Ken W.Y. Cho

Molecular nature of Spemann's organizer: the role of the Xenopus homeobox gene goosecoid

This study analyzes the function of the homeobox gene goosecoid in Xenopus development. First, we find that goosecoid mRNA distribution closely mimics the expected localization of organizer tissue in normal embryos as well as in those treated with LiCl and UV light. Second, goosecoid mRNA accumulation is induced by activin, even in the absence of protein synthesis. It is not affected by bFGF and is repressed by retinoic acid. Lastly, microinjection of goosecoid mRNA into the ventral side of Xenopus embryos, where goosecoid is normally absent, leads to the formation of an additional complete body axis, including head structures and abundant notochordal tissue. The results suggest that the goosecoid homeodomain protein plays a central role in executing Spemanns organizer phenomenon.

The homeobox gene goosecoid controls cell migration in Xenopus embryos

Goosecoid (gsc), a homeobox gene expressed specifically in the dorsal blastopore lip of the Xenopus gastrula, is considered to play an important role in Spemanns organizer phenomenon. Lineage tracing and time-lapse microscopy were used to follow the fate of embryonic cells microinjected with gsc mRNA. Microinjected gsc has non-cell autonomous effects, recruiting neighboring uninjected cells into a twinned dorsal axis. Ectopic expression of gsc mRNA in ventral blastomeres as well as overexpression of gsc in dorsal blastomeres leads to cell movement toward the anterior of the embryo. The results suggest a function for gsc in the control of gastrulation movements in groups of cells, but not in dissociated cells, and demonstrate that a vertebrate homeobox gene can regulate region-specific cell migration.

Differential utilization of the same reading frame in a Xenopus homeobox gene encodes two related proteins sharing the same DNA-binding specificity.

Xenopus XlHbox 1 produces two transcripts during early development. One encodes a long open reading frame (ORF) and the other a short ORF sharing the same homeodomain, but differing by an 82 amino acid domain at the amino terminus. The long protein amino terminus is conserved with many other homeodomain proteins, and its absence from the short protein could have functional consequences. Some viral genes also utilize a single ORF to encode transcription factors of antagonistic functions. The overall organization of the homologous genes in frog and man is similar, supporting the notion that both transcripts are of functional significance. Studies on XlHbox 1 function show that the region common to the long and short proteins has a sequence‐specific DNA‐binding activity, and that microinjection of specific antibodies into embryos results in the loss of structures derived from cells normally expressing XlHbox 1.

Overexpression of a homeodomain protein confers axis-forming activity to uncommitted xenopus embryonic cells

The anteroposterior character of mesoderm induced by a peptide growth factor (XTC-MIF) was tested by transplantation into host Xenopus gastrulae. Both retinoic acid and a homeodomain protein were able to override the anteriorizing effect of the growth factor. Microinjection of a posteriorly expressed homeobox mRNA can respecify anteroposterior identity, transforming head mesoderm into tail-inducing mesoderm. Unexpectedly, overexpression of XIHbox 6 protein in the transplanted cells, without addition of growth factors, caused the formation of tail-like structures. The cells overexpressing XIHbox 6 were able to recruit cells from the host into the secondary axis. The results suggest that vertebrate homeodomain proteins are part of the biochemical pathway leading to the generation of the body axis.

Duplicated homeobox genes in Xenopus

Multiple kinds of clones and restriction fragment polymorphisms are frequently encountered when analyzing genes of the tetraploid frog Xenopus laevis. Two types of cDNA clone have been isolated for homeobox gene 2. Analysis of their corresponding genomic clones confirmed the existence of clearly distinct restriction maps; in addition the nearby presence of two additional homeoboxes suggests that this region is homologous to the Hox 2 gene complex of mammals. We asked whether the genetic polymorphism in Xenopus results from increased allelic differences due to tetraploidy or from having duplicated Hox 2 complexes. Using X. laevis/Xenopus borealis interspecies hybrids we show that the two types of X. laevis homeobox gene 2 transcripts result from two different genetic loci. They cannot represent alleles of the same gene because they do not segregate independently in the F1 hybrid progeny. Most other X. laevis homeobox genes studied so far are also found in two versions. Thus X. laevis seems to have two homeobox genes, both of which are expressed, for each one present in mammals or other vertebrates.