Distal regulation, silencers and a shared combinatorial syntax are hallmarks of animal embryogenesis

The chromatin environment plays a central role in regulating developmental gene expression in metazoans. Yet, the basal regulatory landscape of metazoan embryogenesis is unknown. Here, we generate chromatin accessibility profiles for six embryonic, plus larval and adult stages in the sponge Amphimedon queenslandica. These profiles are reproducible within stages, reflect histone modifications, and identify transcription factor (TF) binding sequence motifs predictive of cis-regulatory elements during embryogenesis in other metazoans but not the unicellular relative Capsaspora. Motif analysis of chromatin accessibility profiles across Amphimedon embryogenesis identifies three major developmental periods. As in bilaterian embryogenesis, early development in Amphimedon involves activating and repressive chromatin in regions both proximal and distal to transcription start sites. Transcriptionally repressive elements (silencers) are prominent during late embryogenesis and coincide with an increase in cis-regulatory regions harbouring metazoan TF binding motifs, and an increase in the expression of metazoan-specific genes. Changes in chromatin state and gene expression in Amphimedon suggest the conservation of distal enhancers, dynamically silenced chromatin, and TF-DNA binding specificity in animal embryogenesis.

The buds of Oscarella lobularis (Porifera): a new convenient model for sponge cell and developmental biology

The comparative study of the four non-bilaterian phyla (Cnidaria, Placozoa, Ctenophora, Porifera) should provide insights into the origin of bilaterian traits. Except for Cnidaria, present knowledge on the cell biology and development of these animals is so far limited. Non-bilaterian models are needed to get further into cell architecture and molecular mechanisms.Given the developmental, histological, ecological and genomic differences between the four sponge classes, we develop a new sponge model: the buds of the Oscarella lobularis (class Homoscleromorpha). This experimental model supplements the two other most famous sponge models Amphimedon queenslandica and Ephydatia muelleri, both belonging to the class Demospongiae.Budding is a natural and spontaneous asexual reproduction mean, but budding can be triggered in vitro ensuring availability of biological material all year long. We provide a full description of buds, from their formation to their development into juveniles. Their transparency enables fluorescent and live imaging, and their abundance allows for experimental replicates. Moreover, regeneration and cell reaggregation capabilities provide interesting experimental morphogenetic contexts. The numerous techniques now mastered on these buds make it a new suitable sponge model.Summary statementStudying sponge biology is needed to understand the evolution of metazoans. We developed a new model suitable for experimental biology that allows to study morphogenetic processes with modern tools.

Are there systematic biases in RNA-seq data analysis? A case study for Amphimedon queenslandica sponge as a model object

BACKGROUNDThe performance of a functional annotation approach for RNA-seq bioinformatics pipelines was to be compared with the method where groups of genes are generated with no relation to ontologes. Three publicly available RNA-Seq experiments for Amphimedon queenslandica sponge were used for the designed comparison. One of these experiments was referred in the publication where stages of embryo development were compared for a wide range of animal species.METHODSThe expression levels were re-calculated here for three independent series of experiments. The functional annotation of differential expression levels was than conducted. This allow to compare an applicability of the two approaches, and to re-evaluate the interpretation provided in the mentioned publication.RESULTSIt was confirmed by the conventional approach that Wnt and Notch pathways do operate in a development of a sponge embryo. The method of annotation which uses unbounded grouping of genes was effective in an ability to separate development stages of sponge embryo. In addition, the published results were by a suggestion distorted by an artifact, caused by a positive feedback in the stage of data processing.

Co-expression of synaptic genes in the sponge Amphimedon queenslandica uncovers ancient neural submodules

The synapse is a complex cellular module crucial to the functioning of neurons. It evolved largely through the exaptation of pre-existing smaller submodules, each of which are comprised of ancient sets of proteins that are conserved in modern animals and other eukaryotes. Although these ancient submodules themselves have non-neural roles, it has been hypothesized that they may mediate environmental sensing behaviors in aneural animals, such as sponges. Here we identify orthologues in the sponge Amphimedon queenslandica of genes encoding synaptic submodules in neural animals, and analyse their cell-type specific and developmental expression to determine their potential to be co-regulated. We find that genes comprising certain synaptic submodules, including those involved in vesicle trafficking, calcium-regulation and scaffolding of postsynaptic receptor clusters, are co-expressed in adult choanocytes and during metamorphosis. Although these submodules may contribute to sensory roles in this cell type and this life cycle stage, total synaptic gene co-expression profiles do not support the existence of a functional synapse in A. queenslandica. The lack of evidence for the co-regulation of genes necessary for pre- and post-synaptic functioning in A. queenslandica suggests that sponges, and perhaps the last common ancestor of sponges and other extant animals, had the ability to promulgate sensory inputs without complete synapse-like functionalities. The differential co-expression of multiple synaptic submodule genes in sponge choanocytes, which have sensory and feeding roles, however, is consistent with the metazoan ancestor minimally being able to undergo exo- and endocytosis in a controlled and localized manner.

Early origin and deep conservation of enhancers in animals

Transcription factors (TFs) bind DNA enhancer sequences to regulate gene transcription in animals. Unlike TFs, the evolution of enhancers has been difficult to trace because of their rapid evolution. Here, we show enhancers from the sponge Amphimedon queenslandica can drive cell type-specific reporter gene expression in zebrafish and mouse, despite sponge and vertebrate lineages diverging over 700 million years ago. Although sponge enhancers, which are present in both highly conserved syntenic gene regions (Islet–Scaper, Ccne1–Uri and Tdrd3–Diaph3) and sponge-specific intergenic regions, have no significant sequence identity with vertebrate genomic sequences, the type and frequency of TF binding motifs in the sponge enhancer allow for the identification of homologous enhancers in bilaterians. Islet enhancers identified in human and mouse Scaper genes drive zebrafish reporter expression patterns that are almost identical to the sponge Islet enhancer. The existence of homologous enhancers in these disparate metazoans suggests animal development is controlled by TF-enhancer DNA interactions that were present in the first multicellular animals.One-sentence summaryEnhancer activity is conserved across 700 million years of trans-phyletic divergence.