Extended Data Fig. 3: Structural changes in the piglet Arc during perinatal stages.
a, Schematic comparing the stages of human and pig development15,16. PCD, post-conception day. b, Nissl stain of coronal section from piglet brains across ages. Scale bars, 3 mm. c, Schematic of the P0 piglet brain. The LV of the left hemisphere is highlighted in gray (left) and the Arc in green. Lateral ventricle (LV). d, The 3D volume of the Arc was measured from Nissl-stained piglet serial sections. e, Top: schematic showing piglet Arc subregions at coronal planes. Down: quantification of Arc area at each plane across ages. Two-tailed unpaired t test and data are presented as the mean ± s.e.m. (n = 3 cases). Olfactory ventricle (OV); anterior Arc (A-Arc); posterior Arc (P-Arc). Sample size is provided as source data. f, Confocal images of core Arc structural changes across ages. The yellow line highlights the boundary between tiers 1 and 2; the white line between tiers 3 and 4. Scale bars, 500 µm. This experiment has been repeated five times. g, Top: confocal image of DCX+ expression in developing piglet Arc. Arrows indicate blood vessels. Scale bars, 500 µm. Bottom: quantification of DCX expression pixel intensity across tiers. Data are presented as the mean ± s.e.m. (n = 3 cases). Sample size is provided as source data. h, Quantification of tier thickness in developing piglet brains. Scale bar, 500 µm. Two-tailed unpaired t tests and data as the means ± s.e.m. i–l, Confocal images of DCX+ cells near α-SMA+ blood vessels (BV) in tier 3 of the piglet Arc across ages. Scale bar, 30 µm. Quantification of the BV size (j), the total number of DCX+ cells in contact with the BV (k) and the density of DCX+ cells contacting with (w) and without (w/o) BV across ages (l). Two-tailed unpaired t tests and data as mean ± s.e.m. (n = 3 cases). Sample size and P values are provided as source data. m, Schematic showing Arc features age-matched between humans and piglets (blue lines) and the period when the tiered Arc structure is present in human and piglet brains (yellow lines).
