Abstract
The colonial marine invertebrate Botrylloides leachii belongs to the Tunicata subphylum, the closest invertebrate relatives to the vertebrate group and the only known class of chordates that can undergo whole-body regeneration (WBR). This dramatic developmental process allows a minute isolated fragment of B. leachii’s vascular system, or a colony excised of all adults, to restore a functional animal in as little as 10 days. In addition to this exceptional regenerative capacity, B. leachii can reproduce both sexually, through a tadpole larval stage, and asexually, through palleal budding. Thus, three alternative developmental strategies lead to the establishment of filter-feeding adults. Consequently, B. leachii is particularly well suited for comparative studies on regeneration and should provide novel insights into regenerative processes in chordates.
Here, after a short introduction on regeneration, we overview the biology of B. leachii as well as the current state of knowledge on WBR in this species and in related species of tunicates. Finally, we highlight the possible future directions that research might take in the study of WBR, including thoughts on technological approaches that appear most promising in this context. Overall, we provide a synthesis of the current knowledge on WBR in B. leachii to support research in this chordate species.
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References
Bely AE, Nyberg KG (2010) Evolution of animal regeneration: re-emergence of a field. Trends Ecol Evol 25:161–170. https://doi.org/10.1016/j.tree.2009.08.005
Berna L, Alvarez-Valin F (2014) Evolutionary genomics of fast evolving tunicates. Genome Biol Evol 6:1724–1738. https://doi.org/10.1093/gbe/evu122
Berrill NJ (1947) The developmental cycle of Botrylloides. Q J Microsc Sci 88:393–407
Berrill NJ, Cohen A (1936) Regeneration in Clavelina lepadiformis. J Exp Biol 13:352–362
Blanchoud S, Zondag L, Lamare MD, Wilson MJ (2017) Hematological analysis of the ascidian Botrylloides leachii (Savigny, 1816) during whole-body regeneration. Biol Bull 232:143–157. https://doi.org/10.1086/692841
Brewin BI (1946) Ascidians in the vicinity of the Portobello Marine Biological Station, Otago Harbour. Trans R Soc N Z 76:87–131
Brockes JP, Kumar A, Velloso CP (2001) Regeneration as an evolutionary variable. J Anat 199:3–11. https://doi.org/10.1017/S0021878201008299
Brown FD, Swalla BJ (2012) Evolution and development of budding by stem cells: ascidian coloniality as a case study. Dev Biol 369:151–162. https://doi.org/10.1016/j.ydbio.2012.05.038
Brown FD, Keeling EL, Le AD, Swalla BJ (2009) Whole body regeneration in a colonial ascidian, Botrylloides violaceus. J Exp Zool B Mol Dev Evol 312:885–900. https://doi.org/10.1002/jez.b.21303
Brunetti R (1976) Biological cycle of Botrylloides leachi (Savigny) (Ascidiacea) in the Venetian lagoon. Vie Milieu XXVI:105–122
Burighel P, Brunetti R, Zaniolo G (1976) Hibernation of the colonial ascidian Botrylloides leachi (Savigny): histological observations. Ital J Zool 43:293–301. https://doi.org/10.1080/11250007609430146
Chambon J-P, Nakayama A, Takamura K et al (2007) ERK- and JNK-signalling regulate gene networks that stimulate metamorphosis and apoptosis in tail tissues of ascidian tadpoles. Development 134:1203–1219. https://doi.org/10.1242/dev.002220
Cima F, Perin A, Burighel P, Ballarin L (2002) Morpho-functional characterization of haemocytes of the compound ascidian Botrylloides leachi (Tunicata, Ascidiacea). Acta Zool 82:261–274. https://doi.org/10.1046/j.1463-6395.2001.00087.x
Davidson B, Swalla BJ (2002) A molecular analysis of ascidian metamorphosis reveals activation of an innate immune response. Development 129:4739–4751
Delsuc F, Brinkmann H, Chourrout D, Philippe H (2006) Tunicates and not cephalochordates are the closest living relatives of vertebrates. Nature 439:965–968. https://doi.org/10.1038/nature04336
Franchi N, Ballin F, Manni L et al (2016) Recurrent phagocytosis-induced apoptosis in the cyclical generation change of the compound ascidian Botryllus schlosseri. Dev Comp Immunol 62:8–16. https://doi.org/10.1016/j.dci.2016.04.011
Franchi N, Ballin F, Ballarin L (2017) Protection from oxidative stress in immunocytes of the colonial ascidian Botryllus schlosseri: transcript characterization and expression studies. Biol Bull 232:45–57. https://doi.org/10.1086/691694
Freeman G (1964) The role of blood cells in the process of asexual reproduction in the tunicate Perophora viridis. J Exp Zool 156:157–183. https://doi.org/10.1002/jez.1401560204
Galliot B (2013) Regeneration in hydra. In: eLS. Wiley, Chichester
Gasparini F, Burighel P, Manni L, Zaniolo G (2008) Vascular regeneration and angiogenic-like sprouting mechanism in a compound ascidian is similar to vertebrates. Evol Dev 10:591–605. https://doi.org/10.1111/j.1525-142X.2008.00274.x
Gasparini F, Caicci F, Rigon F et al (2014) Testing an unusual in vivo vessel network model: a method to study angiogenesis in the colonial tunicate Botryllus schlosseri. Sci Rep 4:6460. https://doi.org/10.1038/srep06460
Gutierrez S, Brown FD (2017) Vascular budding in Symplegma brakenhielmi and the evolution of coloniality in styelid ascidians. Dev Biol 423:152–169. https://doi.org/10.1016/j.ydbio.2017.01.012
Jaźwińska A, Sallin P (2016) Regeneration versus scarring in vertebrate appendages and heart. J Pathol 238:233–246. https://doi.org/10.1002/path.4644
Kassmer SH, Rodriguez D, De Tomaso AW (2016) Colonial ascidians as model organisms for the study of germ cells, fertility, whole body regeneration, vascular biology and aging. Curr Opin Genet Dev 39:101–106. https://doi.org/10.1016/j.gde.2016.06.001
Kawamura K, Fujiwara S (1994) Transdifferentiation of pigmented multipotent epithelium during morphallactic development of budding tunicates. Int J Dev Biol 38:369–377
Kawamura K, Fujiwara S (1995) Cellular and molecular characterization of transdifferentiation in the process of morphallaxis of budding tunicates. Semin Cell Dev Biol 6:117–126. https://doi.org/10.1006/scel.1995.0017
Kawamura K, Hayata D, Fujiwara S, Yubisui T (1998) Serine protease inhibitors expressed in the process of budding of tunicates as revealed by EST analysis. J Biochem 124:1004–1012. https://doi.org/10.1093/oxfordjournals.jbchem.a022192
Kawamura K, Kariya Y, Ono Y et al (2006) Molecular collaborations between serpins and trefoil factor promote endodermal cell growth and gastrointestinal differentiation in budding tunicates. Develop Growth Differ 48:309–322. https://doi.org/10.1111/j.1440-169X.2006.00865.x
Kawamura K, Sugino Y, Sunanaga T, Fujiwara S (2008) Multipotent epithelial cells in the process of regeneration and asexual reproduction in colonial tunicates. Develop Growth Differ 50:1–11. https://doi.org/10.1111/j.1440-169X.2007.00972.x
Kawamura K, Tiozzo S, Manni L et al (2011) Germline cell formation and gonad regeneration in solitary and colonial ascidians. Dev Dyn 240:299–308. https://doi.org/10.1002/dvdy.22542
Kawamura K, Yoshida T, Sekida S (2018) Autophagic dedifferentiation induced by cooperation between TOR inhibitor and retinoic acid signals in budding tunicates. Dev Biol 433:384–393. https://doi.org/10.1016/j.ydbio.2017.08.023
Kürn U, Rendulic S, Tiozzo S, Lauzon RJ (2011) Asexual propagation and regeneration in colonial ascidians. Biol Bull 221:43–61. https://doi.org/10.1086/BBLv221n1p43
Laird DJ, Weissman IL (2004) Telomerase maintained in self-renewing tissues during serial regeneration of the urochordate Botryllus schlosseri. Dev Biol 273:185–194. https://doi.org/10.1016/j.ydbio.2004.05.029
Laird DJ, De Tomaso AW, Weissman IL (2005) Stem cells are units of natural selection in a colonial ascidian. Cell 123:1351–1360. https://doi.org/10.1016/j.cell.2005.10.026
Lauzon RJ, Ishizuka KJ, Weissman IL (2002) Cyclical generation and degeneration of organs in a colonial urochordate involves crosstalk between old and new: a model for development and regeneration. Dev Biol 249:333–348. https://doi.org/10.1006/dbio.2002.0772
Lauzon RJ, Brown C, Kerr L, Tiozzo S (2013) Phagocyte dynamics in a highly regenerative urochordate: insights into development and host defense. Dev Biol 374:357–373. https://doi.org/10.1016/j.ydbio.2012.11.006
Manni L, Zaniolo G, Cima F et al (2007) Botryllus schlosseri: a model ascidian for the study of asexual reproduction. Dev Dyn 236:335–352. https://doi.org/10.1002/dvdy.21037
Michaelsen W, Stephenson TA (1934) The ascidians of the Cape Province of South Africa. Trans R Soc South Afr 22:129–163. https://doi.org/10.1080/00359193409519335
Milkman R (1967) Genetic and developmental studies on Botryllus schlosseri. Biol Bull 132:229. https://doi.org/10.2307/1539891
Millar RH (1971) The biology of ascidians. Adv Mar Biol 9:1–100. https://doi.org/10.1016/S0065-2881(08)60341-7
Mukai H (1977) Comparative studies on the structure of reproductive organs of four botryllid ascidians. J Morphol 152:363–379. https://doi.org/10.1002/jmor.1051520307
Mukai H, Sugimoto K, Taneda Y (1978) Comparative studies on the circulatory system of the compound ascidians, Botryllus, Botrylloides and Symplegma. J Morphol 157:49–78. https://doi.org/10.1002/jmor.1051570105
Mukai H, Saito Y, Watanabe H (1987) Viviparous development in Botrylloides (compound ascidians). J Morphol 193:263–276. https://doi.org/10.1002/jmor.1051930305
Murphy K (2012) Innate immunity: the first lines of defense. In: Janeway’s immunobiology. Taylor & Francis, Oxford, pp 37–73
Myers PE (1990) Space versus other limiting resources for a colonial tunicate, Botrylloides leachii (Savigny), on fouling plates. J Exp Mar Bio Ecol 141:47–52. https://doi.org/10.1016/0022-0981(90)90156-7
Nakayama A, Satou Y, Satoh N (2002) Further characterization of genes expressed during Ciona intestinalis metamorphosis. Differentiation 70:429–437. https://doi.org/10.1046/j.1432-0436.2002.700805.x
Ohashi M, Kawamura K, Fujii N et al (1999) A retinoic acid-inducible modular protease in budding ascidians. Dev Biol 214:38–45. https://doi.org/10.1006/dbio.1999.9400
Oka H, Watanabe H (1957) Vascular budding, a new type of budding in Botryllus. Biol Bull 112:225–240. https://doi.org/10.2307/1539200
Oka H, Watanabe H (1959) Vascular budding in Botrylloides. Biol Bull 117:340. https://doi.org/10.2307/1538913
Okuyama M, Saito Y (2001) Studies on Japanese botryllid ascidians. I. A new species of the genus Botryllus from the Izu Islands. Zool Sci 18:261–267. https://doi.org/10.2108/zsj.18.261
Oren M, Douek J, Fishelson Z, Rinkevich B (2007) Identification of immune-relevant genes in histoincompatible rejecting colonies of the tunicate Botryllus schlosseri. Dev Comp Immunol 31:889–902. https://doi.org/10.1016/j.dci.2006.12.009
Paz G, Rinkevich B (2002) Morphological consequences for multi-partner chimerism in Botrylloides, a colonial urochordate. Dev Comp Immunol 26:615–622
Poss KD (2010) Advances in understanding tissue regenerative capacity and mechanisms in animals. Nat Rev Genet 11:710–722. https://doi.org/10.1038/nrg2879
Rabinowitz C, Rinkevich B (2003) Epithelial cell cultures from Botryllus schlosseri palleal buds: accomplishments and challenges. Methods Cell Sci 25:137–148. https://doi.org/10.1007/s11022-004-2087-9
Rabinowitz C, Rinkevich B (2004) In vitro delayed senescence of extirpated buds from zooids of the colonial tunicate Botryllus schlosseri. J Exp Biol 207:1523–1532. https://doi.org/10.1242/jeb.00899
Rabinowitz C, Rinkevich B (2011) De novo emerged stemness signatures in epithelial monolayers developed from extirpated palleal buds. In Vitro Cell Dev Biol Anim 47:26–31. https://doi.org/10.1007/s11626-010-9357-4
Rabinowitz C, Alfassi G, Rinkevich B (2009) Further portrayal of epithelial monolayers emergent de novo from extirpated ascidians palleal buds. In Vitro Cell Dev Biol Anim 45:334–342. https://doi.org/10.1007/s11626-009-9179-4
Rinkevich B, Rinkevich Y (2013) The “Stars and Stripes” metaphor for animal regeneration-Elucidating two fundamental strategies along a continuum. Cells 2:1–18. https://doi.org/10.3390/cells2010001
Rinkevich B, Shapira M (1998) An improved diet for inland broodstock and the establishment of an inbred line from a colonial sea squirt (Ascidiacea). Aquat Living Resour 11:163–171. https://doi.org/10.1016/S0990-7440(98)80113-7
Rinkevich B, Weissman IL (1987) A long-term study on fused subclones in the ascidian Botryllus schlosseri: the resorption phenomenon (Protochordata: Tunicata). J Zool 213:717–733. https://doi.org/10.1111/j.1469-7998.1987.tb03736.x
Rinkevich B, Shlemberg Z, Lilkerlevav T et al (1993) Life-history characteristics of Botrylloides (Tunicata) populations in Akko Bay, Mediterranean coast of Israel. Isr J Zool 39:197–212. https://doi.org/10.1080/00212210.1993.10688712
Rinkevich B, Shlemberg Z, Fishelson L (1995) Whole-body protochordate regeneration from totipotent blood cells. Proc Natl Acad Sci USA 92:7695–7699
Rinkevich Y, Douek J, Haber O et al (2007a) Urochordate whole body regeneration inaugurates a diverse innate immune signaling profile. Dev Biol 312:131–146. https://doi.org/10.1016/j.ydbio.2007.09.005
Rinkevich Y, Paz G, Rinkevich B, Reshef R (2007b) Systemic bud induction and retinoic acid signaling underlie whole body regeneration in the urochordate Botrylloides leachi. PLoS Biol 5:e71. https://doi.org/10.1371/journal.pbio.0050071
Rinkevich Y, Rinkevich B, Reshef R (2008) Cell signaling and transcription factor genes expressed during whole body regeneration in a colonial chordate. BMC Dev Biol 8:100. https://doi.org/10.1186/1471-213X-8-100
Rinkevich Y, Rosner A, Rabinowitz C et al (2010) Piwi positive cells that line the vasculature epithelium, underlie whole body regeneration in a basal chordate. Dev Biol 345:94–104. https://doi.org/10.1016/j.ydbio.2010.05.500
Rinkevich Y, Voskoboynik A, Rosner A et al (2013) Repeated, long-term cycling of putative stem cells between niches in a basal chordate. Dev Cell 24:76–88. https://doi.org/10.1016/j.devcel.2012.11.010
Roberts B, Davidson B, MacMaster G et al (2007) A complement response may activate metamorphosis in the ascidian Boltenia villosa. Dev Genes Evol 217:449–458. https://doi.org/10.1007/s00427-007-0157-0
Rosner A, Rinkevich B (2007) The DDX3 subfamily of the DEAD box helicases: divergent roles as unveiled by studying different organisms and in vitro assays. Curr Med Chem 14:2517–2525. https://doi.org/10.2174/092986707782023677
Rosner A, Rinkevich B (2011) VASA as a specific marker for germ cells lineage: in light of evolution. Trends Comp Biochem Physiol 15:1–15
Rosner A, Paz G, Rinkevich B (2006) Divergent roles of the DEAD-box protein BS-PL10, the urochordate homologue of human DDX3 and DDX3Y proteins, in colony astogeny and ontogeny. Dev Dyn 235:1508–1521. https://doi.org/10.1002/dvdy.20728
Rosner A, Moiseeva E, Rinkevich Y et al (2009) Vasa and the germ line lineage in a colonial urochordate. Dev Biol 331:113–128. https://doi.org/10.1016/j.ydbio.2009.04.025
Rosner A, Moiseeva E, Rabinowitz C, Rinkevich B (2013) Germ lineage properties in the urochordate Botryllus schlosseri - from markers to temporal niches. Dev Biol 384:356–374. https://doi.org/10.1016/j.ydbio.2013.10.002
Rosner A, Alfassi G, Moiseeva E et al (2014) The involvement of three signal transduction pathways in botryllid ascidian astogeny, as revealed by expression patterns of representative genes. Int J Dev Biol 58:669–676. https://doi.org/10.1387/ijdb.140114ar
Roy S, Gatien S (2008) Regeneration in axolotls: a model to aim for! Exp Gerontol 43:968–973. https://doi.org/10.1016/j.exger.2008.09.003
Rychel AL, Swalla BJ (2009) Regeneration in hemichordates and echinoderms. In: Stem cells in marine organisms. Springer Netherlands, Dordrecht, pp 245–265
Saito Y, Watanabe H (1985) Studies on Japanese compound styelid ascidians. IV. Three new species of the genus Botrylloides from the vicinity of Shimoda. Publ Seto Mar Biol Lab 30:227–240
Saito Y, Hirose E, Watanabe H (1994) Allorecognition in compound ascidians. Int J Dev Biol 38:237–247
Sánchez Alvarado A (2000) Regeneration in the metazoans: why does it happen? Bioessays 22:578–590
Sánchez Alvarado A, Tsonis PA (2006) Bridging the regeneration gap: genetic insights from diverse animal models. Nat Rev Genet 7:873–884. https://doi.org/10.1038/nrg1923
Satoh N (2003) The ascidian tadpole larva: comparative molecular development and genomics. Nat Rev Genet 4:285–295. https://doi.org/10.1038/nrg1042
Satoh N (2011) Tunicate embryos and cell specification. In: eLS. Wiley, Chichester
Savigny J-C (1816) Mémoires sur les animaux sans vertèbres. G. Dufour, Paris
Sawada H, Yokosawa H, Lambert CC (2001) The biology of ascidians. Springer, Tokyo
Seifert AW, Kiama SG, Seifert MG et al (2012) Skin shedding and tissue regeneration in African spiny mice (Acomys). Nature 489:561–565. https://doi.org/10.1038/nature11499
Slack JM (2017) Animal regeneration: ancestral character or evolutionary novelty? EMBO Rep 18:e201643795. https://doi.org/10.15252/embr.201643795
Stolfi A, Gandhi S, Salek F, Christiaen L (2014) Tissue-specific genome editing in Ciona embryos by CRISPR/Cas9. Development 141:4115–4120. https://doi.org/10.1242/dev.114488
Stoner DS, Rinkevich B, Weissman IL (1999) Heritable germ and somatic cell lineage competitions in chimeric colonial protochordates. Proc Natl Acad Sci USA 96:9148–9153. https://doi.org/10.1073/pnas.96.16.9148
Sunanaga T, Inubushi H, Kawamura K (2010) Piwi-expressing hemoblasts serve as germline stem cells during postembryonic germ cell specification in colonial ascidian, Botryllus primigenus. Develop Growth Differ 52:603–614. https://doi.org/10.1111/j.1440-169X.2010.01196.x
Tamilselvi M, Sivakumar V (2011) Distribution of alien tunicates (Ascidians) in Tuticorin coast, India. World J Zool 6:164–172
Tiozzo S, Brown FD, De Tomaso AW (2008a) Regeneration and stem cells in ascidians. In: Stem Cells. Springer Netherlands, Dordrecht, pp 95–112
Tiozzo S, Voskoboynik A, Brown FD, De Tomaso AW (2008b) A conserved role of the VEGF pathway in angiogenesis of an ectodermally-derived vasculature. Dev Biol 315:243–255. https://doi.org/10.1016/j.ydbio.2007.12.035
Tsonis PA (2000) Regeneration in vertebrates. Dev Biol 221:273–284. https://doi.org/10.1006/dbio.2000.9667
Voskoboynik A, Weissman IL (2015) Botryllus schlosseri, an emerging model for the study of aging, stem cells, and mechanisms of regeneration. Invertebr Reprod Dev 59:33–38. https://doi.org/10.1080/07924259.2014.944673
Voskoboynik A, Rinkevich B, Weiss A et al (2004) Macrophage involvement for successful degeneration of apoptotic organs in the colonial urochordate Botryllus schlosseri. J Exp Biol 207:2409–2416. https://doi.org/10.1242/jeb.01045
Voskoboynik A, Simon-Blecher N, Soen Y et al (2007) Striving for normality: whole body regeneration through a series of abnormal generations. FASEB J 21:1335–1344. https://doi.org/10.1096/fj.06-7337com
Voskoboynik A, Soen Y, Rinkevich Y et al (2008) Identification of the endostyle as a stem cell niche in a colonial chordate. Cell Stem Cell 3:456–464. https://doi.org/10.1016/j.stem.2008.07.023
Zaniolo G, Manni L, Burighel P (1994) Ovulation and embryo-parent relationships in Botrylloides leachi (Ascidiacea, Tunicata). Invertebr Reprod Dev 25:215–225. https://doi.org/10.1080/07924259.1994.9672388
Zondag LE, Rutherford K, Gemmell NJ, Wilson MJ (2016) Uncovering the pathways underlying whole body regeneration in a chordate model, Botrylloides leachi using de novo transcriptome analysis. BMC Genomics 17:114. https://doi.org/10.1186/s12864-016-2435-6
Acknowledgements
We would like to thank Aude Blanchoud for proofreading this review. SB was funded by the Swiss National Science Foundation (SNSF) grant number PZ00P3_17398. MJW was funded by a School of Biomedical Sciences Dean’s bequest grant. BR was funded by the Israel Academy of Sciences grant number 172/17. SB, MJW and BR were funded by a Royal Society of NZ Marsden fund grant (UOO1713).
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Blanchoud, S., Rinkevich, B., Wilson, M.J. (2018). Whole-Body Regeneration in the Colonial Tunicate Botrylloides leachii. In: Kloc, M., Kubiak, J. (eds) Marine Organisms as Model Systems in Biology and Medicine. Results and Problems in Cell Differentiation, vol 65. Springer, Cham. https://doi.org/10.1007/978-3-319-92486-1_16
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