Identification of Differentially Expressed Genes in Bovine Follicular Cystic Ovaries

Abstract

Follicular cystic ovary (FCO) is one of the most frequently diagnosed ovarian diseases and is a major cause of reproductive failure in mammalian species. However, the mechanism by which FCO is induced remains unclear. Genetic alterations which affect the functioning of many kinds of cells and/or tissues could be present in cystic ovaries. In this study, we performed a comparison analysis of gene expression in order to identify new molecules useful in discrimination of bovine FCO with follicular cystic follicles (FCFs). Normal follicles and FCFs were classified based on their sizes (5 to 10 mm and ≥25 mm). These follicles had granulosa cell layer and theca interna and the hormone 17β-estradiol (E2)/ progesterone (P4) ratio in follicles was greater than one. Perifollicular regions including follicles were used for the preparation of RNA or protein. Differentially expressed genes (DEG) that showed greater than a 2-fold change in expression were screened by the annealing control primer (ACP)-based PCR method using GeneFishing TM DEG kits in bovine normal follicles and FCFs. We identified two DEGs in the FCFs: ribosomal protein L15 (RPL15) and microtubule-associated protein 1B (MAP1B) based on BLAST searches of the NCBI GenBank. Consistent with the ACP analysis, semi-quantitative PCR data and Western blot analyses revealed an up-regulation of RPL15 and a down-regulation of MAP1B in FCFs. These results suggest that RPL15 and MAP1B may be involved in the regulation of pathological processes in bovine FCOs and may help to establish a bovine gene data-base for the discrimination of FCOs from normal ovaries.

Comparative genomics between fly, mouse, and cattle identifies genes associated with sire conception rate

“…However, further studies are required to validate these effects. Recently, the expression of MAP1B was found to be downregulated in follicular cystic follicles compared with normal follicles, suggesting that alteration in MAP1B expression may be involved in reproductive failure in cattle (Choe et al, 2010). These results again demonstrate the significance of both parental genomes to embryonic development and fertility.…”

Blastocyst implantation failure relates to impaired translational machinery gene expression

“…The mRNA of Rpl15 decreases during bovine oocytes meiotic maturation and increases in the morula and blastocyst stages of bovine embryogenesis (Bettegowda et al 2006). This gene is upregulated in bovine follicular cystic ovaries and is probably responsible for the delayed regression with persistent follicle growth (Choe et al 2010). We further suggest that Cx43 depletion modulate also ribosomal proteins expression affecting acquisition of developmental competence by the oocyte as well.…”

Aquaporin 4 expression is downregulated in large bovine ovarian follicles

“…The experimental materials and methods were used similarly to previous studies conducted in this laboratory (Siregar et al, 2019;Choe et al, 2010).…”

“…The ovarian follicles were classified into sizes as small (< 10 mm, 5 to 10 mm) and large (> 25 mm in diameter). The isolation procedures were performed as previously described (Choe et al, 2010). Briefly, follicular fluid (FF) was carefully aspirated from small and large follicles with a 10 mL syringe fitted with an 18 or 23 gauge needle; the fluid was centrifuged at 1,750 × g for 10 min, and the resultant pellets were used for identification of AQP4 expression.…”