Zooplankton communities were examined for the first time in three different hydrographic regions of the Hudson Bay system (HBS) in early August to early September from 2003 to 2006. Sampling was conducted at 50 stations distributed along different transects located in Hudson Bay (HB), Hudson Strait (HS), and Foxe Basin (FB). Variations in zooplankton biomass, abundance, taxonomic composition, and diversity in relation to environmental variables were studied using multivariate techniques. During all sampling years, the total zooplankton biomass was on average four times lower in HB than in HS and FB. Clustering samples by their relative species compositions revealed no interannual variation in zooplankton community but showed a marked interregional variability between the three regions. Water column stratification explained the greatest proportion (25%) of this spatial variability. According to redundancy analysis (RDA), the zooplankton taxa that contribute most to the separation of the three regions are Microcalanus spp., Oithona similis, Oncaea borealis, Aeginopsis laurentii, Sagitta elegans, Fritillaria sp., and larvae of cnidaria, chaetognatha, and pteropoda in HB; hyperiid amphipods in FB; and Pseudocalanus spp. CI–CV, Calanus glacialis CI–CVI, Calanus finmarchicus CI–CVI, Calanus hyperboreus CV–CVI, Acartia longiremis CI–CV, Metridia longa N3-N6 CI-CIII CVIf, Eukrohnia hamata, larvae of echinodermata, mollusca, cirripedia, appendicularia, and polychaeta in the northwestern and southeastern HS transects. For the HB transect, the RDA analyzed allowed us to distinguish three regions (HB west, central, and east) with different environmental gradients and zooplankton assemblages, in particular higher concentration of Pseudocalanus spp. nauplii and CI–CVI, as well as benthic macrozooplankton and meroplankton larvae in western HB. In HS, Calanoid species (mainly C. finmarchicus and C. glacialis) were mostly observed at the north shore stations associated with the weakly stratified Arctic–North Atlantic waters coming from southwestern Davis Strait (inflow). In general, the RDA models tested among the HBS regions were very consistent with its general surface circulation pattern for summer conditions in terms of environmental variables and distinct zooplankton assemblages. Overall, zooplankton biomass and diversity indices (H′, J′, and S) were lower in the most stratified environment (i.e., HB) than in the deeper (FB) and more dynamic (HS) regions. The results of this work clearly show that the spatial differentiation and structure of the zooplankton communities are strongly influenced by the hydrodynamic conditions in the HBS that, trough their actions on temperature, salinity, stratification, mixing conditions and depth strata, lead to the spatial differentiation of these communities.