Milan Jamrich

Cell interactions and the control of gene activity during early development of Xenopus laevis

During embryonic development, regulation of the zygotic genome may be mediated by inductive interactions and by cell-autonomous inheritance of informational material from the egg; we have studied certain aspects of such regulatory events in Xenopus laevis. Embryos cultured in Ca2+/Mg2+-free medium can be dissociated and dispersed, eliminating cell-cell contact and thus precluding inductive interactions. Such manipulations revealed that activation of the muscle-specific alpha-actin genes is absolutely dependent upon cell contacts. Conversely, the endoderm-specific DG42 gene and the ectoderm-specific DG81 cytokeratin gene are activated in embryo cells dispersed throughout blastula stages and therefore appear to be controlled by inherited factors. Nevertheless, abnormal cell arrangements may prevent expression of the cytokeratin gene, suggesting that animal pole cells can be diverted from their normal ectodermal fate by inductive contact with vegetally derived cells. The interactions required for alpha-actin induction and inhibition of cytokeratin expression are independent of strong adhesion between embryonic cells mediated by high concentrations of divalent cations.

Accumulation and decay of DG42 gene products follow a gradient pattern during Xenopus embryogenesis

The DG42 gene is expressed during a short window during embryogenesis of Xenopus laevis. The mRNA for this gene can be first detected just after midblastula, peaks at late gastrula, and decays by the end of neurulation. The sequence of the DG42 cDNA and genomic DNA predicts a 70,000-Da protein that is not related to any other known protein. Antibodies prepared against portions of the DG42 open reading frame that had been expressed in bacteria detected a 70,000-Da protein in the embryo with a temporal course of appearance and decay that follows that of the RNA by several hours. Localization of the mRNA in dissected embryos and immunohistochemical detection of the protein showed that DG42 expression moves as a wave or gradient through the embryo. The RNA is first detected in the animal region of the blastula, and by early gastrula is found everywhere except in the outer layer of the dorsal blastopore lip. By midgastrula DG42 protein is present in the inner ectodermal layer and the endoderm; it disappears from dorsal ectoderm as the neural plate is induced and later decays in a dorsoventral direction. The last remnants of DG42 protein are seen in ventral regions of the gut at the tailbud stage.

Transcription of a long, interspersed, highly repeated DNA element in Xenopus laevis

We have analyzed the transcription of 1723, a long, repeated DNA element that is interspersed in the genome of Xenopus laevis (B. K. Kay and I. B. Dawid (1983) J. Mol. Biol. 170, 583-596). We have detected RNA homologous to 1723 in total cellular RNA from ovaries, embryos, liver, and cultured kidney cells. Transcripts from both strands of the element are present at similar concentrations in these different RNA preparations. In oocytes, approximately 100 pairs of lampbrush chromosome loops are active in the transcription of 1723 elements. The abundance of 1723 RNA increases during embryogenesis, with the highest level reached at the tadpole stage. From cellular fractionation studies, we conclude that 1723 transcripts are largely limited to the nucleus.

Gene expression in amphibian embryogenesis

The study of molecular events during the embryogenesis of Xenopus laevis has advanced as a result of the availability of molecular markers, i.e., nucleic acid and antibody probes for genes that are expressed in a temporally and spatially regulated fashion during development. In this article we summarize results on the localized expression of keratin genes and on the reconstruction of regulated transcription of the gastrula/neurula-specific DG42 gene. Furthermore, we discuss experiments that investigate molecular events during mesoderm induction and provide information on the nature of the inducing principle.

Expression pattern of an axolotl floor plate-specific fork head gene reflects early developmental differences between frogs and salamanders.

Gastrulation is one of the most important stages of animal development and, as such, tends to be remarkably conserved. Therefore it is interesting to see that the two amphibian species, Xenopus laevis (frog) and Ambystoma mexicanum (axolotl), are different in the arrangement of cell types just before and during gastrulation. In Xenopus, the cells that will form dorsal mesoderm are located deep in the dorsal marginal zone, while in the axolotl, these are on the surface of the embryo. In this study we investigated whether homologous genes known to be involved in the formation of dorsal structures show a different pattern of expression in these two species. For this purpose, we isolated a fork head gene (AxFKH 1) from the axolotl, which is likely to be the homologue of the Xenopus fork head gene, XFKH 1 (Pintallavis, XFD-1). We find that AxFKH 1 and XFH 1 have a similar pattern of expression, but there are some important differences. In early gastrulae, transcripts are detected in the organizer region of both species. In late gastrulae, the transcripts in Xenopus are located in both the superficial and deep layers, but they are only found in the superficial layer of axolotl embryos. During neurulation, XFKH 1 is expressed in notochord and neural floor plate, whereas AxFKH 1 is expressed in the neural floor plate only. We propose that the differences in expression pattern of these two genes are due to a difference in formation of dorsal structures between these two species. Furthermore, the expression pattern of these two genes early in gastrulation is consistent with the idea that at least some of the neural floor plate cells are already determined at this time.

Chapter 24. The Role of Homeobox Genes in Vertebrate Embryonic Development

Publisher Summary This chapter discusses the role of homeobox genes in vertebrate embryonic development. The first vertebrate homeobox containing gene to be identified was isolated from xenopus. Subsequently, a large number of genes were isolated from three vertebrate species—xenopus, mouse, and humans. Approximately 40 mammalian genes have been isolated that contain homeobox sequences most similar to that of the Drosophila antp gene. These genes are collectively known as Hox genes and represent the largest group of vertebrate homeobox genes known to date. Many Hox gene family members are differentially expressed in the vertebrate limb bud, often with a graded pattern of expression. In xenopus, functional aspects of homeobox gene activity have been studied by ectopic expression or overexpression of these genes or by ablation of their gene products in embryos. Xenopus homeobox genes have been shown to be involved in pattern formation. Gain-of-function phenotypes have been similarly generated in mice, with ectopic expression and overexpression of homeobox gene constructs being achieved through the use of transgenic technology. Loss-of-function phenotypes can be obtained through the use of embryonic stem (ES) cells, in which genes can be mutated via homologous recombination. These gain of function and loss-of-function studies clearly illustrate the developmental importance of homeobox gene activity for proper vertebrate morphogenesis. There are, however, several naturally occurring mutations that have been identified in non- antp class homeobox genes. At least two known mouse mutations affecting morphogenesis have been shown to bear mutations in homeobox genes. There is also convincing evidence that aberrant homeobox gene expression can contribute to the genesis or progression of cancer. These data provide intriguing clues concerning the developmental role of the homeobox genes in vertebrates. It is likely that the further analysis of homeobox genes can implicate them in human disease and developmental malformation.