali Doosti, Ann-Sofie Cans, Aldo Jesorka |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Microinjections Biophysics Phospholipid lcsh:Medicine Endocytosis Physical Chemistry Models Biological Article Imaging studies 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Biophysical chemistry Other Basic Medicine lcsh:Science Microinjection Phospholipids Synthetic biology Unilamellar Liposomes Neurotransmitter Agents Liposome Membranes Nanotubes Multidisciplinary Vesicle lcsh:R Bioinspired materials Pipette Secretory Vesicle 030104 developmental biology Membrane Microscopy Fluorescence chemistry lcsh:Q Calcium Chemical tools 030217 neurology & neurosurgery |
| Zdroj: | Scientific Reports Scientific Reports, Vol 10, Iss 1, Pp 1-7 (2020) Scientific Reports (2045-2322) vol.10(2020) |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/s41598-020-70562-5 |
| Popis: | We introduce an experimental method based upon a glass micropipette microinjection technique for generating a multitude of interconnected vesicles (IVs) in the interior of a single giant unilamellar phospholipid vesicle (GUV) serving as a cell model system. The GUV membrane, consisting of a mixture of soybean polar lipid extract and anionic phosphatidylserine, is adhered to a multilamellar lipid vesicle that functions as a lipid reservoir. Continuous IV formation was achieved by bringing a micropipette in direct contact with the outer GUV surface and subjecting it to a localized stream of a Ca2+ solution from the micropipette tip. IVs are rapidly and sequentially generated and inserted into the GUV interior and encapsulate portions of the micropipette fluid content. The IVs remain connected to the GUV membrane and are interlinked by short lipid nanotubes and resemble beads on a string. The vesicle chain-growth from the GUV membrane is maintained for as long as there is the supply of membrane material and Ca2+ solution, and the size of the individual IVs is controlled by the diameter of the micropipette tip. We also demonstrate that the IVs can be co-loaded with high concentrations of neurotransmitter and protein molecules and displaying a steep calcium ion concentration gradient across the membrane. These characteristics are analogous to native secretory vesicles and could, therefore, serve as a model system for studying secretory mechanisms in biological systems. |
| Databáze: | OpenAIRE |
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