Functional pain disorders disproportionately impact females, but most pain research in animals has been conducted in males. While there are anatomical and pharmacological sexual dimorphisms in brainstem pain-modulation circuits, the physiology of pain-modulating neurons that comprise a major functional output, the rostral ventromedial medulla (RVM), has not been explored in female animals. The goal of this study was to identify and characterize the activity of RVM cells in female, compared to male, rats. ON- and OFF-cells were identified within the RVM in females, with firing properties comparable to those described in males. In addition, both ON- and OFF-cells exhibited a sensitized response to somatic stimuli in females subjected to persistent inflammation, and both ON- and OFF-cells responded to systemically administered morphine at a dose sufficient to produce behavioral antinociception. These data demonstrate that the ON-/OFF-cell framework originally defined in males is also present in females, and that as in males, these neurons are recruited in females in persistent inflammation and by systemically administered morphine. Importantly, this work establishes a foundation for the use of female animals in studies of RVM and descending control.