Comparative Proteomic Analysis of Buffalo Oocytes Matured in vitro Using iTRAQ Technique

Abstract

To investigate the protein profiling of buffalo oocytes at the germinal vesicle (GV) stage and metaphase II (MII) stage, an iTRAQ-based strategy was applied. A total of 3,763 proteins were identified, which representing the largest buffalo oocytes proteome dataset to date. Among these proteins identified, 173 proteins were differentially expressed in GV oocytes and competent MII oocytes, and 146 proteins were differentially abundant in competent and incompetent matured oocytes. Functional and KEGG pathway analysis revealed that the up-regulated proteins in competent MII oocytes were related to chromosome segregation, microtubule-based process, protein transport, oxidation reduction, ribosome, and oxidative phosphorylation, etc., in comparison with GV and incompetent MII oocytes. This is the first proteomic report on buffalo oocytes from different maturation stages and developmental competent status. These data will provide valuable information for understanding the molecular mechanism underlying buffalo oocyte maturation, and these proteins may potentially act as markers to predict developmental competence of buffalo oocyte during in vitro maturation.

Compared proteomic analysis of 8‐ and 32‐week‐old postnatal porcine ovaries

“…Li et al explored the expression profiles of mRNAs, lncRNAs, and circRNAs in male and female mouse germline stem cells by high‐throughput sequencing . Regarding proteomics of female germ cells, Chen et al analysed buffalo oocytes matured in vitro by using the iTRAQ technique . Virant‐Klun et al identified maturation‐specific proteins in human oocytes by single‐cell proteomics .…”

Maturação oocitária: Eventos que suportam etapas subsequentes do desenvolvimento

“…Competence development of oocytes is primarily dependent on the biochemical and structural transformations that occur in the cytoplasm (Ferreira et al, 2009;Khan et al, 2016). These transformations, involved in maturation, are highly intricate processes involving several simultaneous events, such as protein syntheses, molecular modifications, migration and reorganization of organelles in the cytoplasm (Chen et al, 2016;Khan et al, 2016). These changes in the oocyte cytoplasm will be discussed in more detail in the following sections.…”

“…However, gene transcription ceases before ovulation, so the oocyte, zygote, and embryo (Figure 3), with less than 16 blastomeres (bovine), are dependent on the pool of mRNAs and proteins accumulated during the period of growth and maturation (Meirelles et al, 2004;Salilew-Wondim et al, 2018). Oocytes depend on the synthesis and activation of a variety of proteins so they can enter and continue the process of oocyte maturation (Soeda et al, 2013;Chen et al, 2016).…”

Tissue‐based quantitative proteomics to screen and identify the potential biomarkers for early recurrence/metastasis of esophageal squamous cell carcinoma

“…The iTRAQ method is a highly sensitive proteomic platform, which is characterized by high proteome coverage and labeling efficiency, and has no side effects on analytical or biochemical properties of the labeled proteins or peptides [11,12]. However, the iTRAQ technique is proved to be more suitable for initial biomarkers discovery, and the results would require validation through a large-scale screening [13,14]. In our study, we presented details of proteome variation in the different ESCC tumor tissues based on an iTRAQ with reversedphase liquid chromatography-mass spectrometry (RPLC-MS) approach, and further validated the observations by tissue microarray (TMA) based on large samples immunohistochemistry.…”