The architecture of cell differentiation in choanoflagellates and sponge choanocytes

Abstract

Although collar cells are conserved across animals and their closest relatives, the choanoflagellates, little is known about their ancestry, their subcellular architecture, or how they differentiate. The choanoflagellate Salpingoeca rosetta expresses genes necessary for animal development and can alternate between unicellular and multicellular states, making it a powerful model for investigating the origin of animal multicellularity and mechanisms underlying cell differentiation. To compare the subcellular architecture of solitary collar cells in S. rosetta with that of multicellular ‘rosette’ colonies and collar cells in sponges, we reconstructed entire cells in 3D through transmission electron microscopy on serial ultrathin sections. Structural analysis of our 3D reconstructions revealed important differences between single and colonial choanoflagellate cells, with colonial cells exhibiting a more amoeboid morphology consistent with higher levels of macropinocytotic activity. Comparison of multiple reconstructed rosette colonies highlighted the variable nature of cell sizes, cell–cell contact networks, and colony arrangement. Importantly, we uncovered the presence of elongated cells in some rosette colonies that likely represent a distinct and differentiated cell type, pointing toward spatial cell differentiation. Intercellular bridges within choanoflagellate colonies displayed a variety of morphologies and connected some but not all neighbouring cells. Reconstruction of sponge choanocytes revealed ultrastructural commonalities but also differences in major organelle composition in comparison to choanoflagellates. Together, our comparative reconstructions uncover the architecture of cell differentiation in choanoflagellates and sponge choanocytes and constitute an important step in reconstructing the cell biology of the last common ancestor of animals.

Abstract

3D electron microscopy of choanoflagellates and sponge choanocytes reveals a remarkable variety of cell architecture and suggests that cell type differentiation may have been present in the stem lineage leading to the animals.

Author summary

Choanoflagellates are microscopic aquatic organisms that can alternate between single-celled and multicellular states, and sequencing of their genomes has revealed that choanoflagellates are the closest single-celled relatives of animals. Moreover, choanoflagellates are a form of ‘collar cell’—a cell type crowned by an array of finger-like microvilli and a single, whip-like flagellum. This cell type is also found throughout the animal kingdom; therefore, studying the structure of the choanoflagellate collar cell can shed light on how this cell type and animal multicellularity might have evolved. We used electron microscopy to reconstruct in 3D the total subcellular composition of single-celled and multicellular choanoflagellates as well as the collar cells from a marine sponge, which represents an early-branching animal lineage. We found differences between single-celled and multicellular choanoflagellates in structures associated with cellular energetics, membrane trafficking, and cell morphology. Likewise, we describe a complex system of cell–cell connections associated with multicellular choanoflagellates. Finally, comparison of choanoflagellates and sponge collar cells revealed subcellular differences associated with feeding and cellular energetics. Taken together, this study is an important step forward in reconstructing the biology of the last common ancestor of the animals.