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Within the developing zebrafish retina, onset and progression of Neurolin\u2010a expression parallels the pattern of retinal ganglion cell (RGC) differentiation. By using a morpholino\u2010based knockdown approach, we show that Neurolin\u2010a (but not Neurolin\u2010b) is necessary for a crucial step in RGC differentiation. Without Neurolin\u2010a, a large proportion of RGCs fail to develop, and RGC axons are absent or reduced in number. Subsequently, Neurolin\u2010a is required for RGC survival and for the differentiation of all other retinal neurons. Neurolin\u2010b is expressed later in well\u2010differentiated RGCs and is required for RGC axon pathfinding. Without Neurolin\u2010b, RGC axons grow in highly aberrant routes along the optic tract and\/or fail to reach the optic tectum. Thus, the zebrafish Neurolin paralogs are involved in distinct steps of retinotectal development. J. Comp. 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