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Draft, Ryan

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Draft, Ryan

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2013 - 20152

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Author's Publications: search.filters.isAuthorOfPublication.5da19dec-d311-42e2-980f-520f90a5d0bc

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    Developmental Bias in Cleavage-Stage Mouse Blastomeres
    (Elsevier BV, 2013) Tabansky, Inna; Lenarcic, Alan; Draft, Ryan; Loulier, Karine; Keskin, Derin Benerci; Rosains, Jacqueline; Rivera-Feliciano, Jose; Lichtman, Jeff; Livet, Jean; Stern, Joel N H; Sanes, Joshua; Eggan, Kevin
    BACKGROUND: The cleavage-stage mouse embryo is composed of superficially equivalent blastomeres that will generate both the embryonic inner cell mass (ICM) and the supportive trophectoderm (TE). However, it remains unsettled whether the contribution of each blastomere to these two lineages can be accounted for by chance. Addressing the question of blastomere cell fate may be of practical importance, because preimplantation genetic diagnosis requires removal of blastomeres from the early human embryo. To determine whether blastomere allocation to the two earliest lineages is random, we developed and utilized a recombination-mediated, noninvasive combinatorial fluorescent labeling method for embryonic lineage tracing. RESULTS: When we induced recombination at cleavage stages, we observed a statistically significant bias in the contribution of the resulting labeled clones to the trophectoderm or the inner cell mass in a subset of embryos. Surprisingly, we did not find a correlation between localization of clones in the embryonic and abembryonic hemispheres of the late blastocyst and their allocation to the TE and ICM, suggesting that TE-ICM bias arises separately from embryonic-abembryonic bias. Rainbow lineage tracing also allowed us to demonstrate that the bias observed in the blastocyst persists into postimplantation stages and therefore has relevance for subsequent development. CONCLUSIONS: The Rainbow transgenic mice that we describe here have allowed us to detect lineage-dependent bias in early development. They should also enable assessment of the developmental equivalence of mammalian progenitor cells in a variety of tissues.
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    Multispectral labeling technique to map many neighboring axonal projections in the same tissue
    (Nature Publishing Group, 2015) Tsuriel, Shlomo; Gudes, Sagi; Draft, Ryan; Binshtok, Alexander M.; Lichtman, Jeff
    We describe a method to map the location of axonal arbors of many individual neurons simultaneously based on the spectral properties of retrogradely transported dyelabeled vesicles. We inject overlapping regions of an axon target area with three or more different colored retrograde tracers. Based on the combinations and intensities of the colors in the individual vesicles transported to neuronal somata we calculate the projection sites of each neuron’s axon.This neuronal positioning system (NPS) enables mapping of many axons in a simple automated way. NPS combined with spectral (Brainbow) labeling of the input to autonomic ganglion cells show that the locations of ganglion cell projections to a mouse salivary gland relate to the identities of their preganglionic axonal innervation. We also show that NPS can delineate projections of many axons simultaneously in the mouse CNS.