This study investigated maturation and spawning of Patagonian toothfish Dissostichus eleginoides in the Heard Island and McDonald Islands (HIMI) fishery on the Kerguelen Plateau in the Indian Sector of the Southern Ocean based on gonads and otoliths collected between years 2004–2015, and using histological analyses and calibration of macroscopic staging criteria. Dissostichus eleginoides at HIMI spawn throughout the late autumn and winter months of May–August, and spawning activity is concentrated on slopes along the west and south of the plateau around HIMI at depths of 1500–1900m. Comparison between histological analyses and macroscopic gonad staging indicated that many fish that had spawned, as indicated by the presence of post-ovulatory follicles, returned to a resting stage which was macroscopically indistinguishable from maturing fish. Furthermore, the occurrence of females of all size classes with low gonadosomatic index and low macroscopic gonad stage during the spawning season suggested that a proportion of mature females did not spawn every year. Age-at-maturity estimates, obtained based on the assumption that fish of macroscopic stages ≥ 2 were mature, decreased between the 2004–2009 and 2010–2015 periods for both sexes. However, the magnitude of this temporal variation in age-at-maturity varied between gear types and fishing depths, and variable sampling regimes likely influenced these variations. This study highlights the importance of correct interpretation of macroscopic gonad stages, and understanding the influence of fishery operations on estimations of life-history parameters.