Norepinephrine plays a central role in several fear-related disorders, such as generalized anxiety disorder (GAD). Pharmacological studies in humans and animals have shown that fear-related behaviors can be regulated by the systemic application of noradrenergic drugs. However, individual differences in trait anxiety are often overlooked when studying the effects of not only noradrenergic drugs, but other compounds. In the present study we examined the effects of yohimbine, an alpha2-adrenergic receptor antagonist, in two lines of rats bred for high and low freezing responses to contextual cues previously associated with footshocks (Carioca high- and low-conditioned freezing rats - CHF and CLF, respectively). We found that systemic administration of yohimbine on the second day of contextual fear conditioning (test session) significantly decreased the freezing responses of CHF females, but not CHF males. Yet, yohimbine treatment induced a significant increase in freezing behavior of both male and female CLF rats. Similar results were observed when groups were re-exposed to the same conditioning chamber 6 days later. Our findings indicate that while yohimbine leads to anxiolytic effects on CHF rats, it has an anxiogenic effect on CLF ones. However, such effect was more evident in females than in males. Our findings point to the role of norepinephrine in regulating and mediating fear responses in different anxiety traits. Furthermore, our findings also underscore the relevance of using both sexes in behavioral and pharmacological studies using animal models of anxiety disorders.