Purpose/Hypothesis: Physical therapy specialties were developed in order to meet the specialized health needs of diverse populations. Designation as a board-certified specialist requires additional training that often includes advanced gross anatomy knowledge and application. Although anatomy educational research is ongoing in many health-related professions, few empirical findings have been produced regarding the translation of anatomy knowledge from a basic science to a tool of specialty practice, none of which are in the profession of physical therapy. The objective of this study was to determine whether board-certified clinical specialist physical therapists possess dissimilar gross anatomy knowledge when compared to other board-specialists and non-specialist physical therapists. Specifically, two hypotheses were investigated: 1) board-certified specialists would show differing anatomy knowledge compared to those who are not board-certified; 2) differences in anatomy content knowledge exist among board-certified specialists based on specialty area. Number of Subjects: 3,535 Materials and Methods: The 3,535 sample was composed of board-certified physical therapists (n=617) and non-certified physical therapists (n= 2,918), with 65.7% female, 33.3% male, and 0.1% other. Mean years of practice was 13.48 (SD= 11.59). Participants completed a questionnaire of 30 items demonstrating their anatomy knowledge in six (6) regional anatomy areas: 1) upper extremity; 2) lower extremity; 3) thorax and abdomen; 4) pelvis; 5) spine; and 6) head. Each regional area contained five (5) questions based in systemic subsets: 1) joints and osteology; 2) muscular anatomy; 3) nervous system; 4) vasculature; and 5) special areas. Results: Results revealed board-certified specialists (M=17.85) exhibited statistically significant greater anatomy knowledge on the questionnaire compared to non-certified specialists (M=16.09). Board-certified physical therapists across the eight specialty areas exhibited varying regional anatomy knowledge. 9.2% of the variance in questionnaire score was explained by the specific specialty area of the board-certified physical therapists. Conclusions: This study provided empirical findings consistent with the hypothesis that board-certified specialists showed diverse knowledge of anatomy content. Significant findings included the following: 1) physical therapists who are board-certified specialists showed a greater amount of anatomy knowledge than physical therapists who were not board-certified specialists; 2) board-certified specialists demonstrated diverse knowledge in regional anatomy topics across the eight specialty areas. Clinical Relevance: This study was a preliminary step in investigating the extent to which anatomy knowledge plays a role in expert clinical practice for physical therapists. This study prepared a foundation for the longitudinal aim of anatomy curricula and educational advancements for physical therapists to meet the growing professional demands for expert practice.