RESUMO
Blindness afflicts ~39 million people worldwide. Retinal ganglion cells are unable to regenerate, making this condition irreversible in many cases. Whole-eye transplantation (WET) provides the opportunity to replace diseased retinal ganglion cells, as well as the entire optical system and surrounding facial tissue, if necessary. Recent success in face transplantation demonstrates that this may be a promising treatment for what has been to this time an incurable condition. An animal model for WET must be established to further enhance our knowledge of nerve regeneration, immunosuppression, and technical aspects of surgery. A systematic review of the literature was performed to evaluate studies describing animal models for WET. Only articles in which the eye was completely enucleated and reimplanted were included. Study methods and results were compared. In the majority of published literature, WET can result in recovery of vision in cold-blooded vertebrates. There are a few instances in which mammalian WET models demonstrate survival of the transplanted tissue following neurovascular anastomosis and the ability to maintain brief electroretinogram activity in the new host. In this study we review in cold-blooded vertebrates and mammalian animal models for WET and discuss prospects for future research for translation to human eye transplantation.
Assuntos
Cegueira/reabilitação , Olho/transplante , Traumatismos do Nervo Óptico/complicações , Retina/fisiologia , Animais , Modelos Animais de Doenças , Olho/fisiopatologia , Traumatismos do Nervo Óptico/fisiopatologia , Transplante de Órgãos/métodos , Transplante de Órgãos/tendências , Sobrevivência de Tecidos/fisiologiaRESUMO
The eumetazoan clade of modern animals includes cnidarians, acoels, deuterostomes, and protostomes. Stem group eumetazoans evolved in the late Neoproterozoic, possibly before the Marinoan glaciation, according to a variety of different kinds of evidence. Here, we combine this evidence, including paleontological observations, results from molecular and morphological phylogeny, and paleoecological considerations, with deductions from the organization of the gene regulatory networks that underlie development of the bilaterian body plan. Eumetazoan body parts are morphologically complex in detail, and modern knowledge of gene regulatory network structure shows that the control circuitry required for their development is hierarchical and multilayered. Among the consequences is that the kernels of the networks that control the early allocation of spatial developmental fate canalize the possibilities of downstream evolutionary change, a mechanism that can account for the appearance of distinct clades in early animal evolution. We reconstruct preeumetazoan network organization and consider the process by which the eumetazoan regulatory apparatus might have been assembled. A strong conclusion is that the evolutionary process generating the genomic programs responsible for developmental formulation of basic eumetazoan body plans was in many ways very different from the evolutionary changes that can be observed at the species level in modern animals.
Assuntos
Evolução Biológica , Morfogênese/genética , Animais , Padronização Corporal/genética , DNA/genética , Evolução Molecular , Fósseis , Regulação da Expressão Gênica no Desenvolvimento , Redes Reguladoras de Genes , Modelos Genéticos , FilogeniaRESUMO
The primary mesenchyme cells (PMCs) of the sea urchin embryo have been an important model system for the analysis of cell behavior during gastrulation. To gain an improved understanding of the molecular basis of PMC behavior, a set of 8293 expressed sequenced tags (ESTs) was derived from an enriched population of mid-gastrula stage PMCs. These ESTs represented approximately 1200 distinct proteins, or about 15% of the mRNAs expressed by the gastrula stage embryo. 655 proteins were similar (P<10(-7) by BLAST comparisons) to other proteins in GenBank, for which some information is available concerning expression and/or function. Another 116 were similar to ESTs identified in other organisms, but not further characterized. We conservatively estimate that sequences encoding at least 435 additional proteins were included in the pool of ESTs that did not yield matches by BLAST analysis. The collection of newly identified proteins includes many candidate regulators of primary mesenchyme morphogenesis, including PMC-specific extracellular matrix proteins, cell surface proteins, spicule matrix proteins and transcription factors. This work provides a basis for linking specific molecular changes to specific cell behaviors during gastrulation. Our analysis has also led to the cloning of several key components of signaling pathways that play crucial roles in early sea urchin development.
Assuntos
Expressão Gênica , RNA Mensageiro , Ouriços-do-Mar/genética , Animais , Clonagem Molecular , Etiquetas de Sequências Expressas , Proteínas da Matriz Extracelular/genética , Hibridização In Situ/métodos , Mesoderma , Ouriços-do-Mar/embriologia , Análise de Sequência de DNARESUMO
The CyIIa gene of the sea urchin embryo is a model for study of cis-regulation downstream of cell-type specification, as CyIIa transcription follows the specification and initial differentiation of the embryonic domains in which it is expressed. These are the skeletogenic and secondary mesenchyme and gut. We carried out a detailed structural and functional analysis of a cis-regulatory region of this gene, extending 780 bp upstream and 125 bp downstream of the transcription start site, that had been shown earlier to reproduce faithfully the complex and dynamic CyIIa pattern of expression. This analysis revealed that the overall pattern of expression of the CyIIa gene appears to be governed mainly by two independent sets of DNA elements, which are target sites for specific proteins present in blastula-stage nuclear extract. One type of element, which controls a dynamic program of expression in both skeletogenic and secondary mesenchyme cells, contains the consensus-binding site for a member of the ets transcription factor family. The other, which is responsible for the terminal or permanent phase of CyIIa expression in the gut, shares homologies with the late module of the endoderm-specific Endo16 gene (endo16 Module B). Oligonucleotides containing replicas of these two target sites fused upstream of a sea urchin basal promoter are sufficient to confer accurate mesenchyme and late gut expression of an injected GFP construct. The finding of a single protein target site that recapitulates CyIIa expression in both primary and secondary mesenchyme cells suggests the existence of a pan-mesodermal gene expression program in the sea urchin embryo.
Assuntos
Actinas/genética , Actinas/fisiologia , Regulação da Expressão Gênica no Desenvolvimento , Mesoderma/metabolismo , Transcrição Gênica , Animais , Sequência de Bases , Sítios de Ligação , Ligação Competitiva , Evolução Biológica , Núcleo Celular/metabolismo , DNA/metabolismo , Gástrula/metabolismo , Genes Reporter , Proteínas de Fluorescência Verde , Proteínas Luminescentes/metabolismo , Modelos Genéticos , Dados de Sequência Molecular , Ligação Proteica , Proteínas Recombinantes de Fusão/metabolismo , Ouriços-do-Mar , Homologia de Sequência do Ácido Nucleico , Fatores de Transcrição/metabolismoRESUMO
Strongylocentrotus purpuratus Otx (SpOtx) is required simultaneously in sea urchin development for the activation of endo16 in the vegetal plate and for the activation of spec2a in the aboral ectoderm. Because Otx binding sites alone do not appear to be responsible for the spatially restricted expression of spec2a, additional DNA elements were sought. We show here that consensus Otx binding sites fused to basal promoters are sufficient to activate CAT reporter gene expression in all cell types, although expression in endomesoderm progenitors is enhanced. On the other hand, three non-Otx elements derived from the spec2a enhancer are needed together with Otx sites for specifically aboral ectoderm expression. A DNA element termed Y/CBF, lying just downstream from an Otx site within the spec2a enhancer, mediates general activation in the ectoderm. A second element lying between the Otx and Y/CBF sites, called OER, functions to prevent expression in the oral ectoderm. A third site, called ENR, overlapping another Otx site, is required to repress endoderm expression. Three distinct DNA binding proteins interact sequence specifically at the Y/CBF, OER, and ENR elements. The spec2a enhancer thus consists of closely linked activator and repressor elements that function collectively to cause expression of the spec2a gene in the aboral ectoderm.
Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Homeodomínio/genética , Ouriços-do-Mar/embriologia , Ouriços-do-Mar/genética , Animais , Sequência de Bases , Sítios de Ligação/genética , Padronização Corporal/genética , Primers do DNA/genética , Elementos Facilitadores Genéticos , Genes Reguladores , Genes Reporter , Dados de Sequência Molecular , Fatores de Transcrição Otx , Regiões Promotoras GenéticasRESUMO
In embryos of indirectly developing echinoids, the secondary (oral-aboral) larval axis is established after fertilization by an as yet undiscovered process. One of the earliest manifestations of this axis is an asymmetry in mitochondrial respiration, with the prospective oral side of the embryo exhibiting a higher rate of respiration than the prospective aboral side. We show here that respiratory asymmetry can be experimentally induced within embryos by immobilizing them in tight clusters of four ("rosettes"). Within such clusters a redox gradient is established from the inside to the outside of the rosette. Vital staining of clustered embryos demonstrates that the side of the embryo facing the outside of the rosette (i.e., the most oxidizing) tends to become the oral side, while the side facing the inside tends to become the aboral side. Effective entrainment of the oral-aboral axis requires that the embryos remain immobilized in rosettes until the hatching blastula stage. To begin to investigate the molecular mechanisms underlying this effect we made use of P3A2, a transcriptional regulatory protein whose activity is spatially modulated along the oral-aboral axis. When synthetic mRNA encoding P3A2 fused to the VP16 activation domain is injected into eggs, it activates embryonic expression of a green fluorescent protein reporter gene containing a basal promoter and a single strong P3A2 target site. In embryo rosettes, such activation occurs predominantly on the outside of the rosette, suggesting that the activity of the P3A2 protein is spatially regulated by the respiratory asymmetry established by clustering the embryos. These findings are discussed with reference to earlier work on both oral-aboral axis specification and P3A2 and used to develop a testable model of the mechanism of oral-aboral axis specification in the sea urchin embryo.
Assuntos
Ouriços-do-Mar/embriologia , Animais , Sequência de Bases , Padronização Corporal , Fase de Clivagem do Zigoto/citologia , Fase de Clivagem do Zigoto/metabolismo , Primers do DNA/genética , Proteínas de Ligação a DNA/genética , Feminino , Genes Reporter , Proteínas de Fluorescência Verde , Proteínas Luminescentes/genética , Masculino , Mitocôndrias/metabolismo , Modelos Biológicos , Oxirredução , Consumo de Oxigênio , Ouriços-do-Mar/genética , Ouriços-do-Mar/metabolismoRESUMO
The endo16 gene of Strongylocentrotus purpuratus encodes a secreted protein of the embryonic and larval midgut. The overall functional organization of the spatial and temporal control system of this gene are relatively well known from a series of earlier cis-regulatory studies. Our recent computational model for the logic operations of the proximal region of the endo16 control system (Module A) specifies the function of interactions at each transcription factor target site of Module A. Here, we extend sequence level functional analysis to the adjacent cis-regulatory region, Module B. The computational logic model is broadened to include B/A interactions as well as other Module B functions. Module B drives expression later in development and its major activator is responsible for a sharp, gut-specific increase in transcription after gastrulation. As shown earlier, Module B output undergoes a synergistic amplification that requires interactions within Module A. The interactions within Module B that are required to generate and transmit its output to Module A are identified. Logic considerations predicted an internal cis-regulatory switch by which spatial control of endo16 expression is shifted from Module A (early) to Module B (later). This prediction was confirmed experimentally and a distinct set of interactions in Module B that mediate the switch function was demonstrated. The endo16 computational model now provides a detailed explanation of the information processing functions executed by the cis-regulatory system of this gene throughout embryogenesis. Early in development the gene participates in the specification events that define the endomesoderm; later it functions as a gut-specific differentiation gene. The cis-regulatory switch mediates this functional change.
Assuntos
Moléculas de Adesão Celular/genética , DNA/genética , Regulação da Expressão Gênica no Desenvolvimento , Regiões Promotoras Genéticas/genética , Proteínas/genética , Ouriços-do-Mar/genética , Animais , Sequência de Bases , DNA/metabolismo , Embrião não Mamífero/metabolismo , Genes Reporter , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Hibridização In Situ , Cinética , Modelos Biológicos , Dados de Sequência Molecular , Fatores de Transcrição Otx , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Sequências Reguladoras de Ácido Nucleico , Ouriços-do-Mar/embriologia , Ouriços-do-Mar/metabolismo , Transcrição Gênica/genéticaRESUMO
New technologies for isolating differentially expressed genes from large arrayed cDNA libraries are reported. These methods can be used to identify genes that lie downstream of developmentally important transcription factors and genes that are expressed in specific tissues, processes, or stages of embryonic development. Though developed for the study of gene expression during the early embryogenesis of the sea urchin Strongylocentrotus purpuratus, these technologies can be applied generally. Hybridization parameters were determined for the reaction of complex cDNA probes to cDNA libraries carried on six nylon filters, each containing duplicate spots from 18,432 bacterial clones (macroarrays). These libraries are of sufficient size to include nearly all genes expressed in the embryo. The screening strategy we have devised is designed to overcome inherent sensitivity limitations of macroarray hybridization and thus to isolate differentially expressed genes that are represented only by low-prevalence mRNAs. To this end, we have developed improved methods for the amplification of cDNA from small amounts of tissue (as little as approximately 300 sea urchin embryos, or 2 x 10(5) cells, or about 10 ng of mRNA) and for the differential enhancement of probe sequence concentration by subtractive hybridization. Quantitative analysis of macroarray hybridization shows that these probes now suffice for detection of differentially expressed mRNAs down to a level below five molecules per average embryo cell.
Assuntos
Embrião não Mamífero/fisiologia , Regulação da Expressão Gênica no Desenvolvimento , Biblioteca Gênica , Análise de Sequência com Séries de Oligonucleotídeos , Ouriços-do-Mar/embriologia , Ouriços-do-Mar/genética , Animais , Animais Geneticamente Modificados , Sondas de DNA , DNA Complementar , Genes Reporter , Proteínas de Fluorescência Verde , Proteínas Luminescentes/análise , Proteínas Luminescentes/genética , RNA Mensageiro/genética , Fatores de Transcrição/metabolismoRESUMO
The Hox cluster of the sea urchin Strongylocentrous purpuratus contains ten genes in a 500 kb span of the genome. Only two of these genes are expressed during embryogenesis, while all of eight genes tested are expressed during development of the adult body plan in the larval stage. We report the spatial expression during larval development of the five 'posterior' genes of the cluster: SpHox7, SpHox8, SpHox9/10, SpHox11/13a and SpHox11/13b. The five genes exhibit a dynamic, largely mesodermal program of expression. Only SpHox7 displays extensive expression within the pentameral rudiment itself. A spatially sequential and colinear arrangement of expression domains is found in the somatocoels, the paired posterior mesodermal structures that will become the adult perivisceral coeloms. No such sequential expression pattern is observed in endodermal, epidermal or neural tissues of either the larva or the presumptive juvenile sea urchin. The spatial expression patterns of the Hox genes illuminate the evolutionary process by which the pentameral echinoderm body plan emerged from a bilateral ancestor.
Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Genes Homeobox , Mesoderma/fisiologia , Família Multigênica , Ouriços-do-Mar/crescimento & desenvolvimento , Ouriços-do-Mar/genética , Animais , Evolução Biológica , Padronização Corporal , Larva , Ouriços-do-Mar/citologiaRESUMO
Results of a first-stage Sea Urchin Genome Project are summarized here. The species chosen was Strongylocentrotus purpuratus, a research model of major importance in developmental and molecular biology. A virtual map of the genome was constructed by sequencing the ends of 76,020 bacterial artificial chromosome (BAC) recombinants (average length, 125 kb). The BAC-end sequence tag connectors (STCs) occur an average of 10 kb apart, and, together with restriction digest patterns recorded for the same BAC clones, they provide immediate access to contigs of several hundred kilobases surrounding any gene of interest. The STCs survey >5% of the genome and provide the estimate that this genome contains approximately 27,350 protein-coding genes. The frequency distribution and canonical sequences of all middle and highly repetitive sequence families in the genome were obtained from the STCs as well. The 500-kb Hox gene complex of this species is being sequenced in its entirety. In addition, arrayed cDNA libraries of >10(5) clones each were constructed from every major stage of embryogenesis, several individual cell types, and adult tissues and are available to the community. The accumulated STC data and an expanding expressed sequence tag database (at present including >12, 000 sequences) have been reported to GenBank and are accessible on public web sites.
Assuntos
Genoma , Mapeamento Físico do Cromossomo , Ouriços-do-Mar/genética , Envelhecimento/genética , Animais , Clonagem Molecular , Mapeamento de Sequências Contíguas , DNA Complementar/genética , Bases de Dados Factuais , Etiquetas de Sequências Expressas , Regulação da Expressão Gênica no Desenvolvimento , Genes Homeobox/genética , Internet , Dados de Sequência Molecular , Família Multigênica/genética , Análise de Sequência com Séries de Oligonucleotídeos , Sequências Repetitivas de Ácido Nucleico/genética , Ouriços-do-Mar/citologia , Ouriços-do-Mar/embriologiaRESUMO
The adult body plan of bilaterians is achieved by imposing regional specifications on pluripotential cells. The establishment of spatial domains is governed in part by regulating expression of transcription factors. The key to understanding bilaterian evolution is contingent on our understanding of how the regulation of these transcription factors influenced bilaterian stem-group evolution.
Assuntos
Evolução Biológica , Padronização Corporal/genética , Invertebrados/classificação , Invertebrados/genética , Animais , Embrião não Mamífero , Regulação da Expressão Gênica , Genes Homeobox , Filogenia , Fatores de Transcrição/genéticaRESUMO
Putative fossil embryos and larvae from the Precambrian phosphorite rocks of the Doushantuo Formation in Southwest China have been examined in thin section by bright field and polarized light microscopy. Although we cannot completely exclude a nonbiological or nonmetazoan origin, we identified what appear to be modern cnidarian developmental stages, including both anthozoan planula larvae and hydrozoan embryos. Most importantly, the sections contain a variety of small (=200 microm) structures that greatly resemble gastrula stage embryos of modern bilaterian forms.
Assuntos
Evolução Biológica , Cnidários/embriologia , Embrião não Mamífero/anatomia & histologia , Fósseis , Variação Genética , Animais , Padronização Corporal , China , Gástrula , Paleontologia/métodosRESUMO
A prediction from the set-aside theory of bilaterian origins is that pattern formation processes such as those controlled by the Hox cluster genes are required specifically for adult body plan formation. This prediction can be tested in animals that use maximal indirect development, in which the embryonic formation of the larva and the postembryonic formation of the adult body plan are temporally and spatially distinct. To this end, we quantitatively measured the amount of transcripts for five Hox genes in embryos of a lophotrochozoan, the polychaete annelid Chaetopterus sp. The polychaete Hox complex is shown not to be expressed during embryogenesis, but transcripts of all measured Hox complex genes are detected at significant levels during the initial stages of adult body plan formation. Temporal colinearity in the sequence of their activation is observed, so that activation follows the 3'-5' arrangement of the genes. Moreover, Hox gene expression is spatially localized to the region of teloblastic set-aside cells of the later-stage embryos. This study shows that an indirectly developing lophotrochozoan shares with an indirectly developing deuterostome, the sea urchin, a common mode of Hox complex utilization: construction of the larva, whether a trochophore or dipleurula, does not involve Hox cluster expression, but in both forms the complex is expressed in the set-aside cells from which the adult body plan derives.
Assuntos
Evolução Biológica , Proteínas Fetais , Regulação da Expressão Gênica no Desenvolvimento , Genes Homeobox , Família Multigênica , Poliquetos/embriologia , Poliquetos/genética , Animais , Embrião não Mamífero/fisiologia , Evolução Molecular , Feminino , Masculino , Dados de Sequência Molecular , Oócitos/fisiologia , Espermatozoides/fisiologia , Proteínas com Domínio T/genética , Transcrição GênicaRESUMO
Several recent laboratory observations that bear on the origin of the Bilateria are reviewed and interpreted in light of our set-aside cell theory for bilaterian origins. We first discuss new data concerning the phylogeny of bilaterian phyla. Next, we use systematic, molecular, and paleontological lines of evidence to argue that the latest common ancestor of echinoderms plus hemichordates used a maximal indirect mode of development. Furthermore, the latest common ancestor of molluscs and annelids was also indirectly developing. Finally, we discuss new data on Hox gene expression patterns which suggest that both sea urchins and polychaete annelids use Hox genes in a very similar fashion. Neither utilizes the complete Hox complex in the development of the larva per se, while the Hox complex is expressed in the set-aside cells from which the adult body plan is formed. Our current views on the ancestry of the bilaterians are summarized in phylogenetic terms, incorporating the characters discussed in this paper.
Assuntos
Evolução Biológica , Invertebrados/crescimento & desenvolvimento , Invertebrados/genética , Animais , Anelídeos , Cordados não Vertebrados , Equinodermos , Regulação da Expressão Gênica no Desenvolvimento , Genes Homeobox , Modelos Biológicos , Moluscos , Família Multigênica , FilogeniaAssuntos
Proteínas da Matriz Extracelular/genética , Genes , Família Multigênica , Ouriços-do-Mar/genética , Motivos de Aminoácidos/genética , Sequência de Aminoácidos , Animais , Sequência Consenso , DNA Complementar/genética , Proteínas da Matriz Extracelular/química , Biblioteca Gênica , Dados de Sequência Molecular , Ouriços-do-Mar/embriologia , Ouriços-do-Mar/ultraestrutura , Homologia de Sequência de AminoácidosRESUMO
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.
Assuntos
Equinodermos/embriologia , Equinodermos/crescimento & desenvolvimento , Fósseis , Animais , Equinodermos/genética , Genes HomeoboxRESUMO
The process of embryogenesis depends on differential regulation of genes in the spatial components defined by the embryonic cells (blastomeres). Developmental regulation is mediated by complex, hardwired genomic control systems consisting of clusters of multiple target sites at which specific interactions with regionally presented transcription factors occur. In the age of genomics and gene-transfer technology, the sea urchin embryo provides unique opportunities for experimental analysis of these processes. Research on gene regulation in sea urchin embryos in the past year has seen remarkable progress in two large areas: in understanding functional cis-regulatory architecture; and in understanding the mechanism by which the axial coordinates of the egg are transduced into a molecular system for differential gene activation.
Assuntos
Ouriços-do-Mar/genética , Transcrição Gênica , Animais , Sequência de Bases , DNA , Genoma , Dados de Sequência Molecular , Ouriços-do-Mar/embriologia , Fatores de Transcrição/fisiologiaRESUMO
A memorable workshop, focused on causal mechanisms in metazoan evolution and sponsored by NASA, was held in early June 1998, at MBL. The workshop was organized by Mike Levine and Eric H. Davidson, and it included the PI and associates from 12 different laboratories, a total of about 30 people. Each laboratory had about two and one half hours in which to represent its recent research and cast up its current ideas for an often intense discussion. In the following we have tried to enunciate some of the major themes that emerged, and to reflect on their implications. The opinions voiced are our own. We would like to tender apologies over those contributions we have not been able to include, but this is not, strictly speaking, a meeting review. Rather we have focused on those topics that bear more directly on evolutionary mechanisms, and have therefore slighted some presentations (including some of our own), that were oriented mainly toward developmental processes. J. Exp. Zool. (Mol. Dev. Evol. ) 285:104-115, 1999.
Assuntos
Evolução Biológica , Regulação da Expressão Gênica , Genes Homeobox , Animais , Padronização Corporal , Caenorhabditis elegans/genética , Evolução Molecular , Filogenia , Estados Unidos , United States National Aeronautics and Space AdministrationRESUMO
A set of 956 expressed sequence tags derived from 7-hour (mid-cleavage) sea urchin embryos was analyzed to assess biosynthetic functions and to illuminate the structure of the message population at this stage. About a quarter of the expressed sequence tags represented repetitive sequence transcripts typical of early embryos, or ribosomal and mitochondrial RNAs, while a majority of the remainder contained significant open reading frames. A total of 232 sequences, including 153 different proteins, produced significant matches when compared against GenBank. The majority of these identified sequences represented 'housekeeping' proteins, i.e., cytoskeletal proteins, metabolic enzymes, transporters and proteins involved in cell division. The most interesting finds were components of signaling systems and transcription factors not previously reported in early sea urchin embryos, including components of Notch and TGF signal transduction pathways. As expected from earlier kinetic analyses of the embryo mRNA populations, no very prevalent protein-coding species were encountered; the most highly represented such sequences were cDNAs encoding cyclins A and B. The frequency of occurrence of all sequences within the database was used to construct a sequence prevalence distribution. The result, confirming earlier mRNA population analyses, indicated that the poly(A) RNA of the early embryo consists mainly of a very complex set of low-copy-number transcripts.