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2 Janelia Publications

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    01/10/19 | Cryo-EM of retinoschisin branched networks suggests an intercellular adhesive scaffold in the retina.
    Heymann JB, Vijayasarathy C, Huang RK, Dearborn AD, Sieving PA, Steven AC
    The Journal of Cell Biology. 2019 Jan 10;218(3):1027-38. doi: 10.1083/jcb.201806148

    Mutations in the retinal protein retinoschisin (RS1) cause progressive loss of vision in young males, a form of macular degeneration called X-linked retinoschisis (XLRS). We previously solved the structure of RS1, a 16-mer composed of paired back-to-back octameric rings. Here, we show by cryo-electron microscopy that RS1 16-mers can assemble into extensive branched networks. We classified the different configurations, finding four types of interaction between the RS1 molecules. The predominant configuration is a linear strand with a wavy appearance. Three less frequent types constitute the branch points of the network. In all cases, the "spikes" around the periphery of the double rings are involved in these interactions. In the linear strand, a loop (usually referred to as spike 1) occurs on both sides of the interface between neighboring molecules. Mutations in this loop suppress secretion, indicating the possibility of intracellular higher-order assembly. These observations suggest that branched networks of RS1 may play a stabilizing role in maintaining the integrity of the retina.

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    01/03/19 | High-throughput synapse-resolving two-photon fluorescence microendoscopy for deep-brain volumetric imaging .
    Meng G, Liang Y, Sarsfield S, Jiang W, Lu R, Dudman JT, Aponte Y, Ji N
    eLife. 2019 Jan 03;8:. doi: 10.7554/eLife.40805

    Optical imaging has become a powerful tool for studying brains . The opacity of adult brains makes microendoscopy, with an optical probe such as a gradient index (GRIN) lens embedded into brain tissue to provide optical relay, the method of choice for imaging neurons and neural activity in deeply buried brain structures. Incorporating a Bessel focus scanning module into two-photon fluorescence microendoscopy, we extended the excitation focus axially and improved its lateral resolution. Scanning the Bessel focus in 2D, we imaged volumes of neurons at high-throughput while resolving fine structures such as synaptic terminals. We applied this approach to the volumetric anatomical imaging of dendritic spines and axonal boutons in the mouse hippocampus, and functional imaging of GABAergic neurons in the mouse lateral hypothalamus .

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