César Arenas-Mena

The A/P axis in echinoderm ontogeny and evolution: evidence from fossils and molecules

SUMMARY Even though echinoderms are members of the Bilateria, the location of their anterior/posterior axis has remained enigmatic. Here we propose a novel solution to the problem employing three lines of evidence: the expression of a posterior class Hox gene in the coeloms of the nascent adult body plan within the larva; the anatomy of certain early fossil echinoderms; and finally the relation between endoskeletal plate morphology and the associated coelomic tissues. All three lines of evidence converge on the same answer, namely that the location of the adult mouth is anterior, and the anterior/posterior axis runs from the mouth through the adult coelomic compartments. This axis then orients the animal such that there is but a single plane of symmetry dividing the animal into left and right halves. We tentatively hypothesize that this plane of symmetry is positioned along the dorsal/ventral axis. These axis identifications lead to the conclusion that the five ambulacra are not primary body axes, but instead are outgrowths from the central anterior/posterior axis. These identifications also shed insight into several other evolutionary mysteries of various echinoderm clades such as the independent evolution of bilateral symmetry in irregular echinoids, but do not elucidate the underlying mechanisms of the adult coelomic architecture.

Hindgut specification and cell-adhesion functions of Sphox11/13b in the endoderm of the sea urchin embryo

Sphox11/13b is one of the two hox genes of Strongylocentrotus purpuratus expressed in the embryo. Its dynamic pattern of expression begins during gastrulation, when the transcripts are transiently located in a ring of cells at the edge of the blastopore. After gastrulation, expression is restricted to the anus–hindgut region at the boundary between the ectoderm and the endoderm. The phenotype that results when translation of Sphox11/13b mRNA is knocked down by treatment with morpholino antisense oligonucleotides (MASO) suggests that this gene may be indirectly involved in cell adhesion functions as well as in the proper differentiation of the midgut–hindgut and midgut–foregut sphincters. The MASO experiments also reveal that Sphox11/13b negatively regulates several downstream endomesoderm genes. For some of these genes, Sphox11/13b function is required to restrict expression to the midgut by preventing ectopic expression in the hindgut. The evolutionary conservation of these functions indicates the general roles of posterior Hox genes in regulating cell‐adhesion, as well as in spatial control of gene regulatory network subcircuits in the regionalizing gut.

Embryonic expression of HeFoxA1 and HeFoxA2 in an indirectly developing polychaete

Two forkhead family transcription factors, HeFoxA1 and HeFoxA2, were isolated from the serpulid annelid Hydroides elegans and their transcript distribution were characterized during embryogenesis. HeFoxA1 is first detected in second quartet blastomeres soon after their formation, and later in all vegetal half blastomeres, which comprise ectoderm, endoderm, and mesoderm precursors. HeFoxA1 expression declines first in subtrochal ectoderm and presumptive midgut precursors, as well as apparently in D quadrant blastomeres in advance of any known signaling events. Later, during gastrulation, HeFoxA1 declines in hindgut precursors, and by the end of gastrulation the expression remains active only in foregut precursors. HeFoxA1 is apparently expressed in ectomesoderm cells involved in forming the larva-specific protonephridium (the so-called head kidney). The other ortholog, HeFoxA2, is expressed in a subset of the cells in which HeFoxA1 is expressed during early stages, but later it is largely restricted to the endoderm–ectoderm boundary of the proctodaeum. In addition, HeFoxA2 has a unique expression in two hindgut cells and abutting ectoderm cells located by the imminent anal opening. The combined expression of HeFoxA1 and HeFoxA2 correlates with mesoderm and endoderm expression of their orthologs in other bilaterians.

HeOtx expression in an indirectly developing polychaete correlates with gastrulation by invagination

The expression of an Otx homolog in the indirectly developing polychaete Hydroides elegans was characterized during embryo, trochophore, and feeding-larva stages. In the animal hemisphere, HeOtx is first expressed in 1q12 blastomeres and their immediate descendants. Such discrete embryonic animal hemisphere Otx expression perhaps relates to cell-type specification functions of the larva. During feeding stages, transcripts are detected in adult cerebral ganglia precursors and putative adult eye precursors, where it may have adult brain regionalization functions. HeOtx is not expressed in primary trochoblast precursors, but it is expressed in cells adjacent to the ciliary band. HeOtx is also expressed in a group of cells in the dorsal midline of the early trochophore larva in putative posterior sensory organ precursors. The vegetal hemisphere expression starts in oral and lateral sides of the blastopore and later expands to central blastomeres that lead the gastrulation movements. During late gastrulation stages, the expression declines in foregut precursors, but it is maintained in midgut precursors, suggesting its involvement in tripartite gut subdivision functions. HeOtx broader and earlier endoderm expression correlates with gastrulation by invagination associated with the formation of the feeding trochophore, in contrast with a later and orally restricted Otx expression found in a polychaete that gastrulates by epiboly and forms a non-feeding trochophore. The endoderm expression and functional roles in other bilaterians suggest an ancestral role of Otx related to gastrulation by invagination.

Sinistral equal‐size spiral cleavage of the indirectly developing polychaete Hydroides elegans

Two major variants of the stereotypic spiral cleavage correlate with distinct developmental modes in polychaetes. Indirect development through a feeding trochophore larva correlates with development from four equal‐sized blastomeres, whereas direct development correlates with unequal cleavage characterized by a large dorsal blastomere precursor maternally predetermined. The equal‐size spiral cleavage of the indirectly developing serpulid Hydroides elegans has been reconstructed from serial sections of nuclei‐stained embryos. The order of cell divisions has been determined from the 2‐cell stage to the 80‐cell stage, when gastrulation cell movements start to overlap with late spiral‐cleavage divisions. In contrast to related species, the third cleavage in Hydroides elegans is invariably sinistral. The four quadrants remain indistinct until the 60‐cell stage, when the small 2d22 and large 2d21 cells are generated. The developmental significance of the invariant spiral cleavage relates to the spatial distribution of gene functions that it partitions and their relation to blastomere fate commitments. The conservation and divergence of the cleavage pattern among spiralians is well suited to study the developmental control of the cell‐cleavage machinery and its evolution. Developmental Dynamics 236:1611–1622, 2007.

Histone H2A.Z expression in two indirectly developing marine invertebrates correlates with undifferentiated and multipotent cells

SUMMARY The embryos of indirect developers generate an intermediate larval stage that nourishes the proliferation of undifferentiated multipotent cell precursors in charge of postembryonic adult formation. Multipotency affects the regulation of many genes and seems to be mediated in part by chromatin modification. Chromatin transcriptional properties are regulated by histone modification and by incorporation of peculiar histone variants. The histone variant H2A.Z is associated with transcriptionally competent chromatin and silent genes primed for activation or permanent repression. However, despite the extensive mechanistic characterizations in unicellular eukaryots, the essential role of the highly conserved H2A.Z variant during animal embryogenesis remains obscure. We show that the expression of H2A.Z in the larvae of two distant indirectly developing marine invertebrates, a polychaete and a sea urchin, remains high in all their embryonic and postembryonic developmentally competent cell precursors, and declines during their differentiation. In particular, the expression in undifferentiated multipotent adult precursors during feeding larval stages in both organisms provides unique insight about its general association with developmental potential. Our experiments confirm previous reports indicating that the expression of H2A.Z is proliferation (DNA synthesis) independent, in contrast with the DNA synthesis dependence of “mainstream” histones. We suggest that similar H2A.Z transcriptional functions previously identified in unicellular organisms also help to maintain an open chromatin state competent for transcriptional‐regulatory transactions during metazoan development.

The transcription factors HeBlimp and HeT-brain of an indirectly developing polychaete suggest ancestral endodermal, gastrulation, and sensory cell-type specification roles.

The expression of Blimp and T-brain was characterized during embryogenesis of the indirectly developing polychaete Hydroides elegans. The expression of both genes in the vegetal blastomeres of this lophotrochozoan is restricted to the lateral and oral regions of the blastopore. Both transcription factors also have expression patterns consistent with ancestral neural functions. HeBlimp is expressed in a couple of animal cap cells and exhibits a marked left bias that correlates with earlier development of the left eye in H. elegans. HeT-brain is expressed in animal cap blastomeres and eventually becomes restricted to apical tuft cells of early trochophore stages. The expression of Blimp and T-brain among directly and indirectly developing bilaterians suggests ancestral sensory cell-type specification, gastrulation by invagination, endoderm specification, mesoderm specification, and/or tripartite gut subdivision. These characterizations add to the long-term goal of understanding the regulatory evolution functions that underlies complex life cycles in metazoans.

Developmental transcriptional‐competence model for a histone variant and a unicellular origin scenario for transcriptional‐multipotency mechanisms

The advent of multicellularity had more to do with evolutionary transformations favoring differential gene expression among constituent cells than with keeping cells together after mitosis. Differential gene expression is mainly controlled at the transcriptional level because transcriptional regulatory DNA has enormous parallel processing power; that is, the transcriptional regulatory DNA of each gene can simultaneously process multiple temporal and/or spatial upstream regulatory inputs from the transcription factors for which it has binding sites. Cell differentiation in metazoans is accompanied by a decline in differential gene expression potential. Thus, in contrast with unicellular organisms, the differentiated cell of a multicellular organism can only express a subset of the genes in the genome. The stability of the transcriptional states associated with terminal differentiation could have evolved by losing the ability to reversibly shift among various transcriptional states associated with temporal adaptations in unicellular organisms. In complex metazoans, the spatial deployment of the gene-regulatory networks that control alternative transcriptional states associated with differentiation does not unfold at once but after several transitional phases during development. In principle, the same mechanisms allowing the unicellular transcriptional flexibility associated with the shift among temporal adaptations could be transiently used to allow the unfolding of diverse regulatory states during metazoan development. At the end of development, loosing the flexibility to shift among transient developmental transcriptional states would result in higher stability of the differentiation transcriptional states. The simultaneous occurrence of undifferentiation and differentiation processes in discrete embryonic domains of indirect developers is well suited to provide unique insight about the contrasted molecular mechanisms for multipotency transcriptional flexibility and differentiation transcriptional stability. A MODEL FOR THE PECULIAR TRANSCRIPTIONAL FUNCTIONS AND DEVELOPMENTAL EXPRESSION OF A HISTONE VARIANT