Practical Guide for Ascidian Microinjection: Phallusia mammillata
| Autor: | Alex McDougall, Hitoyoshi Yasuo |
|---|---|
| Přispěvatelé: | Laboratoire de Biologie du Développement de Villefranche sur mer (LBDV), Observatoire océanologique de Villefranche-sur-mer (OOVM), Centre National de la Recherche Scientifique (CNRS)-Institut national des sciences de l'Univers (INSU - CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Institut national des sciences de l'Univers (INSU - CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Institut de la Mer de Villefranche (IMEV), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS) |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Phallusia biology Phallusia mammillata Optical transparency Functional genes biology.organism_classification Cell biology 03 medical and health sciences [SDV.BDD.EO]Life Sciences [q-bio]/Development Biology/Embryology and Organogenesis 030104 developmental biology Live cell imaging embryonic structures Fluorescent protein Microinjection ComputingMilieux_MISCELLANEOUS |
| Zdroj: | Transgenic Ascidians ISBN: 9789811075445 Adv Exp Med Biol Adv Exp Med Biol, pp.15-24, 2018, ⟨10.1007/978-981-10-7545-2_3⟩ |
| Popis: | Phallusia mammillata has recently emerged as a new ascidian model. Its unique characteristics, including the optical transparency of eggs and embryos and efficient translation of exogenously introduced mRNA in eggs, make the Phallusia system suitable for fluorescent protein (FP)-based imaging approaches. In addition, genomic and transcriptomic resources are readily available for this ascidian species, facilitating functional gene studies. Microinjection is probably the most versatile technique for introducing exogenous molecules such as plasmids, mRNAs, and proteins into ascidian eggs/embryos. However, it is not practiced widely within the community; presumably, because the system is rather laborious to set up and it requires practice. Here, we describe in as much detail as possible two microinjection methods that we use daily in the laboratory: one based on an inverted microscope and the other on a stereomicroscope. Along the stepwise description of system setup and injection procedure, we provide practical tips in the hope that this chapter might be a useful guide for introducing or improving a microinjection setup. |
| Databáze: | OpenAIRE |
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