Complex Regulation of cyp26a1 Creates a Robust Retinoic Acid Gradient in the Zebrafish Embryo

Abstract

Positional identities along the anterior–posterior axis of the vertebrate nervous system are assigned during gastrulation by multiple posteriorizing signals, including retinoic acid (RA), fibroblast growth factors (Fgfs), and Wnts. Experimental evidence has suggested that RA, which is produced in paraxial mesoderm posterior to the hindbrain by aldehyde dehydrogenase 1a2 ( aldh1a2/ raldh2), forms a posterior-to-anterior gradient across the hindbrain field, and provides the positional information that specifies the locations and fates of rhombomeres. Recently, alternative models have been proposed in which RA plays only a permissive role, signaling wherever it is not degraded. Here we use a combination of experimental and modeling tools to address the role of RA in providing long-range positional cues in the zebrafish hindbrain. Using cell transplantation and implantation of RA-coated beads into RA-deficient zebrafish embryos, we demonstrate that RA can directly convey graded positional information over long distances. We also show that expression of Cyp26a1, the major RA-degrading enzyme during gastrulation, is under complex feedback and feedforward control by RA and Fgf signaling. The predicted consequence of such control is that RA gradients will be both robust to fluctuations in RA synthesis and adaptive to changes in embryo length during gastrulation. Such control also provides an explanation for the fact that loss of an endogenous RA gradient can be compensated for by RA that is provided in a spatially uniform manner.

Author Summary

The formation of gradients of morphogens, signaling molecules that determine cell fates in a concentration-dependent manner, is a fundamental process in developmental biology. Several morphogens pattern the anterior–posterior (head to tail) axis of the vertebrate nervous system, including the vitamin A derivative, retinoic acid (RA) and fibroblast growth factors (Fgfs). However, it remains unclear how the activities of such morphogen gradients are coordinated. We have addressed this question by combining genetic experiments in zebrafish and computational analyses. We show that RA acts as a graded signal over long distances and that its gradient is shaped, to a large extent, by local control of RA degradation. In particular, RA promotes and Fgf suppresses RA degradation, thereby linking the shapes of RA and Fgf gradients. Computational models suggest that this linkage helps make RA-mediated patterning robust to changes in the rate at which RA is synthesized (which may vary with levels of dietary vitamin A) as well as in the size and shape of the embryo during development. Analogous regulatory loops may be used for similar purposes in other tissues in which RA and Fgfs interact, as well as in other morphogen systems.

Abstract

Experimental and computational studies in zebrafish reveal a complex system regulating degradation of the vitamin A derivative retinoic acid along the anterior-posterior axis, which helps explain how morphogen gradients are established and maintained.