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Previous empirical studies have demonstrated a complex, even contradictory, picture of the relationship between teacher collaboration and teacher autonomy. The current study adds new knowledge about this relationship by employing a rigorous, multilevel cross-lagged panel model based on three years of longitudinal data from the Study of Instructional Improvement. With data collected from 644 school leaders and 5,478 teachers in 114 schools, our results demonstrated that teacher collaboration significantly influences future pedagogical autonomy but has no effect on future content autonomy, while neither content nor pedagogical autonomy demonstrated an influence on teacher collaboration. We discuss explanations for our findings and implications of the study for educational practice and future research.