Testes and ovaries undergo sex-specific morphogenetic changes and adopt strikingly different morphologies, despite the fact that both arise from a common precursor, the bipotential gonad. Previous studies showed that recruitment of vasculature is critical for testis patterning. However, vasculature is not recruited into the early ovary. Peripheral innervation is involved in patterning development of many organs but has been given little attention in gonad development. In this study, we show that while innervation in the male reproductive complex is restricted to the epididymis and vas deferens and never invades the interior of the testis, neural crestderived innervation invades the interior of the ovary around E16.5. Individual neural crest cells colonize the ovary, differentiate into neurons and glia, and form a dense neural network within the ovarian medulla. Using a sex-reversing mutant mouse line, we show that innervation is specific to ovary development, is not dependent on the genetic sex of gonadal or neural crest cells, and may be blocked by repressive guidance signals elevated in the male pathway. This study reveals another aspect of sexually dimorphic gonad development, establishes a precise timeline and structure of ovarian innervation, and raises many questions for future research. organogenesis | testis | ovary | neural crest | innervation T he development of the testis and ovary is a unique model of organogenesis in which two distinct organs arise from the same gonadal precursor tissue, the bipotential gonad. At the stage of sex determination, mouse embryonic day 11.5 (E11.5), bipotential gonads commit to a sex-specific fate and develop into testes in XY embryos and ovaries in XX embryos. The striking morphological dimorphism evident between testes and ovaries is initiated by intrinsic pathways controlled by the sex determination cascade and, at least in the case of the testis, propagated by differential recruitment of extrinsic components involved in patterning the organ.An interaction between the male pathway and the vasculature instructs testis patterning. Vasculature migrates from the mesonephros into the testis based on signals initiated by the male pathway (1). Live imaging showed that endothelial cells first migrate to form the coelomic artery at the surface of the testis, then branch and induce proliferation of the surrounding mesenchyme, which separates Sertoli cells into clearly defined testis cords (2, 3). Disruption of endothelial migration led to severe defects in testis morphogenesis and cord formation (2, 4). These results demonstrated the essential role of endothelial cell migration in patterning the testis. However, recruitment of external vasculature did not occur into the ovary during the stages investigated in these studies (1, 3), thus seemed unlikely to be involved in early patterning events of the ovary, such as germ cell cyst formation and breakdown to form primordial follicles (5, 6). The factors regulating ovary patterning remain largely unknown.Peripheral innervation is an...