Anatomy of neurogenesis in the early zebrafish brain

The detailed architecture of postembryonic (i.e. secondary, as opposed to primary) neurogenesis in the zebrafish brain at 2 days postfertilization was investigated by studying expression domains of various proneural basic helix-loop-helix genes (i.e. neuroD=nrd, neurogenin1=ngn1, Zash-1b) and neurogenic genes (i.e. Notch-1a, deltaA) on the level of in situ-hybridized sectioned material and compared with brain sections of the same age immunostained for PCNA (a proliferation marker) or for Hu-proteins (marker for early neuronal differentiation). Whereas both Notch-1a and deltaA domains are present in all proliferative zones of the brain, only the more diffuse and scattered deltaA expression appears to extend additionally into the adjacent postmitotic gray matter. Zash-1b is the first achaete-scute orthologue shown here to be expressed exclusively in all proliferative central nervous zones (except for the eye). The ngn1 and nrd genes are typically both expressed in overlapping fashion in many-but not all-brain regions, with ngn1 being more restricted towards the ventricular proliferative zones and nrd extending more laterally. This fact as well as comparisons with PCNA- and Hu-immunostains indicate that a great proportion of ngn1-positive cells are mitotic, but some appear to extend into the postmitotic gray matter where the nrd-domains lie. A comparison of the relative extents of PCNA- (proliferative), nrd- (freshly determined) and Hu-positive (differentiating) cell populations allows to determine the relative maturation state of a given brain part. Similar to findings in late embryonic amniote brains, expression of nrd is absent (from 2 to 5 days) in the zebrafish subpallium, ventral preoptic region, ventral thalamus and hypothalamus. These four regions are also free of ngn1 expression (with the exception of an unusual peripheral ngn1 domain in the preoptic region), indicating that a neurogenetic network not involving nrd and ngn1 is at work there. Our characterization of locally distinct patterns and dynamics of secondary neurogenesis in the entire early (2 dpf) postembryonic zebrafish brain delivers the blueprint for more specialized functional studies.