Parkinson's disease (PD) is a progressive neurodegenerative disorder characterized by loss of dopamine (DA) neurons in the substantia nigra pars compacta (SNC). Most epidemiologic studies have demonstrated that PD has a higher prevalence in males than in females. Both hormones and genetic factors have been considered to be contributors to this phenomenon. In the present study, we used primary cultures of ventral mesencephalic (VM) neurons from E13.5 Balb/C mice to investigate whether there were any gender differences in gene expression and cell sensitivity to oxidative stress in sex segregated cultures. We also investigated the role of SRY, the sex-determining region on the Y chromosome, and the female hormone estrogen in the gender dimorphism. We measured the expression levels of genes that previously were thought to be related to PD or DA neuron development and functions by real-time PCR, and found six of them, ATP13A2, ERβ, MAO-A, D2, DAT, and Pitx3, showing significantly differential expression between males and females in the normal physiological state or under stress conditions. Furthermore, we demonstrated that male VM neurons are more vulnerable than female neurons to rotenone-induced cytotoxicity and that 17β-estrogen has a moderate protective effect in both male and female VM neurons. Moreover, we document that SRY can upregulate monoamine oxidase A and downregulate estrogen receptor-β, and SRY-overexpressing N2A cells enhance the resistance to oxidative stress-induced cell injury. Our results suggest that gender indeed influences several PD-related gene expressions in VM neurons, and SRY and estrogen are involved in the different responses to oxidative stress in culture. © 2011 Wiley Periodicals, Inc.