nestin is essential for zebrafish brain and eye development through control of progenitor cell apoptosis巢蛋白对斑马鱼通过控制大脑和眼睛发育至关重要的祖细胞凋亡.pdfVIP

Nestin Is Essential for Zebrafish Brain and Eye Development through Control of Progenitor Cell Apoptosis 1 2 1 Hua-Ling Chen , Chiou-Hwa Yuh , Kenneth K. Wu * 1 Institute of Cellular and System Medicine, National Health Research Institutes, Zhunan, Taiwan, 2 Division of Molecular and Genomic Medicine, National Health Research Institutes, Zhunan, Taiwan Abstract Background: Nestin is expressed in neural progenitor cells (NPC) of developing brain. Despite its wide use as an NPC marker, the function of nestin in embryo development is unclear. Methodology/Principal Findings: As nestin is conserved in zebrafish and its predicted sequence is clustered with the mammalian nestin orthologue, we used zebrafish as a model to investigate its role in embryogenesis. Injection of nestin morpholino (MO) into fertilized eggs induced time- and dose-dependent brain and eye developmental defects. Nestin morphants exhibited characteristic morphological changes including small head, small eyes and hydrocephalus. Histological examinations show reduced hind- and mid-brain size, dilated ventricle, poorly organized retina and underdeveloped lens. Injection of control nestin MO did not induce brain or eye changes. Nestin MO injection reduced expression of ascl1b (achaete-scute complex-like 1b), a marker of NPCs, without affecting its distribution. Nestin MO did not influence Elavl3/4 (Embryonic lethal, abnormal vision, Drosophila-like 3/4) (a neuronal marker), or otx2 (a midbrain neuronal marker), but severely perturbed cranial motor nerve development and axon distribution. To determine whether the developmental defects are due to excessive NPC apoptosis and/or reduced NPC proliferation, we analyzed apoptosis by TUNEL assay and acridi