Abstract The current study is an attempt to explore the influence of gender on the prosodic behavior of Iranian male and female speakers in same-sex and cross-sex situations. In this research, it is hypothesized that the gender of the speakers is an influential factor to create a difference in their linguistic behavior rather than other factors such as formality of the context. Talks made by pairs of Iranian male and female speakers whose speech were totally formal were examined from an acoustical point of view by measuring the fundamental frequency of the speakers’ speech. One-way ANOVA was run on the data and the results indicated that women had lower pitch in their cross-sex talks compared with their talks in same-sex situations. However, men showed to have higher pitch when their talk occurred in a cross-sex situation, compared with the situations where they talked with other men. This proves that men and women do not react linguistically similarly in the same situations; and thus, they endeavor to insist on linguistic differences resulted from gender differences.