Fig. 2

Fig. 2 The rescuing effect of methocarbamol relies on its primary pharmacological target, the CA family.

a, Proportion of morphological abnormalities in fish with pan-neuronal expression of Dendra–tau-A152T after treatment with 1 μM methazolamide (MTZ), 0.3 μM acetazolamide (Aceta.) or 1 μM tioxolone (Tiox.). Treatment with all CA inhibitors resulted in phenotypic rescue compared with DMSO-treated siblings (n = 6 clutches with ≥20 fish in each group; ***P ≤ 0.001 versus DMSO. Data were analyzed by χ² test. b,c, Representative images of the retina of transgenic rho:eGFP–tau-WT and rho:eGFP–tau-P301L fish (b) and quantification of the pixel area of eGFP–tau⁺ photoreceptors following treatment with 1 μM methazolamide (c), showing an increase in eGFP–tau⁺ photoreceptors after treatment with methazolamide relative to DMSO-treated siblings (n ≥ 32 eyes per condition; the graph shows mean ± s.d.). Data were analyzed by one-way ANOVA, ^####P≤ 0.0001, followed by Tukey’s multiple comparisons test with the following significance values: ***P ≤ 0.001 versus DMSO (–) for wild-type tau and **P ≤ 0.01 versus DMSO (–) for tau-P301L. Scale bar, 50 μm. d, Quantification of the pixel area of eGFP–tau⁺ rod photoreceptors following CRISPR injection. CRISPR targeting of cahz, ca4a, ca9 and ca14 increased the area of rod photoreceptors relative to uninjected siblings (for representative images, see Extended Data Fig. 4e; n ≥ 39 eyes per condition; data are shown as mean ± s.d.); *P ≤ 0.05, **P ≤ 0.01 and ***P ≤ 0.001 versus uninjected. Data were analyzed by two-tailed unpaired t-test. e, Proportion of morphological abnormalities in fish with pan-neuronal expression of Dendra–tau-A152T following CRISPR-based knockdown of CA isoforms. Genetic inhibition of ca4a, ca5, ca9 and ca14 increased the proportion of normal phenotypes, whereas CRISPR injection targeting cahz and ca2 worsened the morphological defects compared with uninjected siblings (n = 3 clutches for toxic conditions (cahz and ca2) and n ≥ 4 clutches with a minimum of 30 fish per clutch for ca4a, ca5, ca9 and ca14); ****P ≤ 0.0001 versus uninjected. Data were analyzed by χ² test.

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