Specific induction of TUNA during neural differentiation of mESCs. (A) Gene expression during in vitro ESC differentiation towards the mesodermal lineage. mESC cells and RNA were harvested on the indicated days. Expression of KDR, Tal1 (mesoderm lineage markers), TUNA, Nestin, and Oct4 were measured by qRT-PCR. Transcript levels were normalized to Actb and compared with expression on day 0. Data are the mean ± SD of triplicates. (B) Behavioral response of zebrafish larvae to a touch stimulus. Embryos were microinjected with 1 ng of a mixture of tuna MOs or a control MO, and a touch response test was performed at 3 dpf. Images are single frames from a movie showing the locomotory response of zebrafish injected with tuna MOs (ii and ii2), compared with animals injected with the control MO (i and i2). (C) Quantification of zebrafish larvae showing impaired locomotor function. Embryos were microinjected with 1ng of three individual tuna MOs, mixed MOs, or a control MO. Touch response test was performed at 3 dpf. The percentage of embryos demonstrating behavioral defects is shown (n H 50). (D) Animals showed no apparent skeletal muscle defects upon tuna MO treatment, indicating that the behavioral phenotype is a reflection of tuna MO effects on the CNS.
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