Transcriptome sequencing analysis of porcine granulosa cells treated with an anti-inhibin antibody.

Inhibin can regulate granulosa cell proliferation and function via direct action on granulosa cells, or indirectly through stimulation of pituitary follicle-stimulating hormone secretion. Thus far, it has not been possible to unravel or formulate the chain of molecular events that lead to enhanced granulosa cell proliferation and function using conventional gene expression analysis. The aim of this study was to examine the biological effects of immuno-neutralization of inhibin bioactivity in porcine granulosa cells using transcriptome profiling by the RNA-seq technology. Treatment of granulosa cells with anti-inhibin α subunit antibodies increased both cell proliferation and estradiol secretion. Data revealed by RNA sequencing were subjected to bioinformatic analysis. The results showed that a total of 476 genes, including 27 novel genes, were differentially expressed in anti- inhibin antibody-treated granulosa cells compared to untreated granulosa cells. RNA sequencing data were validated by qRT-PCR which confirmed differential expression (upregulation and downregulation) of eighteen of twenty selected genes A total of 476 differentially expressed genes were enriched in processes such as matrix remodeling, chemokine activity, protein binding, and structural molecular activities, and which could be related to granulosa cell proliferation, estradiol synthesis, and ovarian follicle growth. In particular, the data emphasized the importance of extracellular matrix remodeling and the involvement of chemokines in enhanced granulosa cell function, which are important features of ovarian follicle growth, development, maturation, and ovulation. This study provided a new level of understanding of enhanced granulosa cell function and ovarian follicle development achieved through immuno-neutralization of endogenous inhibin bioactivity.