The role of metalloproteases in fertilisation in the ascidian Ciona robusta

In the ascidian Ciona robusta (formerly C. intestinalis type A), the mechanism underlying sperm penetration through the egg investment remains unknown. We previously reported that proteins containing both an astacin metalloprotease domain and thrombospondin type 1 repeats are abundant in the sperm s...

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Bibliographic Details
Published in:Scientific reports 2019-01, Vol.9 (1), p.1009-1009, Article 1009
Main Authors: Nakazawa, Shiori, Shirae-Kurabayashi, Maki, Sawada, Hitoshi
Format: Article
Language:English
Subjects:
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Animals
Astacin
Ciona intestinalis
Ciona robusta
Developmental stages
Egg Proteins - metabolism
Eggs
Female
Fertilization
Humanities and Social Sciences
Larval development
Male
Metalloproteases - physiology
Metalloproteinase
multidisciplinary
Proteins
Science
Science (multidisciplinary)
Sperm
Sperm-Ovum Interactions
Spermatozoa - metabolism
Thrombospondin
Vitelline Membrane - metabolism
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Summary:In the ascidian Ciona robusta (formerly C. intestinalis type A), the mechanism underlying sperm penetration through the egg investment remains unknown. We previously reported that proteins containing both an astacin metalloprotease domain and thrombospondin type 1 repeats are abundant in the sperm surface protein-enriched fraction of C. robusta . Here we investigated the involvement of those proteins in fertilisation. We refined the sequences of astacin metalloproteases, confirmed that five of them are present in the sperm, and labelled them as t unicate as tacin and t hrombospondin type 1 repeat-containing (Tast) proteins. Fertilisation of C. robusta eggs was potently inhibited by a metalloprotease inhibitor GM6001. The eggs cleaved normally when they were vitelline coat-free or the inhibitor was added after insemination. Furthermore, vitelline coat proteins were degraded after incubation with intact sperm. These results suggest that sperm metalloproteases are indispensable for fertilisation, probably owing to direct or indirect mediation of vitelline-coat digestion during sperm penetration. TALEN-mediated knockout of Tast genes and the presence of GM6001 impaired larval development at the metamorphic stage, suggesting that Tast gene products play a key role in late development.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-018-37721-1