Fig. 5
OXR1 deficiency severely impairs brain development in distinct human brain organoid models. a The progression of cerebral organoid development is illustrated (upper). Representative bright-field images showing the progression of cerebral organoids derived from healthy control (CTRL) and patient (OXR1ΔEx18) iPSC clones (lower left). Black arrows point the expanding neuroepithelial formation in CTRL clones from day 8 and the poorly developed neuroepithelia buds observed in the OXR1ΔEx18 cerebral organoids at day 15. The body size of cerebral organoids (lower right) is quantified by summing the products of bi- dimensional measurements (SPD). SPD = Longest diameter (LD) × longest diameter perpendicular to the LD. n = 15–20 organoids of each clone, * p < 0.05. b Immunohistochemical analyses of cerebral organoids at day 75 using the antibody specific for MAP2 (the marker of mature neurons, yellow) or TBR1(the marker of deep layer neurons, magenta). c Immunohistochemical analyses of cerebral organoids at day 75 using antibodies specific for REELIN (yellow) and OXR1 (magenta). d A schematic illustration showing the guided progression of region-specific brain organoids. e Representative bright-field images showing the progression of hypothalamus derived from CTRL and OXR1ΔEx18 iPSCs. The starting cell number of each clone is indicated (100% refer to 10,000 iPSC cells per microwell of Aggrewell-800 plate on day 0). After hypothalamus patterning, the spheroids of OXR1ΔEx18 clone 1 (c1) and OXR1ΔEx18 clone 2 (c2) dissipated by day 5. f Representative bright-field images showing the progression of hippocampus organoids. After hippocampus patterning, the spheroids of OXR1ΔEx18 c2 dissipated by day 5. g Representative confocal images showing the progression of SOX2+ neural progenitor cells (magenta) during the development of hippocampus organoids. h Representative confocal images showing the progression of NeuN+ mature neurons (grey) and DCX+ immature neurons (green) during the development of hippocampus organoids. In all confocal images, the scale bar is 60 μm and the nuclei were visualized using DAPI (blue)