Abstract
In the last decades, the cephalochordate amphioxus has reached a peculiar place in research laboratories as an excellent animal model to answer Evo/Devo questions. Nevertheless, mainly due to its restricted spawning season and to the small size of its embryos, only a few basic techniques in developmental biology could be used until recently. Fortunately, these last years, and thanks to the development of high-throughput techniques, new technical approaches have been possible, such as comparative transcriptomics and/or genomics. However, classic micromanipulation techniques are still difficult to apply. Here we present simple protocols for the manipulation of amphioxus embryos. First, we present the spawning induction method used with the European amphioxus species Branchiostoma lanceolatum. Second, we explain simple methods to manipulate the developing amphioxus embryo during the first steps of its development (before the hatching stage). These methods open many technical possibilities for future functional studies. Thus, we present here a simple technique to efficiently dechorionate a large number of embryos, we detail a protocol for the dissociation of cells during the first steps of the embryonic development and, finally, we describe micromanipulation approaches for tissue isolation during the gastrula stage.
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References
Bourlat SJ, Juliusdottir T, Lowe CJ, Freeman R, Aronowicz J, Kirschner M et al (2006) Deuterostome phylogeny reveals monophyletic chordates and the new phylum Xenoturbellida. Nature 444(7115):85–88
Delsuc F, Brinkmann H, Chourrout D, Philippe H (2006) Tunicates and not cephalochordates are the closest living relatives of vertebrates. Nature 439(7079):965–968
Bertrand S, Escriva H (2011) Evolutionary crossroads in developmental biology: amphioxus. Development 138(22):4819–4830
Poss SG, Boschung HT (1996) Lancelets (Cephalochordata: Branchiostomatidae): how many species are valid? Isr J Zool 42(sup1):S13–S66
Fuentes M, Benito E, Bertrand S, Paris M, Mignardot A, Godoy L et al (2007) Insights into spawning behavior and development of the European amphioxus (Branchiostoma lanceolatum). J Exp Zool B Mol Dev Evol 308B(4):484–493
Le Petillon Y, Luxardi G, Scerbo P, Cibois M, Leon A, Subirana L et al (2017) Nodal–activin pathway is a conserved neural induction signal in chordates. Nat Ecol Evol 1(8):1192–1200
Holland LZ, Yu J-K (2004) Cephalochordate (amphioxus) embryos: procurement, culture, and basic methods. Methods Cell Biol 74:195–215
Tung TC, Wu SC, Tung YY (1960) The developmental potencies of the blastomere layers in Amphioxus egg at the 32-cell stage. Sci Sin 9:119–141
Tung TC, Wu SC, Tung YF (1958) The development of isolated blastomeres of Amphioxus. Sci Sin 7(12):1280–1320
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Le Petillon, Y., Bertrand, S., Escrivà, H. (2020). Spawning Induction and Embryo Micromanipulation Protocols in the Amphioxus Branchiostoma lanceolatum. In: Sprecher, S. (eds) Brain Development. Methods in Molecular Biology, vol 2047. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9732-9_19
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DOI: https://doi.org/10.1007/978-1-4939-9732-9_19
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-9731-2
Online ISBN: 978-1-4939-9732-9
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