"Cell Migration" Is the Ontology Group Differentially Expressed in Porcine Oocytes Before and After In Vitro Maturation: A Microarray Approach.

Maturation of cumulus-oocyte complexes (COCs) is crucial for further successful monospermic fertilization, embryo growth, and implantation. All these events are accompanied by proliferation and differentiation of cumulus cells. The migration of COCs to the oviduct after ovulation and the interaction between female gametes and/or embryos with maternal tissues are still poorly recognized on the molecular level. This study was aimed to first demonstrate the mRNA expression profile of cell migration markers during different stages of porcine oocytes maturation and developmental capability in vitro. The COCs were collected from a total of 45 pubertal crossbred Landrace gilts, brilliant cresyl blue (BCB) stained, and analyzed before (n = 150) or after (n = 150) in vitro maturation (IVM). Using the Affymetrix® Porcine Gene 1.1 ST Array, the expression profile of 12,258 porcine transcripts was examined. We found nine genes involved in cell migration mechanisms, that is, PLD1, KIT, LAMA2, MAP3K1, VEGFA, TGFBR3, INSR, TPM1, and RTN4. These genes were upregulated in porcine oocytes before IVM as compared with post-IVM expression analysis. Moreover, important mechanisms of biological interaction between VEGFA-KIT and VEGFA-INSR were also observed. The upregulation and/or downregulation of selected mRNAs expression after microarray assays was checked and approved by real-time quantitative polymerase chain reaction. We suggest that several genes, including LAMA2 or TPM1, encode proteins participating in the formation of the oocyte's protein architecture such as microtubules and kinetochore reorganization. As the expression of all "migration regulatory genes" investigated in this study was significantly upregulated in oocytes before IVM, we conclude that they may contribute to the maturational capability of porcine oocytes. However, migration potency of COCs is not accompanied by achievement of the MII stage by porcine oocytes in vitro. The investigated genes such as PLD1, KIT, LAMA2, MAP3K1, VEGFA, TGFBR3, INSR, TPM1, and RTN4 may be recognized as a new marker of porcine oocytes maturational competence during in vitro culture.

[1]  H. Piotrowska,et al.  The role of TGF superfamily gene expression in the regulation of folliculogenesis and oogenesis in mammals: a review , 2018 .

[2]  A. Baccarelli,et al.  Extracellular vesicles: roles in gamete maturation, fertilization and embryo implantation. , 2015, Human reproduction update.

[3]  H. Valadi,et al.  Molecular characterization of exosomes and their microRNA cargo in human follicular fluid: bioinformatic analysis reveals that exosomal microRNAs control pathways involved in follicular maturation. , 2014, Fertility and sterility.

[4]  J. Ross,et al.  Cytokines from the pig conceptus: roles in conceptus development in pigs , 2014, Journal of Animal Science and Biotechnology.

[5]  H. Piotrowska,et al.  Study on connexin gene and protein expression and cellular distribution in relation to real-time proliferation of porcine granulosa cells. , 2014, Journal of biological regulators and homeostatic agents.

[6]  S. Namgoong,et al.  Effects of growth differentiation factor 9 and bone morphogenetic protein 15 on the in vitro maturation of porcine oocytes. , 2014, Reproduction in domestic animals = Zuchthygiene.

[7]  Thomas E. Spencer,et al.  Exosomal and Non-Exosomal Transport of Extra-Cellular microRNAs in Follicular Fluid: Implications for Bovine Oocyte Developmental Competence , 2013, PloS one.

[8]  H. Piotrowska,et al.  Real-time proliferation of porcine cumulus cells is related to the protein levels and cellular distribution of Cdk4 and Cx43. , 2013, Theriogenology.

[9]  H. Piotrowska,et al.  Short-term Cultivation of Porcine Cumulus Cells Influences the Cyclin-dependent Kinase 4 (Cdk4) and Connexin 43 (Cx43) Protein Expression—A Real-time Cell Proliferation Approach , 2013, The Journal of reproduction and development.

[10]  A. Uyar,et al.  Cumulus and granulosa cell markers of oocyte and embryo quality. , 2013, Fertility and sterility.

[11]  E. De Ponti,et al.  Cumulus cell-oocyte complexes retrieved from antral follicles in IVM cycles: relationship between COCs morphology, gonadotropin priming and clinical outcome , 2012, Journal of Assisted Reproduction and Genetics.

[12]  G. Bouma,et al.  Cell-Secreted Vesicles in Equine Ovarian Follicular Fluid Contain miRNAs and Proteins: A Possible New Form of Cell Communication Within the Ovarian Follicle1 , 2012, Biology of reproduction.

[13]  Y. Soong,et al.  Regulation of Oocyte and Cumulus Cell Interactions by Intermedin/Adrenomedullin 2* , 2011, The Journal of Biological Chemistry.

[14]  A. Liekens,et al.  BioGraph: unsupervised biomedical knowledge discovery via automated hypothesis generation , 2011, Genome Biology.

[15]  Nicola Bernabò,et al.  The spermatozoa caught in the net: the biological networks to study the male gametes post-ejaculatory life , 2010, BMC Systems Biology.

[16]  B. Vanderhyden,et al.  Bidirectional communication between oocytes and follicle cells: ensuring oocyte developmental competence. , 2010, Canadian journal of physiology and pharmacology.

[17]  A. Wehrend,et al.  Ciliary Transport, Gamete Interaction, and Effects of the Early Embryo in the Oviduct: Ex Vivo Analyses Using a New Digital Videomicroscopic System in the Cow1 , 2009, Biology of reproduction.

[18]  David Bryant,et al.  DAVID Bioinformatics Resources: expanded annotation database and novel algorithms to better extract biology from large gene lists , 2007, Nucleic Acids Res..

[19]  A. J. Watson Oocyte cytoplasmic maturation: a key mediator of oocyte and embryo developmental competence. , 2007, Journal of animal science.

[20]  A. Spradling,et al.  Mouse oocytes within germ cell cysts and primordial follicles contain a Balbiani body , 2007, Proceedings of the National Academy of Sciences.

[21]  K. Wigglesworth,et al.  The preantral granulosa cell to cumulus cell transition in the mouse ovary: development of competence to undergo expansion. , 2006, Developmental biology.

[22]  Christian von Mering,et al.  STRING: known and predicted protein–protein associations, integrated and transferred across organisms , 2004, Nucleic Acids Res..

[23]  J. Denegre,et al.  Oocyte-dependent activation of mitogen-activated protein kinase (ERK1/2) in cumulus cells is required for the maturation of the mouse oocyte-cumulus cell complex. , 2003, Developmental biology.

[24]  H. Croxatto,et al.  Physiology of gamete and embryo transport through the fallopian tube. , 2002, Reproductive biomedicine online.

[25]  S. Hammond,et al.  Human ADP-ribosylation Factor-activated Phosphatidylcholine-specific Phospholipase D Defines a New and Highly Conserved Gene Family (*) , 1995, The Journal of Biological Chemistry.

[26]  F. Le Gal,et al.  Protein phosphorylation patterns during in vitro maturation of the goat oocyte , 1993, Molecular reproduction and development.

[27]  S. Dieleman,et al.  Morphology of preovulatory bovine follicles as related to oocyte maturation. , 1991, Theriogenology.