Fig. 3: hcASD gene dyrk1a perturbation reduces ENCC migration and decreases gut motility in vivo.
From: Autism gene variants disrupt enteric neuron migration and cause gastrointestinal dysmotility
a dyrk1a perturbation through CRISPR injection, morpholino (MO) injection, or Dyrk1a inhibitor (TG003) treatment all reduce the area of ENCC migration in the gut compared to control CRISPR, control MO, or vehicle control (DMSO). b Area of gut migration quantification by condition. For injection comparisons, a two-tailed Wilcoxon matched-pairs signed rank test to compare the CRISPR or morpholino side to the control side within each animal was performed. Control CRISPR (N = 18, p = 0.7987), dyrk1a CRISPR (N = 18, p = 0.0032), Control MO (N = 24, p = 0.1208), dyrk1a MO (N = 41, p < 0.0001). For small molecule treatment, a two-tailed Wilcoxon rank sum test was performed to compare DMSO (N = 18) to 5 µM TG003-treated (N = 20) embryos, p = 0.0026. All samples are independent biological replicates from the same mating pair. In the box plot whiskers show minimum and maximum values, box represents the 25th and 75th quartiles, and the center line describes the median. c Schematic of the in vivo gut motility assay. At NF stage 47, tadpoles were fed food with fluorescent beads for 2 h, then placed in a 6-well plate in 3D-printed baskets with 20 animals/well for 3 h. Tadpoles are then removed by taking out baskets and leaving excrement with fluorescent beads behind, after which the plates were imaged to count the number of beads per well. Created in BioRender. McCluskey, K. (2025) https://BioRender.com/w82f112. d Developmental inhibition of Dyrk1a (TG003-treated beginning at NF 20) results in decreased fluorescent bead excretion compared to vehicle control (DMSO). Representative images of fluorescent beads (false-colored black). e Number of fluorescent beads per well counted for DMSO (N = 3 wells, 20 tadpoles each) and TG003 (N = 3 wells, 20 tadpoles each) (Dyrk1a inhibitor) wells. A one-tailed t test with Welch’s correction was used to compare DMSO to 5 µM TG003, p = 0.0091. Data are presented as mean values +/− SEM. Source data are provided as a Source Data file.
