Transport infrastructures are a major driver of global landscape change. In many areas, highways were built before environmental impact assessments required the implementation of wildlife crossing structures. A fundamental task in the development of ecological networks is identifying areas where dispersal pathways of vulnerable populations may be blocked or affected by high volume roads. Financial considerations may limit investment, thus it is vital to establish the relative contribution of each road defragmentation location to upholding habitat connectivity and availability. In this paper, recent methodological developments in habitat network analysis, derived from the probability of connectivity index, were refined and used to evaluate the role of each highway defragmentation location in restoring landscape connectivity. The forest protected area network in the Basque country (northern Spain) was selected as an illustrative case study. The proposed approach was able to quantify and effectively account for a combination of factors determining the contribution of each highway defragmentation location to upholding connectivity, which are rarely jointly considered in analyses oriented to support restoration decisions in landscapes affected by the barrier effect of transport infrastructures. The contribution to connectivity of each defragmentation location depended on its topological position in the landscape, on the relative decrease in the effective distance among habitat areas that results from the permeability restoration at that location, on the distance from the defragmentation location to other alternative wildlife crossing structures already existing in the landscape, on the amount of habitat in the areas connected by the linkages that run through the defragmentation location and on the dispersal abilities of the focal species.