Conservation of Pitx1 expression during amphibian limb morphogenesis.

In contrast to the pattern of limb emergence in mammals, chicks, and the newt N. viridescens, embryos such as Xenopus laevis and Eleutherodactylus coqui initiate pelvic limb buds before they develop pectoral ones. We studied the expression of Pitx1 in X. laevis and E. coqui to determine if this paired-like homeodomain transcription factor directs differentiation specifically of the hindlimb, or if it directs the second pair of limbs to form, namely the forelimbs. We also undertook to determine if embryonic expression patterns were recapitulated during the regeneration of an amputated limb bud. Pitx1 is expressed in hindlimbs in both X. laevis and E. coqui, and expression is similar in both developing and regenerating limb buds. Expression in hindlimbs is restricted to regions of proliferating mesenchyme.

CAP1 expression is developmentally regulated in Xenopus.

We have cloned and characterized a Xenopus member of the cyclase associated protein (CAP) gene family. xCAP1 is expressed as a maternal transcript, but is up-regulated prior to gastrulation and subsequently localizes to head mesenchyme, lens, otic vesicle, and trunk mesoderm including the pronephros. At different stages, the gene also appears to differentiate surface from deep (sensorial) ectoderm. As in Drosophila, Xenopus CAP1 is expressed in the developing eye, specifically in the differentiating lens. However, in distinction to Drosophila, Xenopus CAP1 does not express in periodically arrayed neural bands.

Xenopus adenine nucleotide translocase mRNA exhibits specific and dynamic patterns of expression during development.

We report the isolation and characterization of the Xenopus homolog to human T1 ANT (adenine nucleotide translocase). The 1290-nucleotide sequence contains initiation and termination signals, and encodes a conceptual protein of 298 amino acids. The sequence shares high amino acid identity with the mammalian adenine translocases. The transcript is present in unfertilized eggs, and it is expressed at higher levels during formation of the antero-posterior dorsal axis in embryos. Although low levels are expressed constitutively except in endodermal cells, adenine nucleotide translocase (ANT) expression is dynamically regulated during neurulation. At this stage, expression in ectoderm rapidly diminishes as the neural folds form, and then ANT expression increases slightly in mesoderm. At the culmination of neurulation, the neural tube briefly expresses ANT, and thereafter its expression predominates in the somitic mesoderm and also the chordoneural hinge. In addition, ANT expression is particularly high in the prosencephalon, the mesencephalon, the branchial arches, eye, and the otic vesicle. Treatment of embryos with retinoic acid has the effect of diminishing constitutive expression of ANT, but microinjection studies demonstrate that immediate and local repression cannot be induced in dorsal structures.

xPitx1 plays a role in specifying cement gland and head during early Xenopus development.

Xenopus Pitx1 is a homeobox gene whose family members are structurally and functionally conserved in organisms as diverse as Drosophila, chick, mouse, human, and frog. Present as a maternal transcript, the gene is zygotically expressed during gastrulation in a dorsal streak of cells. This streak restricts to a small circular domain underlying the center of presumptive neural plate. Shortly thereafter, a crescent of expression develops at the border of anterior neural ectoderm, and as the central plate domain diminishes, the crescent coalesces to define the presumptive cement gland. Expression remains high throughout cement gland development, and subsequently expands to include ectodermal cells involved in stomodeal invagination. During early organogenesis, expression ensues in developing eye, posterior lateral mesoderm, and first branchial arch derivatives. Ectopic expression of xPitx1 causes head deformities including enlarged cement gland, ectopic cement glands, and posterior deformities or, in extreme cases, inhibition of recognizable structures posterior to the cement gland. Expression of markers such as XCG-1, xOtx2, xPax6, neuralbeta tubulin, and xTwist suggest that increases in cement gland and lower mandibular size are likely at the expense of other head tissues. Paradoxically, overexpression is sufficient to partially rescue embryos that are axially perturbed by ultraviolet irradiation or retinoic acid administration. Ectopic expression of xPitx1 in ectodermal explants directly promotes cement gland development as there was no evidence that mesodermal or neural tissue was present in explants.