Gene expression profiling of upregulated mRNAs in granulosa cells of bovine ovulatory follicles following stimulation with hCG

Reproductive Biology and Endocrinology

Research

Jean Sirois¹  Kalidou Ndiaye¹  Jacques G. Lussier¹  Valérie Lévesque¹  Mame N. Diouf² 

[1] Centre de recherche en reproduction et fertilité, Faculté de médecine vétérinaire, Université de Montréal, 3200 Sicotte, St-Hyacinthe, J2S 2M2, Québec, Canada;Centre de recherche en reproduction et fertilité, Faculté de médecine vétérinaire, Université de Montréal, 3200 Sicotte, St-Hyacinthe, J2S 2M2, Québec, Canada;Institut Sénégalais de Recherches Agricoles (ISRA) Laboratoire National de l′Elevage et de Recherches Vétérinaires (LNERV), BP 2057, Dakar-Hann, Sénégal;

关键词: Ovary;    Follicle;    Ovulation;    Granulosa cells;    Gene expression;   

DOI  :  10.1186/s12958-017-0306-x

 received in 2017-09-12, accepted in 2017-10-16,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundOvulation and luteinization of follicles are complex biological processes initiated by the preovulatory luteinizing hormone surge. The objective of this study was to identify genes that are differentially expressed in bovine granulosa cells (GC) of ovulatory follicles.MethodsGranulosa cells were collected during the first follicular wave of the bovine estrous cycle from dominant follicles (DF) and from ovulatory follicles (OF) obtained 24 h following injection of human chorionic gonadotropin (hCG). A granulosa cell subtracted cDNA library (OF-DF) was generated using suppression subtractive hybridization and screened.ResultsDetection of genes known to be upregulated in bovine GC during ovulation, such as ADAMTS1, CAV1, EGR1, MMP1, PLAT, PLA2G4A, PTGES, PTGS2, RGS2, TIMP1, TNFAIP6 and VNN2 validated the physiological model and analytical techniques used. For a subset of genes that were identified for the first time, gene expression profiles were further compared by semiquantitative RT-PCR in follicles obtained at different developmental stages. Results confirmed an induction or upregulation of the respective mRNAs in GC of OF 24 h after hCG-injection compared with those of DF for the following genes: ADAMTS9, ARAF, CAPN2, CRISPLD2, FKBP5, GFPT2, KIT, KITLG, L3MBLT3, MRO, NUDT10, NUDT11, P4HA3, POSTN, PSAP, RBP1, SAT1, SDC4, TIMP2, TNC and USP53. In bovine GC, CRISPLD2 and POSTN mRNA were found as full-length transcript whereas L3MBLT3 mRNA was alternatively spliced resulting in a truncated protein missing the carboxy-terminal end amino acids, 774KNSHNEL780. Conversely, L3MBLT3 is expressed as a full-length mRNA in a bovine endometrial cell line. The 774KNSHNEL780 sequence is well conserved in all mammalian species and follows a SAM domain known to confer protein/protein interactions, which suggest a key function for these amino acids in the epigenetic control of gene expression.ConclusionsWe conclude that we have identified novel genes that are upregulated by hCG in bovine GC of OF, thereby providing novel insight into peri-ovulatory regulation of genes that contribute to ovulation and/or luteinization processes.

【 授权许可】

CC BY   
© The Author(s). 2017