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Comparative proteomic analysis of eggs and ovarian fluid in Siberian sturgeon: Insights into composition and functional significance

ABSTRACT: Sturgeon species, considered living fossils, exhibit unique reproductive characteristics, making it crucial to comprehend the molecular processes governing the formation and quality of their eggs. However, there is a notable lack of comprehensive data concerning the protein composition analysis of sturgeon eggs and ovarian fluid (OF) and their functional significance. To address this knowledge gap, this study aimed to conduct a comprehensive comparative proteomic analysis of Siberian sturgeon eggs and OF using liquid chromatography-mass spectrometry (LC-MS/MS). A total of 566 proteins were identified in eggs, while 617 proteins were identified in OF, with 772 proteins showing differential abundance. Among the differentially abundant proteins, 407 were more abundant in eggs, while 365 showed higher abundance in OF. Furthermore, we identified 288 proteins specific to eggs and 339 proteins specific to OF, along with the top ten most abundant proteins in each. Functional annotation analysis unveiled enriched metabolic pathways, such as oxidative phosphorylation and fatty acid metabolism, as well as protein ubiquitination and translation, in eggs. Conversely, ovarian fluid proteins primarily associated with immune system processes, including the complement and coagulation cascade, neutrophil and leukocyte-mediated immunity, cholesterol metabolism, and regulation of actin cytoskeleton. This study presents the first comprehensive characterization of the protein composition of sturgeon ovarian fluid and eggs, shedding light on their distinct functional roles. The findings not only advance our understanding of sturgeon reproduction but also shed light on egg-OF signaling and origin of the OF proteins. Moreover the identified proteins offer potential biomarkers for predicting egg quality contributing to the development of effective breeding strategies for sturgeon species.

INSTRUMENT(S): Orbitrap Exploris 480

ORGANISM(S): Acipenser Baerii

TISSUE(S): Roe, Ovarian Fluid

SUBMITTER: Agata Malinowska

LAB HEAD: Mariola A. Dietrich

PROVIDER: PXD044168 | Pride | 2023-12-18

REPOSITORIES: Pride

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Project description:The biology and physiology of Siberian sturgeon reproduction differ substantially from teleost fish. Seminal plasma support and protect the viability, motility and fertilizing capacity of spermatozoa by creating the optimal environment for the storage. The present study for the first time described in-depth proteomic characterization of Siberian sturgeon seminal plasma using gel based and gel-free proteomic approaches: LC-MS/MS, 2DE and 2D blue native (BN)/SDS-PAGE. LC-MS/MS allowed to identify 657 proteins, which were classified according to their functions and pathways. 2DE visualized 339 spots corresponding to 76 unique proteins (almost 60% were present in various proteoforms). For the first time we demonstrated interactions between seminal proteins; four (C1-C4) multiprotein complexes were identified after 2D BN/SDS-PAGE; composed of (C1) serotransferrin-2 (TF) and fish-egg lectin (FEL); (C2) serum albumin 2 (ALB) and retinol-binding protein 4 (RBP4); (C3) ALB and TF (C3) and (C4) apolipoprotein A-I (APOA1), riboflavin-binding protein (RfBP) and myoglobin (MB). BN-SDS-PAGE also revealed that immunoglobulin and TF formed homomultimeric (dimer, trimer, tetramer and pentamer) complexes in seminal plasma. ALB, TF, Beta-Ala-His dipeptidase, glyceraldehyde-3-phosphate dehydrogenase, IGH, APOA1, hemopexin, type-4 ice-structuring protein, FEL, RfBP and glycogen phosphorylase, liver form-like were selected as major seminal plasma proteins. The functional analysis of identified proteins indicated their involvement mainly in immune system process and response to stimulus, metabolism, vesicle-mediated transport, proteolysis, and catherin binding involved in cell-cell adhesion. Moreover, 67 proteins were associated with reproductive processes, including spermatogenesis, fertilization, acrosomal reaction and sperm motility. Comparative proteomic analysis of sturgeon seminal plasma showed common proteins with fish and human and 150 proteins specific for sturgeon which may reflect the specificity of sturgeon reproduction. To conclude, this integrative proteomic view lead to deeper insight into physiological function of seminal plasma and processes occurred in sturgeon reproductive tract.

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Comparative proteomic analysis of eggs and ovarian fluid in Siberian sturgeon: Insights into composition and functional significance

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