Abstract
Ascidians are basal chordates that have become increasingly important for understanding chordate evolution. They comprise three orders. In the orders Phlebobranchia and Stolidobranchia, most species freely spawn eggs and sperm, whereas members of the order Aplousobranchia form colonies that brood their eggs and broadcast sperm. In the two free spawning orders, eggs and sperm are easily obtained for in vitro fertilizations. In the third order, slices of colonies yield gametes and embryos of all stages. Methods are described for obtaining gametes, performing fertilizations, and culturing embryos. Also included are methods for removing follicle cells and vitelline coats from oocytes.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Delsuc F et al (2006) Tunicates and not cephalochordates are the closest living relatives of vertebrates. Nature 439:965–968
Lambert CC (2009) Ascidian follicle cells: multifunctional adjuncts to maturation and development. Dev Growth Differ 51:677–686
Lahille F (1886) Sur le classification des Tuniciers. CR Acad Sci Paris CII:1573–1575
Bishop JD, Pemberton AJ (2006) The third way: spermcast mating in sessile marine invertebrates. Integr Comp Biol 46:398–406
Saito Y, Mukai H, Watanabe H (1981) Studies on Japanese compound styelid ascidians II. A new species of the genus Botrylloides and redescription of B. violaceus Oka. Publ Seto Mar Biol Lab 26:357–368
Scott SF (1945) The developmental history of Amaroecium constellatum 1. Early embryonic development. Biol Bull 88:126–138
Scott SF (1946) The developmental history of Amaroecium constellatum. II. Organogenesis of the larval action system. Biol Bull 91:66–80
Watanabe H, Lambert CC (1973) Larva release in response to light by the compound ascidians Distaplia occidentalis and Metandrocarpa taylori. Biol Bull 144: 556–566
Lambert CC, Brandt CL (1967) The effect of light on the spawning of Ciona intestinalis. Biol Bull 132:222–228
Lambert G, Lambert CC, Abbott DP (1981) Corella species in the American Pacific Northwest: distinction of C. inflata Huntsman, 1912 from C. willmeriana Herdman, 1898 (Ascidiacea, Phlebobranchia). Can J Zool 59: 1493–1504
Whittingham DG (1967) Light-induction of shedding of gametes in Ciona intestinalis and Molgula manhattensis. Biol Bull 132:292–298
Byrd J, Lambert CC (2000) Mechanism of the block to hybridization and selfing between the sympatric ascidians Ciona intestinalis and Ciona savignyi. Mol Reprod Dev 55:109–116
West AB, Lambert CC (1976) Control of spawning in the tunicate Styela plicata by variations in a natural light regime. J Exp Zool 195:263–270
Lambert CC (2005) Signaling pathways in ascidian oocyte maturation: effects of various inhibitors and activators on germinal vesicle breakdown. Dev Growth Differ 47:265–272
McDougall A, Sardet C, Lambert CC (1995) Different calcium-dependent pathways control fertilisation-triggered glycoside release and the cortical contraction in ascidian eggs. Zygote 3:251–258
Lambert CC et al (1997) Ascidian eggs block polyspermy by two independent mechanisms: one at the plasma membrane, the other involving the follicle cells. Mol Reprod Dev 48:137–143
Fuke M (1983) Self and non-self recognition between gametes of the ascidian, Halocynthia roretzi. Roux’s Arch Dev Biol 192:347–352
Lambert CC (2008) Signaling pathways in ascidian oocyte maturation: the role of cyclic AMP and follicle cells in germinal vesicle breakdown. Dev Growth Differ 50:181–188
Cuomo A et al (2006) Ca2+ and Na+ current patterns during oocyte maturation, fertilization, and early developmental stages of Ciona intestinalis. Mol Reprod Dev 73:501–511
Sardet C et al (2011) Embryological methods in ascidians: the Villefranche protocols. Methods Mol Biol 770:365–400
Mita-Miyazawa I, Ikegami S, Satoh N (1985) Histospecific acetylcholinesterase development in the presumptive muscle cells isolated from 16-cell-stage ascidian embryos with respect to the number of DNA replications. J Embryol Exp Morphol 87:1–12
Zalocar M, Sardet C (1984) Tracing of cell lineage in embryonic development of Phallusia mammillata (Ascidia) by vital staining of mitochondria. Dev Biol 102:195–205
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media, New York
About this protocol
Cite this protocol
Lambert, C.C. (2014). Obtaining Gametes and Embryos of Ascidians. In: Carroll, D., Stricker, S. (eds) Developmental Biology of the Sea Urchin and Other Marine Invertebrates. Methods in Molecular Biology, vol 1128. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-974-1_2
Download citation
DOI: https://doi.org/10.1007/978-1-62703-974-1_2
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-973-4
Online ISBN: 978-1-62703-974-1
eBook Packages: Springer Protocols