Dispersal, a critical factor in the viability of marine organisms, is strongly influenced by the physical dynamics of the ocean. Fronts, as a common phenomenon in ocean dynamics, can theoretically impact the dispersal of organisms and ultimately cause spatial homogeneity or heterogeneity of marine communities. However, there is relatively limited understanding concerning whether frontal formation hinders the connectivity between communities and thus results in differences in community structure. To verify whether the formation of fronts caused differences between communities, 60 eDNA samples and their environmental parameters were collected from 32 sites in the Yangtze River estuary and its adjacent area, and four groups, the Yangtze River front-inner group (YRI), the Yangtze River front-outer group (YRO), the Zhejiang costa front-inner group (ZCI), and the Zhejiang coast front-outer group (ZCO), were compared to examine the differences in fish communities between the sides of the front. The results showed that there was a significant difference between YRI and YRO, suggesting that the front affected the community structure. There was no significant difference in fish communities between ZCI and ZCO, indicating that fronts could only hinder dispersal at high intensities. The difference between YRI and YRO possibly resulted from dramatic changes in temperature, salinity, and turbidity related to the front. Fronts also separated water masses with different environmental characteristics and thereby attracted fish to their optimal habitats. Overall, fronts appear to be ecologically significant for fish dispersal and community differentiation.