Effects of FSH on testicular mRNA transcript levels in the hypogonadal mouse. Mus musculus

In the normal mouse the pituitary gonadotrophins determine development, maturation and physiological regulation of the testis with follicle-stimulating hormone (FSH) activating the Sertoli cell and luteinising hormone (LH) the Leydig cell. To look at the potential interaction of cell types within the testis following hormone stimulation we have investigated the effect of rFSH on testicular gene expression in the hypogonadal (hpg) male mouse. Due to a deletion in the gene encoding gonadotrophin-releasing hormone (GnRH), FSH and LH levels are at the lower limit of detection in the circulation and mice remain in a pre-pubertal state throughout life unless given exogenous hormone. Here we have addressed important question of changes in the global gene expression profile of testicular tissue following hormone stimulation (rFSH) for 12, 24 and 72h following injections compared to control untreated hpg males of the same age. Keywords: Hormone Response, Testicular gene expression Profile Overall design: Adult 10 week old hpg males were injected sc with 8 I.U. of rFSH (Gonal F, Serono Pharmaceuticals) in 0.2ml of PBS (Sigma,UK) every 12 h for 12, 24 and 72 h. Mice were killed one hour after the last injection by cervical dislocation, testes dissected out, weighed and snap frozen in liquid nitrogen. Tissue was stored at -70oC until use. RNA was extracted using Qiagen RNeasy Lipid Tissue Mini kit (Qiagen, Valencia, CA) according to the manufacturer’s instructions and quantified on a NanoDrop ND-1000 (NanoDrop Wilmington, DE). RNA quality was checked using the Agilent bioanalyzer 2100 (Agilent, Santa Clara, CA). Testicular gene expression was examined in four individual mice at 12 and 72 h and 3 mice at 24h using high-density oligonucleotide mouse MOE430A arrays (Affymetrix, Santa Clara, CA), comparison was made with a group of 7 untreated hpg males of the same age.