This dissertation argues that Venetian space in the sixteenth century was embedded with various boundaries that individuals challenged and that communities and Venetian secular and ecclesiastical authorities reinforced. The development of Venetian urban space played an essential role in the formation of Venetian civic identity, which in turn was predicated upon the myth of Venice. The time period examined includes the re-establishment of the Roman Inquisition, and the early period of the Inquisition in Venice, which were concomitant with a time of religious and social disruption. Documents of the Venetian government and contemporary diarists offer contextual evidence; however, trials before the Holy Office in Venice, particularly cases involving those accused of witchcraft, inform the greatest portion of this study. Drawing on such evidence, this dissertation challenges the argument that “Venetian” society was cohesive and well balanced. By repurposing common and sacred items to invoke supernatural entities and perform heterodox practices, those accused of witchcraft challenged the Venetian secular and ecclesiastical authorities as they created a competing vision regarding the definition of domestic sacred space. Examination of the neighborhood as a social space reveals boundaries, both real and imagined, and the challenges to the boundaries that those living on the margins of society displayed through the creation of their own communities. Finally, inhabitants’ use of public space and their movement throughout these spaces offers evidence of challenges to boundaries as well as the measures authorities took in re-establishing these boundaries. Ultimately, competing desires for belonging and legitimacy, as well as disagreements over physical, ideological, and social boundaries set Venetian inhabitants and authorities in opposition.