Granulin precursor gene: a sex steroid-inducible gene involved in sexual differentiation of the rat brain

The mechanisms of sexual differentiation of the brain by sex steroids seem to be conserved throughout the mammalian species, although there may be some species differences. In rats, sex-dependent differentiation of the brain occurs in a sex steroid-dependent manner during the perinatal period known as the critical period. Androgen exposure during the perinatal period results in the development of structural and functional sexually dimorphic characteristics in the brain; the absence of testicular androgen leads the central nervous system to develop passively in a primarily female fashion, while the presence of androgen induces the masculinization of the brain. We attempted to characterize sex steroid-inducible genes that are involved in the sexually dimorphic function of the brain. Following the cDNA subtraction between hypothalami of 5-day-old intact and neonatally androgenized female rats, a granulin (grn) precursor gene was identified. The grn gene encodes a 6-kDa polypeptide known as a growth modulating factor of epithelial cells in vitro. Exogenous estrogen, as well as androgen, induced grn gene expression in the neonatal hypothalamus. In the brain of a 5-day-old male rat, grn mRNA was expressed in the ventromedial hypothalamic nucleus and the arcuate nucleus of the hypothalamus. Throughout the critical period for sexual differentiation of the brain, grn gene expression remained high in males, while in females it gradually decreased. Antisense oligodeoxynucleotide (ODN) complementary to grn mRNA was synthesized and infused into the third ventricle of male rats at 2 days of age. Two different control treatments were used; the first consisted of a control sequence ODN that had virtually no homology to known mRNAs, and the second consisted of vehicle alone. After maturation, the subject animals that were treated with antisense ODN of grn displayed significantly lower scores than the control males in various parameters assessing sexual behavior, i.e., mount, intromission, and ejaculation. The present results suggest that the grn gene, the expression of which is induced by sex steroids in the neonatal hypothalamus, plays a crucial role in the functional masculinization of the rat brain.