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Separated throughout most of the Cenozoic era, North and South America were joined during the mid-Pliocene when the uplift of Panama formed a land bridge between these two continents. The fossil record indicates that this connection allowed an unprecedented degree of inter-continental exchange to occur between unique, previously isolated biotic assemblages, a phenomenon now recognized as the “Great American Biotic Interchange”. However, a relatively poor avian fossil record has prevented our understanding the role of the land bridge in shaping New World avian communities. To address the question of avian participation in the GABI, we compiled 64 avian phylogenetic studies and applied a relaxed molecular clock to estimate the timing of trans-isthmus diversification events. Here, we show that a significant pulse of avian interchange occurred in concert with the isthmus uplift. The avian exchange was temporally consistent with the well understood mammalian interchange, despite the presumed greater vagility of birds. Birds inhabiting a variety of habitats and elevational zones responded to the newly available corridor. Within the tropics, exchange was equal in both directions although between extratropical and tropical regions it was not. Avian lineages with Nearctic origins have repeatedly invaded the tropics and radiated throughout South America; whereas, lineages with South American tropical origins remain largely restricted to the confines of the Neotropical region. This previously unrecognized pattern of asymmetric niche conservatism may represent an important and underappreciated contributor to the latitude diversity gradient.