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b PH G fluorescence (F PH, normalized to baseline), A532 fluorescence (F 532, normalized to baseline), and STED XZ/Y fix images (at times indicated with lines) showing Flat→Λ→Ω→O transition induced by depol 1s (gray triangle). Lower: sampled ICa and Cm induced by depol 1s. Government work and not under copyright protection in the US foreign copyright protection may apply.Ī Upper: setup drawing – a PH G-labeled cell (green) in A532-containing bath (red) with whole-cell recording of calcium currents (ICa) and capacitance (Cm). Their impact is widespread beyond secretory cells, as the unexpectedly powerful functions of dynamin and actin, previously thought to mediate fission and overcome tension, respectively, may contribute to many dynamin/actin-dependent non-coated-membrane buddings, coated-membrane buddings, and other membrane remodeling processes. These mechanisms control budding speed, vesicle size and number, generating diverse endocytic modes differing in these parameters. By visualizing proteins in mediating endocytic budding in live neuroendocrine cells, performing in vitro protein reconstitution and physical modeling, we discovered how non-coated-membrane budding is mediated: actin filaments and dynamin generate a pulling force transforming flat membrane into Λ-shape subsequently, dynamin helices surround and constrict Λ-profile's base, transforming Λ- to Ω-profile, and then constrict Ω-profile's pore, converting Ω-profiles to vesicles.
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However, forces mediating many non-coated membrane buddings remain unclear. Accumulated studies have identified coat-proteins (e.g., clathrin) as potential budding factors. Membrane budding entails forces to transform flat membrane into vesicles essential for cell survival.