Active immunization against GnRH reduces the synthesis of GnRH in male rats

We sought to determine the effects of active anti-GnRH immunization on GnRH synthesis in the hypothalamus. Adult male rats (n = 36) were randomly and equally allocated into three groups: Control (no treatment), surgically castrated, or immunized against 50 μg D-Lys6-GnRH-tandem-dimer peptide conjugated to ovalbumin in Specol adjuvant at 12 week of age (with a booster 8 week later). Blood samples (for antibody titers and hormone concentrations) were collected at 2-week intervals until rats were killed (20 week). Compared with intact controls, immunocastration reduced (P < 0.05) serum concentrations of testosterone, LH, and FSH, and GnRH content in the median eminence, reduced the weight of the hypohysis (P < 0.01), and induced testicular atrophy (suppression of spermatogenesis). Furthermore, mRNA expression of GnRH in the hypothalamus, GnRH receptor, LH-β and FSH-β in the pituitary, LH receptor and FSH receptor in the testes, and genes in sex steroid feedback loops (androgen receptor [AR], kisspeptin encoded gene (Kiss-1), and kisspeptin receptor (GPR54) in the hypothalamus were decreased in immunocastrated rats compared with intact controls (P < 0.05). Similarly, surgical castration reduced GnRH in the median eminence as well as mRNA expression of GnRH, AR, Kiss-1, and GPR54 in the hypothalamus (P < 0.05). We concluded that anti-GnRH immunization in adult rats reduced synthesis of hypothalamic GnRH by decreasing androgen-AR-Kisspeptin-GPR54 signaling pathways, and caused dysfunction of the pituitary-testicular axis, thereby suppressing spermatogenesis, resulting in testicular atrophy.