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5 results on '"Deveci, Remziye"'

1. Morphological and ultrastructural characterization of sea urchin immune cells.

Author

Deveci R, Şener E, and İzzetoğlu S

Subjects
Animals, Microscopy, Electron, Transmission, Immune System cytology, Sea Urchins immunology
Abstract

The free circulating coelomocytes in the coelomic cavity of echinoderms are considered to be immune effectors by phagocytosis, encapsulation, cytotoxicity, and by the production of antimicrobial agents. Although echinoderms (especially sea urchin embryo) have been used as a model organisms in biology, no uniform criteria exist for classification of coelomocytes in echinoderms, and few studies have reported about the biological functions of their coelomocytes. Hence, we study the coelomocytes in the echinoid sea urchin, Paracentrotus lividus, and describe their morphological and ultrastructural features using light and transmission electron microscopes. We classify the coelomocytes of P. lividus into red spherule and colorless spherule cells, small cells, vibratile cells, and phagocytic cells; petaloid and filopodial cells. To our knowledge, this is the first report describing ultrastructural details of the coelomocytes of P. lividus., (© 2015 Wiley Periodicals, Inc.)

Published
2015
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2. Exploring the Candidate Terminal Glycan Profile in Neural Regeneration of the Sea Urchin Paracentrotus lividus, Using Lectin Blotting and Mass Spectrometry.

Author

Demir, Ramiz, Şahar, Umut, and Deveci, Remziye

Subjects
ELECTROSPRAY ionization mass spectrometry, GLYCANS, PARACENTROTUS lividus, SEA urchins, NERVOUS system regeneration, MASS spectrometry, CAPILLARY liquid chromatography
Abstract

Glycans are expressed as conjugates of glycoproteins, glycolipids, and proteoglycans. The huge diversity of glycans on glycoconjugates contributes to many biological processes, from glycan-based molecular recognition to developmental events, such as regeneration in the nervous system. Echinoderms, which have a close phylogenetic relationship with chordates, are an important group of marine invertebrates for body regeneration. Although many major roles of glycans on glycoconjugates are known, their role in the glycosylation profile of the nervous system in sea urchins is poorly understood. In this study, we aimed to determine the terminal glycan profile by lectin blotting and to quantify sialic acids by the capillary liquid chromatography electrospray ionization tandem mass spectrometry system in the nervous tissue of the sea urchin Paracentrotus lividus. We determined the N -acetyl-D-glucosamine, mannose, and sialic acids (mainly α2,3 linked) by lectin blotting and five types of sialic acids (N -glycolylneuraminic acid, N -acetylneuraminic acid, 9- O -acetyl- N -alycolylneuraminic acid, 5- N -acetyl-9- O -acetyl- N -acetylneuraminic acid, and di- O -acetylated- N -alycolylneuraminic acid) by capillary liquid chromatography electrospray ionization tandem mass spectrometry. This potential first description of the terminal glycan profile in the nervous system of the sea urchin is expected to help us understand its role in nervous system development and regeneration. [ABSTRACT FROM AUTHOR]

Published
2022
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3. Determining the monosaccharides of the sea urchin (Paracentrotus lividus) coelomocytes via the CapLC-ESI-MS/MS system and the lectin histochemistry.

Author

Şener, Ecem and Deveci, Remziye

Subjects
*MONOSACCHARIDES, *SEA urchins, *COELOMOCYTES, *HISTOCHEMISTRY, *LECTINS, *LIQUID chromatography-mass spectrometry
Abstract

The essential mechanism within immune systems is the recognition of pathogens and parasites by the immune system cells, which attach to their targets and destroy them. Glycans are fundamental macromolecular components of all cells, and are important in the vertebrate immunity. But, glycans have been investigated rarely in coelomocytes of echinoids. Hence, the aim of this study is to determine the monosaccharides which form glycan chains on the sea urchin immune system cells, coelomocytes, via analytical and lectin histochemistry methods. The study material is the coelomocytes obtained from adult sea urchin Paracentrotus lividus . In order to analyze the monosaccharides with the Capillary Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometry (CapLC-ESI-MS/MS) system, the samples underwent hydrolysation, reacetylation and derivatization steps. In order to determine the monosaccharides with the lectin histochemistry, the cells were incubated with fluorescein isothiocyanate (FITC) conjugated PNA, HPA, WGA-suc, WGA, and PSL lectins and then photographed with the fluorescence microscope. As a result of the CapLC-ESI-MS/MS analysis; mannose, ribose, N-acetylglucosamine, glucose, N-acetylgalactosamine, galactose, arabinose, xylose and fucose monosaccharides were detected. A peak area calculation analysis revealed the most prevalent saccharides as glucose, galactose and fucose, respectively. Lectin histochemistry came out with higher intensity emission signals obtained from the FITC-conjugated lectin WGA, which is specific to N-acetylglucosamine and sialic acid in comparison to the emission obtained from the sialic acid unspecific WGA-suc lectin. This finding indicates the existence of sialic acid within coelomocytes. Fluorescent emissions from other lectins were detected at lower levels. Determination of the monosaccharides which form glycan chains of the sea urchin coelomocytes and elucidating their similarities among other invertebrate and vertebrate systems is vital in terms of understanding the uncovered complex features of the immune systems of higher vertebrates. [ABSTRACT FROM AUTHOR]

Published
2015
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4. Determination of sialic acids in immune system cells (coelomocytes) of sea urchin, Paracentrotus lividus, using capillary LC-ESI-MS/MS.

Author

İzzetoğlu, Savaş, Şahar, Umut, Şener, Ecem, and Deveci, Remziye

Subjects
*SIALIC acids, *SEA urchins, *COELOMOCYTES, *PARACENTROTUS lividus, *IMMUNE response in fishes, *LIQUID chromatography-mass spectrometry, *ELECTROSPRAY ionization mass spectrometry
Abstract

Abstract: Coelomocytes are considered to be immune effectors of sea urchins. Coelomocytes are the freely circulating cells in the body fluid contained in echinoderm coelom and mediate the cellular defence responses to immune challenges by phagocytosis, encapsulation, cytotoxicity and the production of antimicrobial agents. Coelomocytes have the ability to recognize self from non-self. Considering that sialic acids play important roles in immunity, we determined the presence of sialic acid types in coelomocytes of Paracentrotus lividus. Homogenized coelomocytes were kept in 2 M aqueous acetic acid at 80 °C for 3 h to liberate sialic acids. Sialic acids were determined by derivatization with 1,2-diamino-4,5-methylenediaoxy-benzene dihydrochloride (DMB) followed by capillary liquid-chromatography-electrospray ionization/tandem mass spectrometry (CapLC-ESI-MS/MS). Standard sialic acids; Neu5Ac, Neu5Gc, KDN and bovine submaxillary mucin showing a variety of sialic acids were used to confirm sialic acids types. We found ten different types of sialic acids (Neu5Gc, Neu5Ac, Neu5Gc9Ac, Neu5Gc8Ac, Neu5,9Ac2, Neu5,7Ac2, Neu5,8Ac2, Neu5,7,9Ac3, Neu5Gc7,9Ac2, Neu5Gc7Ac) isolated in limited amounts from total coelomocyte population. Neu5Gc type of sialic acids in coelomocytes was the most abundant type sialic acid when compared with other types. This is the first report on the presence of sialic acid types in coelomocytes of P. lividus using CapLC-ESI-MS/MS-Ion Trap system (Capillary Liquid Chromatography-Electrospray Ionization/Tandem Mass Spectrometry). [Copyright &y& Elsevier]

Published
2014
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5. Denizkestanesi (Paracentrotus lividus)'un gamet yüzeylerinde sialik asit tiplerinin belirlenmesi ve döllenmedeki rolünün deneysel olarak araştırılması

Author

Yeşilyurt, Batuhan, Deveci, Remziye, Biyoloji Anabilim Dalı, and Fen Bilimleri Enstitüsü

Subjects
Deniz Bilimleri, Mass spectrometry, Spawning, FITC işaretli lektin, FITC Conjugated Lectin, Yumurta, Sialic acids, Sea Urchin Paracentrotus Lividus, Biochemistry, Sperm, CapLC-ESI-MS/MS, Sialic Acid, Biyokimya, Lectins, Egg, Marine Science, Denizkestanesi Paracentrotus lividus, Biology, Sea urchins, Sialik Asit, Biyoloji
Abstract

Hücresel tanıma olaylarında rol oynayan en önemli moleküllerin başında, bir uç şeker olan dokuz karbonlu modifiye şeker sialik asit (Sia) gelmektedir. Karbohidrat temelli döllenme mekanizması alanında bir çok çalışma yapılmasına karşın, Sia'nın döllenmedeki rolü konusunda hala aydınlanmamış kısımlar vardır. Bu nedenle çalışmanın amacı; farklı tiplerdeki sialik asitlerin döllenmedeki rolünün anlaşılması için, denizkestanesinin (Paracentrotus lividus) yumurta ve sperm yüzeylerinde ayrı ayrı belirlenecek sialik asit tiplerinden hangisi/hangilerinin döllenmede rol aldığının deneysel olarak gösterilmesidir. Çalışma materyali, denizkestanesi P.lividus'un ergin bireylerinden alınan yumurta ve sperm hücreleridir. Denizkestanelerinin vücut boşluğuna enjekte edilen KCl ile yumurta ve spermler elde edilmiştir. Yumurta yüzeyinde bulunan jel kılıf, asidik yöntem ile izole edilmiştir. Floresan mikroskobu ile gamet yüzeyindeki Sia'ları ve bağlanma özelliklerini belirlemek için FITC işaretli LPA, LFA, MAA, SNA, PSL, ve CCA lektinler kullanılmıştır. İzole jel kılıf örneklerindeki sialik asit tiplerinin Cap-LC-ESI-MS/MS sistemi ile analizi için örnekler hidroliz, reasetilasyon ve türevlendirme işlemlerinden sonra cihaza verilmiştir. Yumurta yüzeyindeki Sia'ların döllenmedeki önemini belirlemek için yumurtaların bulunduğu ortama tüm Sia'lara özgü LPA lektin çözeltisi verildikten sonra normal spermlerle bir araya getirilmiştir. Sperm yüzeyindeki Sia'ların döllenmedeki rolünü belirlemek için yumurtalar iki farklı tipte Sia (Neu5Ac ve Neu5Gc) ile yüzleştirildikten sonra işlem görmemiş normal spermlerle bir araya getirilmiştir. Sperm örneklerinde Sia belirlenmesi için uygulanan FITC işaretli lektinlerden en yoğun ışıma, Neu5Ac ve Neu5Gc Sia'larına özgü olan LFA ve Neu5Ac ye özgü olan LPA lektinlerinde gözlenmiştir. α2-3 bağına özgü MAA ve Neu5Ac α2-6 Gal, α1-4 GlcNAc bağ ve yapısına özgü PSL lektin ışımaları LFA ve LPA lektin ışımalarından daha zayıf, 9-O asetil-4-O asetil bağ ve yapılarına özgü CCA ve α2-6 bağına özgü SNA lektinlerin floresan ışımalarından daha yüksektir. İzole yumurta jel kılıfında Sia belirlenmesi için uygulanan FITC işaretli lektinlerden en yoğun ışıma, sperme benzer şekilde LFA ve LPA lektinlerinde gözlenmiştir. α2-6 bağına özgü SNA ve 9-O asetil-4-O asetil bağ ve yapılarına özgü CCA lektin ışımaları birbiriye hemen hemen aynı olup, LFA ve LPA lektin ışımalarından daha düşüktür. α2-3 bağına özgü MAA lektin ışıması tüm diğer lektin ışımalarından düşük olup, Neu5Ac α2-6 Gal, α1-4 GlcNAc bağ ve yapısına özgü PSL lektin ışıması gözlenmemiştir. İzole jel kılıfta CapLC-ESI-MS/MS sistemi ile analizi sonucu Neu5Gc, Neu5Ac, Neu5Gc7Ac, Neu5Gc8Ac, Neu5Gc9Ac, Neu5,7Ac2, Neu5,8Ac2, Neu5,9Ac2, Neu5Gc8,9Ac2, Neu5Gc7,9Ac2, Neu5AcS, Neu5GcS olmak üzere 12 tip Sia belirlenmiştir. Yumurta yüzeyindeki Sia'ların döllenmedeki rolünü araştırmak için yapılan deney sonucunda döllenme oranı %9,6 olarak hesaplanmıştır. Kontrol grubunda bu oran %99'dur. Sperm yüzeyindeki Sia'ların ve tiplerinin döllenmedeki rolü için yapılan deney sonucunda kontrol gruplarıyla deney grupları arasında anlamlı bir fark bulunamamıştır. Bu sonuç, Neu5Ac ve Neu5Gc'nin esansiyel olarak döllenme mekanizmasını kontrol etmediğini, diğer sialik asit çeşitlerinin de döllenmede spesifikliği sağlıyor olabileceğini işaret edebilir. Denizkestanesi gamet yüzeylerinde döllenmede rol oynayan sialik asit tiplerinin ve bağ özelliklerinin belirlenmesi, henüz tam olarak çözülmemiş karbohidrat temelli sperm yumurta ilişkisinin anlaşılması açısından önemlidir. Anahtar Kelimeler: Yumurta, Sperm, Sialik Asit, CapLC-ESI-MS/MS, FITC işaretli lektin, Denizkestanesi Paracentrotus lividus, Sialic acid is a terminal sugar of carbohydrate chains that participates in numerous biological events. Recent studies have explored the mechanism of carbohydrate-mediated fertilisation to understand the biochemistry of fertilisation. However, the type and quantity of sialic acid and the role of sialic acid during fertilisation remain unknown. Hence, the aim of this study is to determine the types and quantities of sialic acid in the gamete surface sea urchin, Paracentrotus lividus and investigation of in which type of them essential during fertilisation. The study materials are sperms and eggs obtained from adult sea urchin P. lividus. The gamets obtained via KCl injection to body cavity of sea urchins. We isolated egg jelly by using acidic method. In order to determine the Sia profile via the fluorescence method, the sperms and eggs are incubated with FITC conjugated LPA, LFA, MAA, SNA, PSL and CCA lectins. Then samples were photographed with a fluoresence microscope. In order to determine the types and quantities of Sia in the egg jelly coat, via using Cap-LC-ESI-MS/MS the samples underwnet hydrolyzation, reacetylation and derivatization steps followed by their flow throuh the device. In order to define the role of Sia on the egg jelly, we blocked the Sia of egg jelly coat with LPA lectins, and then we tested the egg's ability of fertilization with untreated sperms. In order to define the role of Sia on the surface of sperm, we treated eggs with two different Sia solutions (Neu5Ac and Neu5Gc) and tested their ability of fertilization with untreated sperms. Fluorescence analysis, which was performed in order to determine the Sia's on the sperm surface, came out with highest intensity emission signals obtained from FITC conjugated lectins LFA and LPA which specific to all types of Sia. The α2-3 bond specific MAA and Neu5Ac α2-6 Gal, α1-4 GlcNAc specific PSL lectins have a lower emision signals In comparison to the emission obtained from the LFA and LPA. The lowest emission signals obtained from, 9-O acetyl-4-O acetyl specific CCA and α2-6 bond specific SNA. in order to determine the Sia's on the egg jelly coat , came out with highest intensity emission signals obtained from FITC conjugated lectins LFA and LPA which specific to all types of Sia. The α2-6 bond specific SNA and 9-O acetyl-4-O acetyl specific CCA lectins have a lower emision signals In comparison to the emission obtained from the LFA and LPA. The lowest emission signals obtained from α2-3 bond specific MAA. There were no emission signals abtained Neu5Ac α2-6 Gal, α1-4 GlcNAc specific PSL. As a result of the Cap-LC-ESI-MS/MS analysis of egg jelly coat twelve different types of Sialic acid; Neu5Gc, Neu5Ac, Neu5Gc7Ac, Neu5Gc8Ac, Neu5Gc9Ac, Neu5,7Ac2, Neu5,8Ac2, Neu5,9Ac2, Neu5Gc8,9Ac2, Neu5Gc7,9Ac2, Neu5AcS, Neu5GcS were detected. As a result of the experiment which aimed to investigate the role of Sia on the egg jelly coat, the inhibiton with LPA lectins was successful about % 91,4. As a result of the experiment which aimed to investigate the role of Sia on the sperm surface, The inhibiton with two Sia solutions was not successful. The results highlights the importance of not only major types of Sia but also minor types of Sias might be ssential in fertilization process. Although echinoderm fertilisation has been studied extensively, our understanding of the mechanisms of carbohydrate-mediated fertilisation, and the role of sialic acid remains incomplete. The types and quantities of sialic acid that we detected in the egg jelly coat will aid in the discovery of new sialic acid-specific receptors on the sperm membrane. Keywords: Egg, Sperm, Sialic Acid, CapLC-ESI-MS/MS, FITC conjugated lectin, Sea urchin Paracentrotus lividus

Published
2015

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