Journal: Nature Communications
Article Title: Ependymoglial cells are critical for cortex regeneration in axolotls
doi: 10.1038/s41467-026-68538-6
Figure Lengend Snippet: a The timeline of BrdU/EdU administration post-MTZ-induced ablation in NeuroD6:Cherry-NTR2.0 (15 cm body length), with analysis at 113 days post-treatment (dpt). b Immunofluorescence images showing the distribution of BrdU + (green), EdU + (magenta), and SOX2 + (white) cells combined with DAPI (blue) in the regenerating dorsal pallium. c Quantification of labeled cell proportions in Z1 and Z2 of the regenerating dorsal pallium. Five or six serial sections from equivalent positions of telencephalic regions were analyzed ( n = 3). Each dot represents the percentage of each indicated cell population among total cells for an individual animal. d The timeline of BrdU/EdU administration in d/d (2–2.3 cm body length) during development, with analysis at 122 day after the first injection. e Immunofluorescence images showing the distribution of BrdU + (green), EdU + (magenta), and SOX2 + (white) cells combined with DAPI (blue) in the developing dorsal pallium. f Quantification of labeled cell proportions in Z1 and Z2 of the developing dorsal pallium. Five serial sections from equivalent positions of telencephalic regions were analyzed ( n = 3). Each dot represents an individual animal. g–x Immunofluorescence analysis of regenerated neuron subtype in NeuroD6:Cherry-NTR2.0 at 113 dpt. Co-labeling of BrdU + CHERRY + with pan-neuronal marker NeuN ( g – l ), restricted area markers CALB2 ( m–r ) and SATB1 ( s–x ) indicates regeneration of subtype-specific neurons in NeuroD6:Cherry-NTR2.0 , as shown in magnified single-channel and merged images. The yellow arrowheads indicate the triple-positive cells. y ISH analysis of pallial neuron markers ( Tmie , Tbr1 , Nptx1 , Fezf2 ) in NeuroD6:Cherry-NTR2.0 at 113 dpt. z Quantification of the dorsal pallium thickness in the MTZ and DMSO groups at 113 dpt. Five serial sections from equivalent positions of telencephalic regions were analyzed in MTZ or DMSO treated animals ( n = 5 each). Data were analyzed by unpair two-tailed Student’s t -test and represented as mean ± SEM. **** p < 0.0001. Source data are provided as a file. The red double-headed arrow indicates the measurement location for dorsal pallium thickness. White matter (WM), Zones Z1 (outer), Z2 (inner) and ventricular zone (VZ) are manually delineated. The yellow or black dashed lines outline the lumen surface of the brain. The white or black dashed lines outline the shape of brain or boundary of Z1/Z2 zone. Abbreviations: LV lateral ventricle. Scale bars: 100 μm in ( b ) and e; 200 μm in g , m , s , y .
Article Snippet: The following primary antibodies were used against the corresponding antigens: SOX2 (1:500, Abcam, ab97959; 1:200, Abcam, 14-9811-82), NeuN (1:300, Sigma, MAB377), TUJ1 (1:500, R&D, MAB1195), Cleaved Caspase-3 (1:500, CST, 9661 s), BrdU (1:500, Invitrogen, MA3-071; 1:500, Abcam, ab1893), CTIP2 (1:500, Abcam, ab18465), CALB2 (1:500, Swant, 7697), SATB1 (1:20, Abcam, ab51502), Iba1 (1:500, Wako, 019-19741), CHERRY (1:1000, Invitrogen, M11217 ), NKX2.2 (1:100, BD Biosciences,564731).
Techniques: Immunofluorescence, Labeling, Injection, Marker, Two Tailed Test
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