Studying Tunicata WBR Using Botrylloides anceps

Abstract

Tunicates are marine filter-feeding invertebrates that can be found worldwide and which are the closest phylogenetic group to the vertebrates (Craniata). Of particular interest, colonial tunicates are the only known chordates that can undergo Whole-Body Regeneration (WBR) via vascular budding. In Botrylloides anceps, a fully functional adult regenerates from a fragment of the vascular system in around 2 weeks after amputation. In this chapter, we present protocols to collect B. anceps colonies, confirm their species, breed them in the lab, monitor WBR and perform histological staining on cryosections.

Phylogeographic and morphological analysis ofBotrylloides nigerHerdman, 1886 from the northeastern Mediterranean Sea

“…Spatial samples were collected from the coastal areas of Antalya (Kemer-Side--Alanya-Tersane), Mersin (Tisan-Kızkalesi-Mezitli), and Hatay (Konacık) in September and October 2018) in water depth between 30-50 cm as previously described for Botrylloides anceps (Figure 1A) (Karahan et al, 2022). Samples were collected from the submerged stones using a single-edged razor blade.…”

“…DNA extractions were completed based on the protocol of (Karahan et al, 2022). The amplification reactions were executed based on Reem et al (2018) for COI, H3, 18S, and 28S genes (Table S4).…”

“…The blastogenic cycle culminates with the death and absorption of all parental zooids by the colony and the emergence of a new generation of zooids in a process known as the takeover (Manni et al, 2019). Natural chimerism following the fusion of two closely related kins, hibernation and even whole-body regeneration have commonly but not exhaustively been reported in Botrylloides (Paz & Rinkevich, 2002; Rinkevich, 2005; Sheets et al, 2016; Zondag et al, 2016; Blanchoud et al, 2017; Blanchoud et al, 2018a; Karahan et al, 2022). The Botrylloides genus is composed of 21 reported species, all of which are morphologically very close with zooids aligned in a ladder-like arrangement.…”

Variation in whole-body regeneration between Botrylloides morphs and species

“…In contrast to the WBR of B. diegensis , multiple zooids are formed in the WBR of B. violaceus . Recently, B. anceps was reported to orchestrate single zooid WBR via stem-cell niche formation, while there is no previous record on B. jacksonianum (Karahan et al, 2022).…”

Identification and characterization ofBotrylloides(Styelidae) species from Aotearoa New Zealand coasts

“…WBR, blastogenesis, hibernation/aestivation, and natural chimerism are reported only within the Styelidae ascidians. To date, examples of WBR have been described for Botrylloides violaceus (Oka, 1927) , Botrylloides diegensis (Ritter & Forsyth, 1917), Botryllus primigenus (Oka, 1928) , Botrylloides anceps (Herdman, 1891) and Botryllus schlosseri , Pallas (1766) (Blanchoud, Rinkevich, et al, 2018; Karahan et al, 2022; Rosner et al, 2019). B. diegensis can regenerate continuously from a little vascular tissue that includes at least ~200 cells in as little as 10 days (Blanchoud, Rutherford, et al, 2018; Rinkevich et al, 1995).…”