Pou-2--a zebrafish gene active during cleavage stages and in the early hindbrain

We have cloned the zebrafish pou-2 gene which encodes a novel type (class VII) of POU domain. Maternal pou-2 transcripts are initially found in all blastomeres. However, during later cleavage stages pou-2 expression disappears in the marginal cells. Some of their progeny will form the first lineage restricted compartment during zebrafish development. Blastula pou-2 expression in confined exclusively to the deep embryonic layer (DEL) forming the embryo proper. No expression is found in extraembryonic tissues, i.e. the yolk syncytial layer (YSL) and the enveloping layer (EVL). Thus pou-2 expression during early embryogenesis correlates with the continuing absence of cell lineage restriction. Towards the end of gastrulation, pou-2 expression becomes confined to the neural plate, predominantly to the prospective hindbrain and to the spinal cord. pou-2 expression in the forming hindbrain is restricted to future rhombomeres r2 and r4. Retinoic acid treatment during epiboly alters the hindbrain domains of pou-2, suggesting that the entire anterior hindbrain acquires r4-like properties. This finding is supported by analysis of early pax-2 and krx-20 expression patterns in RA-treated zebrafish embryos. The changes resemble similar hindbrain transformations observed in other vertebrates, supporting an evolutionary conservation of the mechanisms segmenting the hindbrain of vertebrates. pou-2 appears to respond to the same signals as other presumed patterning genes. This observation, together with pou-2 expression in the hindbrain prior to morphological segmentation, suggests an important role for this putative transcription factor in establishing and specifying rhombomeric segments.