We explore the connection between US designated wilderness areas and wildlife with the goal of establishing a research agenda for better understanding this complex relationship. Our research agenda has two components. The first, “wildlife for wilderness,” considers the impact of wildlife on wilderness character. Whereas studies show that wildlife is important in both the perception and actual enhancement of wilderness character, the context and particulars of this relationship have not been evaluated. For instance, is knowing that a rare, native species is present in a wilderness area enough to increase perceptions of naturalness (an important wilderness quality)? Or does the public need to observe the species or its sign (e.g., tracks) for this benefit? The second part of our research agenda, “wilderness for wildlife,” considers the types of research needed to understand the impact of wilderness areas on wildlife and biodiversity conservation. Several studies show the effect of one area being designated wilderness on one wildlife species. Yet, there has been no research that examines how the networks of wilderness areas in the National Wilderness Preservation System (NWPS) are used by a species or a community of species. Furthermore, we found no studies that focused on how the NWPS affects ecological or trophic interactions among species. We hope that by providing a research agenda, we can spur multiple lines of research on the topic of wildlife and wilderness.