Objectives: Understanding variations in disease presentation in men and women is clinically important as differences may reflect biological and sociocultural factors and have implications for selecting appropriate prevention and treatment strategies. The aim of this study was to investigate clinical and cognitive differences in treatment-seeking people with pathological gambling as a function of gender. Materials and methods: 501 adult subjects (n = 274 [54.7%] females) with DSM-IV pathological gambling presenting for various clinical research trials over a 9-year period were assessed in terms of sociodemographics and clinical characteristics. A subset (n = 77) had also undertaken neuropsychological assessment with the Stop-signal and set-shift tasks. Results: PG in females was associated with significantly worse disease severity, elevated mood and anxiety scores, and history of affective disorders, later age of study presentation, later age of disease onset, and elevated risk of having a first-degree relative with gambling or alcohol problems. These findings were of small effect size (0.20-0.35). Additionally, PG in females was associated with proportionately more non-strategic gambling with medium effect size (0.61). In contrast, PG in males was associated with a significantly greater lifetime history of an alcohol use disorder and any substance use disorder (small effect sizes 0.22-0.38); and slower motoric reaction times (medium effect size, 0.50). Response inhibition and cognitive flexibility were similar between the groups. Conclusions: These data suggest that important differences exist in the features of pathological gambling in women and men. Findings are of considerable relevance to clinicians and in terms of targeted treatments.