Canonical pathways and networks regulated by estrogen in the bovine mammary gland

Many attempts have been made to identify estrogen-responsive genes using high-throughput approaches such as microarray, serial analysis of gene expression (SAGE), and in silico prediction. However, few studies have systematically analyzed regulatory networks and pathways affected by estrogen. In this report, we analyzed transcript profiles obtained from 16 prepubertal heifers in a 2 x 2 factorial experiment, with ovarian status (intact or ovariectomized) as the first factor and estrogen treatment as the second (control or estradiol). After 54 h of estrogen treatment, gene expression was evaluated in the parenchyma and fat pad of the bovine mammary gland using a high-density oligonucleotide microarray. The genes significantly regulated by estrogen were subject to pathway and regulatory network analysis using Ingenuity Pathways Analysis software. Approximately 2,344 genes responded significantly to estrogen treatment. Of these, 1016 genes were influenced by estrogen regardless of tissue or ovarian status, while the remaining genes were significant in one of four specific effects of tissue or ovarian status. The canonical pathways significantly regulated by estrogen (P < 0.05) included protein ubiquitination, G2/M cell cycle control, IGF1 signaling, N-glycan biosynthesis, sterol biosynthesis, and oxidative phosphorylation. A total of 23 regulatory networks were identified as estrogen responsive. The results provide insight into the molecular mechanisms through which estrogen regulates bovine mammary gland growth and development, supporting the concept that interaction between tissues within the mammary gland promotes mammary epithelial growth.