We examined how the lipid content and fatty acid composition of walleye (Sander vitreus) ova varied with respect to maternal characteristics, particularly indices of body nutrient reserves, within three spawning populations that varied in maternal age, size, and somatic lipid reserves. We also compared the variability in ova lipid composition among three populations with that observed among spawning years within one of these populations. Maternal characteristics had little influence on ova lipid content. In Lake Ontario, percentages of docosahexaenoic acid (22:6(n-3), DHA) and arachidonic acid (20:4(n-6), AA) increased with maternal length, while those of eicosapentaenoic acid (20:5(n-3), EPA) declined. In Lake Nipissing, maternal age had similar effects on AA and EPA, but not DHA. The Lake Winnipeg population did not conform to the trends of the other two populations, and ova from this population had very low levels of AA. We hypothesize that there are similar selection pressures on the Lake Ontario and Lake Nipissing populations with respect to desirable ova fatty acid profiles. Lake Winnipeg walleye may experience different selection pressures or may be unable to conform to the trends observed in the other two lakes.