Loss of tmem184ba results in neuroanatomical defects, and in vivo complementation shows that some disease-associated variants result in loss of function (A) Representative bright-field lateral images of 3-dpf larvae injected with MO alone or co-injected with human TMEM184B mRNA (WT, p.Val88Met, p.Leu127Arg, p.Gly162Arg, p.Lys184Glu, p.Gly288Ala, and p.Arg282His). p.Arg282His is a presumed benign negative control (dbSNP: rs370107940; 215 heterozygotes and four homozygotes are present in 1,609,748 individuals in gnomAD v.4.1.0). Scale bar, 300 μm. (B and C) Quantification of the (B) lateral head size and (C) body length in 3-dpf larvae as indicated with anatomical landmarks in Figure 3A. Experimental conditions were normalized and presented as a percentage of the control mean. n = 50–88/condition. (D) Representative dorsal images of 3-dpf larval heads immunostained with acetylated tubulin. White dotted shape, optic tecta; white dotted line, midline between optic tecta where intertectal commissural axons were counted. Zoomed image at right of each panel corresponds to white box around both optic tecta. Anterior, left; posterior, right; Scale bar, 50 μm. (E) Quantification of optic tecta size as shown in (D). (F) Quantification of intertectal neurons that cross the dorsal midline as shown in (D). For (E) and (F), n = 30–47/condition. For (B), (C), (E), and (F), statistical analyses were performed with a one-way ANOVA with Tukey’s post-hoc test: ∗∗∗∗p < 0.0001; ∗∗∗p < 0.001; ∗∗p < 0.01; ns, not significant. Error bars represent standard deviation of the mean. Data point color: black, uninjected control; blue, WT mRNA; red, variant mRNA identified in affected individuals; green, presumed benign variant mRNA from gnomAD.
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