Abstract
Proper maturation of the mammalian oocyte is a compound processes determining successful monospermic fertilization, however the number of fully mature porcine oocytes is still unsatisfactory. Since oocytes’ maturation and fertilization involve cellular adhesion and membranous contact, the aim was to investigate cell adhesion ontology group in porcine oocytes. The oocytes were collected from ovaries of 45 pubertal crossbred Landrace gilts and subjected to two BCB tests. After the first test, only granulosa cell-free BCB+ oocytes were directly exposed to microarray assays and RT-qPCR (“before IVM” group), or first in vitro matured and then if classified as BCB+ passed to molecular analyses (“after IVM” group). As a result, we have discovered substantial down-regulation of genes involved in adhesion processes, such as: organization of actin cytoskeleton, migration, proliferation, differentiation, apoptosis, survival or angiogenesis in porcine oocytes after IVM, compared to oocytes analyzed before IVM. In conclusion, we found that biological adhesion may be recognized as the process involved in porcine oocytes’ successful IVM. Down-regulation of genes included in this ontology group in immature oocytes after IVM points to their unique function in oocyte’s achievement of fully mature stages. Thus, results indicated new molecular markers involved in porcine oocyte IVM, displaying essential roles in biological adhesion processes.
Highlights
During mammalian folliculogenesis and oogenesis the “ovarian follicle” differentiates from primordial, primary and secondary, into the fully organized Graafian follicle. These unique stages are accompanied by substantial growth and development of follicle [1], which is composed of differentiating theca cells (TCs) and granulosa cells (GCs)
The oocyte maturation is divided into two crucial steps: nuclear, which is associated with achievement of proper chromosomal configuration by the oocyte, and cytoplasmic, which is accompanied by storage of RNA and proteins for further embryo growth [3,4]
DAVID (Database for Annotation, Visualization and Integrated Discovery) software was used for extraction of the genes belonging to “Biological adhesion” gene ontology Biological Process term (GO BP)
Summary
During mammalian folliculogenesis and oogenesis the “ovarian follicle” differentiates from primordial, primary and secondary, into the fully organized Graafian follicle These unique stages are accompanied by substantial growth and development of follicle [1], which is composed of differentiating theca cells (TCs) and granulosa cells (GCs). The oocyte maturation is divided into two crucial steps: nuclear, which is associated with achievement of proper chromosomal configuration by the oocyte, and cytoplasmic, which is accompanied by storage of RNA and proteins for further embryo growth [3,4] It was well recognized in several species of mammals that only properly structuralized mature oocyte (MII) may be successfully fertilized by spermatozoon, and each stage of early folliculogenesis and oogenesis influences further embryo growth and development [5,6,7]. Our previous experiments indicated an increased expression of genes involved in “Bone development”, “Cellular components of morphogenesis”, “BMP signaling pathway”, and “Cell migration”, which were intensively regulated during in vitro maturation of porcine oocytes (IVM) [8,9,10,11,12]
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