And estrogen production [17,18]. It was confirmed, that either sex hormones or some endocrine-disrupting chemical substances, like phthalates, acted by means of PPAR and PPAR, leading to the modulation of PPAR activities . In mouse Leydig cells, Gazouli et al.  found the contribution of cholesterol-transporting proteins within the regulation of steroidogenesis by PPAR. In addition, Kowalewski et al.  demonstratedAnimals 2021, 11,3 ofthat the activation of PPAR by its ligand stopped steroidogenesis in these cells. Our recent research showed modifications inside the quantity of lipid droplets plus the ultrastructure of mitochondria in PPAR-blocked immature boar Leydig cells . It is actually worth mentioning that a new generation of PPAR-targeting pharmacological drugs is undergoing clinical trials for the remedy of infertility associated with metabolic disorders which include insulin resistance . Additionally, PPAR ligands are advised to become utilized for the amelioration of preeclampsia during hypertension and inflammation. Moreover, the accomplishment price of the in vitro fertilization method could be enhanced in the coming years by the enrichment from the culture media with PPAR/ ligands . In prostate and testicular cancers, PPAR constitutes a potent target to treat and avoid these diseases . The membrane G-protein coupled estrogen receptor (GPER) could be the initially sensor for the endogenous and environmental action of estrogenic compounds, e.g., BPA (bisphenol A) that triggers different intracellular targets by way of the activation of fast nongenomic mechanisms . Many current research revealed GPER presence and its implication within the testes functions of a variety of animals . In addition, the association of GPER and estrogenrelated receptors or PPAR interaction inside the Fadrozole custom synthesis handle of healthy and tumor testicular cells in rodents and humans was previously demonstrated by us [6,291]. Currently, the GPER antagonist is monitored as a possible therapeutic drug for male infertility therapy . Nonetheless, the mechanisms of both PPAR and GPER action stay unclear plus the potential long-term adverse effects in fertility handle are unknown. Hence, further investigations are urgently needed. The present investigation was 8-Isoprostaglandin F2�� supplier focused on the determination of genes and gene-controlled molecular processes within the immature boar testes soon after PPAR, PPAR or GPER pharmacological antagonist treatment. The global gene expression analysis of differentially regulated transcripts was followed by a cluster analysis as well as a pathway evaluation, respectively. This integrated an evaluation of differentially regulated genes and proteins expressed in testes with a identified morphological status that were partially earlier studied by us [6,281,33]. For the very best of your authors’ understanding, no research has engaged in this sort of approach just before. two. Materials and Techniques 2.1. Tissue Collection and Ex Vivo Culture Within the Republic of Poland, male piglets are routinely castrated inside the early days just after birth to avoid the production in the main testicular androgen, androstenone, which accumulates in fat and results within the improvement on the boar taint. The testes (n = 40) of 7-day-old Polish White Large boars (n = 20) from a breeding farm in Malopolska, Krakow, Poland were obtained during anaesthetized surgical castration of animals. Tissues were transported to the laboratory in Dulbecco’s phosphate-buffered saline (DPBS, SigmaAldrich, St Louis, MO, USA) supplemented with two penicillin-streptomycin option (Invitroge.