Dynamic changes in sialic acid composition during fertilization and embryogenesis in the Sea Urchin Paracentrotus lividus (P. lividus)

Abstract

Purpose: The aim of this study was to investigate the dynamic changes in sialic acid composition during embryonic development of the sea urchin Paracentrotus lividus (P. lividus) and to evaluate stage-specific variations in sialic acid composition. Method: Sialic acid variants were analyzed in unfertilized eggs and embryos at the fertilized egg, 4-cell, 128-cell, blastula, gastrula, and pluteus stages using capillary LC–ESI–MS/MS following DMB derivatization. Relative abundances were determined from the peak areas of LC–ESI–MS/MS MRM chromatograms. Statistical differences among developmental stages were evaluated using one-way ANOVA followed by Tukey's multiple comparisons test. Findings: Seven sialic acid species, including Neu5Ac, Neu5Gc, and five O-acetylated derivatives, were identified throughout embryonic development. Neu5Gc was the predominant sialic acid in unfertilized eggs and exhibited a significant reduction immediately after fertilization (P < 0.0001), followed by a transient increase during early cleavage and blastula stages before declining markedly in gastrula and pluteus embryos. Neu5Ac remained relatively stable throughout early development but decreased significantly at the pluteus stage (P < 0.001). Several O-acetylated sialic acid derivatives displayed distinct stage-specific abundance patterns, indicating dynamic remodeling of sialic acid composition during embryogenesis. Conclusion: These findings demonstrate that sialic acid composition is developmentally regulated during P. lividus embryogenesis. The stage-specific alterations in Neu5Gc, Neu5Ac, and their O-acetylated derivatives suggest coordinated regulation of sialylation during early embryonic development and provide a foundation for future studies investigating the functional roles of sialic acid modifications in developmental processes.

Keywords

• sialic acids
• embryogenesis
• sea urchin
• glycobiology
• lc-ms/ms

Supporting Institution

The Scientific and Technological Research Council of Turkey (TÜBİTAK)

Project Number

2209-A

Ethical Statement

This study does not require ethical approval.

Thanks

The authors thank all contributors to this work.

Deniz Kestanesi Paracentrotus lividus (P. Lividus)'ta döllenme ve embriyogenez sırasında sialik asit kompozisyonundaki dinamik değişimler

Abstract

Amaç: Bu çalışmanın amacı, deniz kestanesi Paracentrotus lividus (P. lividus)'un embriyonik gelişimi sırasında sialik asit bileşimindeki dinamik değişimleri incelemek ve sialik asit bileşimindeki evreye özgü farklılıkları değerlendirmektir. Metod: DMB türevlendirmesini takiben kapiler LC–ESI–MS/MS kullanılarak; döllenmemiş yumurtalarda ve döllenmiş yumurta, 4-hücre, 128-hücre, blastula, gastrula ve pluteus evrelerindeki embriyolarda sialik asit varyantları analiz edilmiştir. Bağıl bolluklar, LC–ESI–MS/MS MRM kromatogramlarındaki pik alanlarından belirlenmiştir. Gelişim evreleri arasındaki istatistiksel farklılıklar, tek yönlü ANOVA ve ardından Tukey çoklu karşılaştırma testi kullanılarak değerlendirilmiştir. Bulgular: Embriyonik gelişim boyunca Neu5Ac, Neu5Gc ve beş O-asetillenmiş türev dahil olmak üzere yedi sialik asit türü tanımlanmıştır. Neu5Gc, döllenmemiş yumurtalarda baskın sialik asit türü olup döllenmeden hemen sonra önemli ölçüde azalmış (P < 0,0001); bunu erken bölünme ve blastula evrelerinde geçici bir artış izlemiş, ardından gastrula ve pluteus embriyolarında belirgin bir düşüş göstermiştir. Neu5Ac erken gelişim boyunca nispeten sabit kalmış ancak pluteus evresinde önemli ölçüde azalmıştır (P < 0,001). Çeşitli O-asetillenmiş sialik asit türevleri, embriyogenez sırasında sialik asit bileşiminin dinamik olarak yeniden düzenlendiğini gösteren, evreye özgü belirgin bolluk paternleri sergilemiştir. Sonuç: Bu bulgular, P. lividus embriyogenezi sırasında sialik asit bileşiminin gelişimsel olarak düzenlendiğini ortaya koymaktadır. Neu5Gc, Neu5Ac ve bunların O-asetillenmiş türevlerindeki evreye özgü değişimler, erken embriyonik gelişim sırasında sialilasyonun koordineli bir şekilde düzenlendiğini düşündürmekte ve gelişimsel süreçlerde sialik asit modifikasyonlarının işlevsel rollerini araştıracak gelecekteki çalışmalar için bir temel oluşturmaktadır.

Keywords

• sialik asitler
• embriyogenez
• deniz kestanesi
• glikobiyoloji
• lc-ms/ms

Supporting Institution

Türkiye Bilimsel ve Teknolojik Araştırma Kurumu (TÜBİTAK)

Project Number

2209-A

Ethical Statement

Çalışmanın yürütüldüğü dönemde, deniz kestanesi içeren deneyler için etik onay gerekli değildi.

Thanks

Yazarlar, bu çalışmaya katkıda bulunan herkese teşekkür ederler.

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