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Nutrient restriction limits brain growth and NPC proliferation. (A) Diagram of the Xenopus optic tectum. NPCs reside along the ventricle at the midline (green), and their progeny are pushed laterally as NPCs divide and differentiate into neurons (red). The midline of the tectal ventricle is indicated with a dashed yellow line. (B) Experimental timeline for the data shown in C. Animals were treated for 24 h with CldU and then allowed to develop for 6 days in either fed or nutrient-restricted conditions. (C) Whole-mount confocal images of CldU immunofluorescence in the optic tectum in fed (i, iii) and nutrient-restricted (ii, iv) animals. 20× magnification 6.2 μm z-projections (i-ii) and 60× magnification 1.66 μm z-projections (iii-iv) are shown. Confocal images of brains were collected under identical settings and are shown at the same scale (i-ii and iii-iv). (D) Experimental timeline for the data shown in E. Animals were treated for 2 h with CldU and then allowed to develop for 2-3 days in either fed or nutrient-restricted conditions. (E) Whole-mount confocal images of Sox2 and CldU (i-ii), HuC/D and CldU (iii-iv), and HuC/D and pH3 (v-vi) immunofluorescence along the tectal midline in fed (i, iii, v) and nutrient-restricted (ii, iv, vi) animals. Single optical sections are shown. Scale bars: 100 μm (C); 20 μm (E). |
