Whole body regeneration and developmental competition in two botryllid ascidians.

BACKGROUND: Botryllid ascidians are a group of marine invertebrate chordates that are colonial and grow by repeated rounds of asexual reproduction to form a colony of individual bodies, called zooids, linked by a common vascular network. Two distinct processes are responsible for zooid regeneration. In the first, called blastogenesis, new zooids arise from a region of multipotent epithelium from a pre-existing zooid. In the second, called whole body regeneration (WBR), mobile cells in the vasculature coalesce and are the source of the new zooid. In some botryllid species, blastogenesis and WBR occur concurrently, while in others, blastogenesis is used exclusively for growth, while WBR only occurs following injury or exiting periods of dormancy. In species such as Botrylloides diegensis, injury induced WBR is triggered by the surgical isolation of a small piece of vasculature. However, Botryllus schlosseri has unique requirements that must be met for successful injury induced WBR. Our goal was to understand why there would be different requirements between these two species. RESULTS: While WBR in B. diegensis was robust, we found that in B. schlosseri, new zooid growth following injury is unlikely due to circulatory cells, but instead a result of ectopic development of tissues leftover from the blastogenic process. These tissues could be whole, damaged, or partially resorbed developing zooids, and we defined the minimal amount of vascular biomass to support ectopic regeneration. We did find a common theme between the two species: a competitive process exists which results in only a single zooid reaching maturity following injury. We utilized this phenomenon and found that competition is reversible and mediated by circulating factors and/or cells. CONCLUSIONS: We propose that WBR does not occur in B. schlosseri and that the unique requirements defined in other studies only serve to increase the chances of ectopic development. This is likely a response to injury as we have discovered a vascular-based reversible competitive mechanism which ensures that only a single zooid completes development. This competition has been described in other species, but the unique response of B. schlosseri to injury provides a new model to study resource allocation and competition within an individual.

The biology of the extracorporeal vasculature of Botryllus schlosseri.

The extracorporeal vasculature of the colonial ascidian Botryllus schlosseri plays a key role in several biological processes: transporting blood, angiogenesis, regeneration, self-nonself recognition, and parabiosis. The vasculature also interconnects all individuals in a colony and is composed of a single layer of ectodermally-derived cells. These cells form a tube with the basal lamina facing the lumen, and the apical side facing an extracellular matrix that consists of cellulose and other proteins, known as the tunic. Vascular tissue is transparent and can cover several square centimeters, which is much larger than any single individual within the colony. It forms a network that ramifies and expands to the perimeter of each colony and terminates into oval-shaped protrusions known as ampullae. Botryllus individuals replace themselves through a weekly budding cycle, and vasculature is added to ensure the interconnection of each new individual, thus there is continuous angiogenesis occurring naturally. The vascular tissue itself is highly regenerative; surgical removal of the ampullae and peripheral vasculature triggers regrowth within 24-48 h, which includes forming new ampullae. When two individuals, whether in the wild or in the lab, come into close contact and their ampullae touch, they can either undergo parabiosis through anastomosing vessels, or reject vascular fusion. The vasculature is easily manipulated by direct means such as microinjections, microsurgeries, and pharmacological reagents. Its transparent nature allows for in vivo analysis by bright field and fluorescence microscopy. Here we review the techniques and approaches developed to study the different biological processes that involve the extracorporeal vasculature.

Cellular and molecular mechanisms of regeneration in colonial and solitary Ascidians.

Regenerative ability is highly variable among the metazoans. While many invertebrate organisms are capable of complete regeneration of entire bodies and organs, whole-organ regeneration is limited to very few species in the vertebrate lineages. Tunicates, which are invertebrate chordates and the closest extant relatives of the vertebrates, show robust regenerative ability. Colonial ascidians of the family of the Styelidae, such as several species of Botrylloides, are able to regenerate entire new bodies from nothing but fragments of vasculature, and they are the only chordates that are capable of whole body regeneration. The cell types and signaling pathways involved in whole body regeneration are not well understood, but some evidence suggests that blood borne cells may play a role. Solitary ascidians such as Ciona can regenerate the oral siphon and their central nervous system, and stem cells located in the branchial sac are required for this regeneration. Here, we summarize the cellular and molecular mechanisms of tunicate regeneration that have been identified so far and discuss differences and similarities between these mechanisms in regenerating tunicate species.