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2 Janelia Publications

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    11/22/21 | De novo endocytic clathrin coats develop curvature at early stages of their formation.
    Willy NM, Ferguson JP, Akatay A, Huber S, Djakbarova U, Silahli S, Cakez C, Hasan F, Chang HC, Travesset A, Li S, Zandi R, Li D, Betzig E, Cocucci E, Kural C
    Development Cell. 2021 Nov 22;56(22):3146-3159.e5. doi: 10.1016/j.devcel.2021.10.019

    Sculpting a flat patch of membrane into an endocytic vesicle requires curvature generation on the cell surface, which is the primary function of the endocytosis machinery. Using super-resolved live cell fluorescence imaging, we demonstrate that curvature generation by individual clathrin-coated pits can be detected in real time within cultured cells and tissues of developing organisms. Our analyses demonstrate that the footprint of clathrin coats increases monotonically during the formation of pits at different levels of plasma membrane tension. These findings are only compatible with models that predict curvature generation at the early stages of endocytic clathrin pit formation. We also found that CALM adaptors associated with clathrin plaques form clusters, whereas AP2 distribution is more homogenous. Considering the curvature sensing and driving roles of CALM, we propose that CALM clusters may increase the strain on clathrin lattices locally, eventually giving rise to rupture and subsequent pit completion at the edges of plaques.

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    09/30/21 | Cellular bases of olfactory circuit assembly revealed by systematic time-lapse imaging.
    Li T, Fu T, Wong KK, Li H, Xie Q, Luginbuhl DJ, Wagner MJ, Betzig E, Luo L
    Cell. 2021 Sep 30;184(20):5107. doi: 10.1016/j.cell.2021.08.030

    Neural circuit assembly features simultaneous targeting of numerous neuronal processes from constituent neuron types, yet the dynamics is poorly understood. Here, we use the Drosophila olfactory circuit to investigate dynamic cellular processes by which olfactory receptor neurons (ORNs) target axons precisely to specific glomeruli in the ipsi- and contralateral antennal lobes. Time-lapse imaging of individual axons from 30 ORN types revealed a rich diversity in extension speed, innervation timing, and ipsilateral branch locations and identified that ipsilateral targeting occurs via stabilization of transient interstitial branches. Fast imaging using adaptive optics-corrected lattice light-sheet microscopy showed that upon approaching target, many ORN types exhibiting "exploring branches" consisted of parallel microtubule-based terminal branches emanating from an F-actin-rich hub. Antennal nerve ablations uncovered essential roles for bilateral axons in contralateral target selection and for ORN axons to facilitate dendritic refinement of postsynaptic partner neurons. Altogether, these observations provide cellular bases for wiring specificity establishment.

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