Germ-line knockouts of mbd5 are adult viable and suffer from behavioral abnormalities. (A) Predicted amino acid sequences of wild type and mbd5 mutant alleles. (B–D) Western-blot shows reduced Mbd5 protein in different mbd5 mutant alleles. (E, F) Locomotor behavioral analysis of the 7 dpf progeny from mbd5+/ΔMBD in-crosses upon treatment with 2.5mM Pentylenetetrazol (PTZ). The following numbers of individuals were obtained from the in-cross: mbd5+/+, N = 10; mbd5+/ΔMBD, N = 20; mbd5ΔMBD, N = 18. because some larvae did not move after PTZ treatment, the following numbers were used for data analysis: mbd5+/+, N = 7; mbd5+/ΔMBD, N = 20; mbd5ΔMBD, N = 17. (G) Locomotor defects in maternal-zygotic mbd5ΔMBD mutants and hyperactivity measured by average velocity after 2.5 mM PTZ treatment, N = 24. (H–K) Representative heat maps of adult ctrl (H, left) and mbd5ΔMBD (H, middle) zebrafish in a novel-tank assay virtually divided to top, middle and bottom zones (H, right). Locomotor behavioral analysis shows that the average distance moved (I) and average velocity (J) are significantly reduced in mbd5ΔMBD mutants comparing to controls. The time spent in different zones (K) shows no difference between mbd5ΔMBD mutants and controls. N = 13. Each value represents mean ± SD, ns, no significance, *P< 0.05, ** P< 0.01, *** P< 0.001.
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