Restricted accessResearch articleFirst published online 2019-06
Genes Involved in the Processes of Cell Proliferation,Migration,Adhesion,and Tissue Development as New Potential Markers of Porcine Granulosa Cellular Processes In Vitro : A Microarray Approach
Proper course of folliculogenesis and oogenesis have an enormous impact on female fertility. Both processes take place in the ovary and involve not only the maturing germ cell, but also few types of somatic cells that assist the ovarian processes and mediate the dialog with the oocyte. These cells, granulosa and theca, are heavily involved in essential reproductive processes, such as ovulation, fertilization, and embryo implantation. In this study, we have used the expressive microarray approach to analyze the transcriptome of porcine granulosa cells, during short-term in vitro culture. We have further selected differentially expressed gene ontologies, involved in cell proliferation, migration, adhesion, and tissue development, namely, “cell–cell adhesion,” “cell motility,” “cell proliferation,” “tissue development,” and “tissue migration” to screen them for the possibility of discovery of new markers of those processes. A total of 303 genes, expression of which varied significantly in different culture periods and belonged to the analyzed ontology groups, were detected, of which 15 that varied the most (between 0 and 48 h of culture) were selected for validation. As the validation confirmed the transcriptomic patterns, 10 genes of biggest changes in expression (CAV1, IGFBP5, ITGB3, FN1, ITGA2, LAMB1, POSTN, FAM83D, KIF14, and CDK1) were analyzed, described, and referred to the context of the study, with the most promising new markers and further proof for the viability of the currently recognized ones detailed. Overall, the study provided valuable insight into the molecular functioning of in vitro granulosa cell cultures.
Get full access to this article
View all access options for this article.
References
1.
AdashiE.Y., ResnickC.E., PayneD.W., RosenfeldR.G., MatsumotoT., HunterM.K., et al. (1997). The mouse intraovarian insulin-like growth factor I system: departures from the rat paradigm. Endocrinology, 138, 3881–3890.
2.
AdhikariD., BusayavalasaK., ZhangJ., HuM., RisalS., BayazitM.B., et al. (2016). Inhibitory phosphorylation of Cdk1 mediates prolonged prophase I arrest in female germ cells and is essential for female reproductive lifespan. Cell Res, 26, 1212–1225.
3.
AsemE.K., CarnegieJ.A., and TsangB.K. (1992). Fibronectin production by chicken granulosa cells in vitro: effect of follicular development. Acta Endocrinol, 127, 466–470.
4.
BajtM.L., GinsbergM.H., FrelingerA.L., 3rd, BerndtM.C., and LoftusJ.C. (1992). A spontaneous mutation of integrin alpha IIb beta 3 (platelet glycoprotein IIb-IIIa) helps define a ligand binding site. J Biol Chem, 267, 3789–3794.
5.
BryjaA., Dyszkiewicz-KonwińskaM., JankowskiM., CelichowskiP., StefańskaK., Chamier-GliszczyńskaA., et al. (2018). Ion homeostasis and transport are regulated by genes differentially expressed in porcine buccal pouch mucosal cells during long-term culture in vitro—a microarray approach. Med J Cell Biol, 6, 75–82.
6.
BullejosM., BowlesJ., and KoopmanP. (2002). Extensive vascularization of developing mouse ovaries revealed by caveolin-1 expression. Dev Dyn, 225, 95–99.
7.
Chamier-GliszczyńskaA., BrązertM., Sujka-KordowskaP., PopisM., OżegowskaK., StefańskaK., et al. (2018). Genes involved in angiogenesis and circulatory system development are differentially expressed in porcine epithelial oviductal cells during long-term primary in vitro culture–a transcriptomic study. Med J Cell Biol, 6, 163–173.
8.
EricksonG.F., NakataniA., LingN., and ShimasakiS. (1992). Localization of insulin-like growth factor-binding protein-5 messenger ribonucleic acid in rat ovaries during the estrous cycle. Endocrinology, 130, 1867–1878.
9.
FirthS.M., and BaxterR.C. (2002). Cellular actions of the insulin-like growth factor binding proteins. Endocr Rev, 23, 824–854.
10.
HayashiK.G., UshizawaK., HosoeM., and TakahashiT. (2010). Differential genome-wide gene expression profiling of bovine largest and second-largest follicles: identification of genes associated with growth of dominant follicles. Reprod Biol Endocrinol, 8, 11.
11.
HondaT., FujiwaraH., YoshiokaS., YamadaS., NakayamaT., EgawaM., et al. (2004). Laminin and fibronectin concentrations of the follicular fluid correlate with granulosa cell luteinization and oocyte quality. Reprod Med Biol, 3, 43–49.
12.
HuetC., MongetP., PisseletC., and MonniauxD. (1997). Changes in extracellular matrix components and steroidogenic enzymes during growth and atresia of antral ovarian follicles in the sheep. Biol Reprod, 56, 1025–1034.
13.
InoueO., Suzuki-InoueK., DeanW.L., FramptonJ., and WatsonS.P. (2003). Integrin α2β1mediates outside-in regulation of platelet spreading on collagen through activation of Src kinases and PLCγ2. J Cell Biol, 160, 769–780.
14.
JankowskiM., Dyszkiewicz-KonwińskaM., BudnaJ., HuangY., KnapS., BryjaA., et al. (2018). Does migrative and proliferative capability of epithelial cells reflect cellular developmental competence?. Med J Cell Biol, 6, 1–7.
15.
KamangarB.B., GabillardJ.-C., and BobeJ. (2006). Insulin-like growth factor-binding protein (IGFBP)-1, -2, -3, -4, -5, and -6 and IGFBP-related protein 1 during rainbow trout postvitellogenesis and oocyte maturation: molecular characterization, expression profiles, and hormonal regulation. Endocrinology, 147, 2399–2410.
16.
KrancW., BrązertM., OżegowskaK., Budna-TukanJ., CelichowskiP., JankowskiM., et al. (2018a). Response to abiotic and organic substances stimulation belongs to ontologic groups significantly up-regulated in porcine immature oocytes. Med J Cell Biol, 6, 91–100.
17.
KrancW., BrązertM., OżegowskaK., NawrockiM., BudnaJ., CelichowskiP., et al. (2017). Expression profile of genes regulating steroid biosynthesis and metabolism in human ovarian granulosa cells—a primary culture approach. Int J Mol Sci, 18, 2673.
18.
KrancW., JankowskiM., BudnaJ., CelichowskiP., KhozmiR., BryjaA., et al. (2018b). Amino acids metabolism and degradation is regulated during porcine oviductal epithelial cells (OECs) primary culture in vitro–a signaling pathways activation approach. Med J Cell Biol, 6, 18–26.
19.
Le BellegoF., PisseletC., HuetC., MongetP., and MonniauxD. (2002). Laminin-α6β1 integrin interaction enhances survival and proliferation and modulates steroidogenesis of ovine granulosa cells. J Endocrinol, 172, 45–59.
20.
LiH., YuT., MaY., and WangH. (2017). Expression of Laminin gene family in porcine pluripotent stem cells. Sheng Wu Gong Cheng Xue Bao Aug, 25, 1204–1314.
21.
MatzukM.M., BurnsK.H., ViveirosM.M., and EppigJ.J. (2002). Intercellular communication in the mammalian ovary: oocytes carry the conversation. Science, 296, 2178–2180.
22.
MohanS., NakaoY., HondaY., LandaleE., LeserU., DonyC., et al. (1995). Studies on the mechanisms by which insulin-like growth factor (IGF) binding protein-4 (IGFBP-4) and IGFBP-5 modulate IGF actions in bone cells. J Biol Chem, 270, 20424–20431.
23.
MorleyP., ArmstrongD.T., and Gore-LangtonR.E. (1987). Adhesion and differentiation of cultured rat granulosa cells: role of fibronectin. Am J Physiol, 253, C625–C632.
24.
MuroA.F., ChauhanA.K., GajovicS., IaconcigA., PorroF., StantaG., et al. (2003). Regulated splicing of the fibronectin EDA exon is essential for proper skin wound healing and normal lifespan. J Cell Biol, 162, 149–160.
25.
NawrockiM.J., CelichowskiP., JankowskiM., KrancW., BryjaA., Borys-WójcikS., et al. (2018). Ontology groups representing angiogenesis and blood vessels development are highly up-regulated during porcine oviductal epithelial cells long-term real-time proliferation–a primary cell culture approach. Med J Cell Biol, 6, 186–194.
26.
RazaniB., CombsT.P., WangX.B., FrankP.G., ParkD.S., RussellR.G., et al. (2002). Caveolin-1-deficient mice are lean, resistant to diet-induced obesity, and show hypertriglyceridemia with adipocyte abnormalities. J Biol Chem, 277, 8635–8647.
27.
ReisA., ChangH.Y., LevasseurM., and JonesK.T. (2006). APCcdh1Activity in mouse oocytes prevents entry into the first meiotic division. Nat Cell Biol, 8, 539–540.
28.
RiosH., KoushikS.V., WangH., WangJ., ZhouH.-M., LindsleyA., et al. (2005). Periostin null mice exhibit dwarfism, incisor enamel defects, and an early-onset periodontal disease-like phenotype. Mol Cell Biol, 25, 11131–11144.
29.
RodgersR.J., van WezelI.L., Irving-RodgersH.F., LavranosT.C., IrvineC.M., and KrupaM. (1999). Roles of extracellular matrix in follicular development. J Reprod Fertil Suppl, 54, 343–352.
30.
RybskaM., KnapS., JankowskiM., JesetaM., BukowskaD., AntosikP., et al. (2018a). Characteristic of factors influencing the proper course of folliculogenesis in mammals. Med J Cell Biol, 6, 33–38.
31.
RybskaM., KnapS., JankowskiM., JesetaM., BukowskaD., AntosikP., et al. (2018b). Cytoplasmic and nuclear maturation of oocytes in mammals–living in the shadow of cells developmental capability. Med J Cell Biol, 6, 13–17.
32.
ShiR., SunJ., SunQ., ZhangQ., XiaW., DongG., et al. (2016). Upregulation of FAM83D promotes malignant phenotypes of lung adenocarcinoma by regulating cell cycle. Am J Cancer Res, 6, 2587–2598.
33.
StefańskaK., Chamier-GliszczyńskaA., JankowskiM., CelichowskiP., KulusM., RojewskaM., et al. (2018). Epithelium morphogenesis and oviduct development are regulated by significant increase of expression of genes after long-term in vitro primary culture–a microarray assays. Med J Cell Biol, 6, 195–204.
34.
ThériaultB.L., PajovicS., BernardiniM.Q., ShawP.A., and GallieB.L. (2012). Kinesin family member 14: an independent prognostic marker and potential therapeutic target for ovarian cancer. Int J Cancer, 130, 1844–1854.
35.
UyarA., TorrealdayS., and SeliE. (2013). Cumulus and granulosa cell markers of oocyte and embryo quality. Fertil Steril, 99, 979–997.
36.
WalterW., Sánchez-CaboF., and RicoteM. (2015). GOplot: an R package for visually combining expression data with functional analysis: Bioinformatics. 31, 2912–2914.
37.
WingerQ.A. (1997). Bovine oviductal and embryonic insulin-like growth factor binding proteins: possible regulators of “embryotrophic”. insulin-like growth factor circuits. Biol Reprod, 56, 1415–1423.
38.
WuJ., EmeryB.R., and CarrellD.T. (2001). In vitro growth, maturation, fertilization, and embryonic development of oocytes from porcine preantral follicles. Biol Reprod, 64, 375–381.
39.
YamadaS., FujiwaraH., HondaT., HiguchiT., NakayamaT., InoueT., et al. (1999). Human granulosa cells express integrin alpha2 and collagen type IV: possible involvement of collagen type IV in granulosa cell luteinization. Mol Hum Reprod, 5, 07–617.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
