The Younger Dryas (YD) climatic reversal (12.86-11.65 cal ky BP), especially the warming initiated at ∼12.6 cal ky BP, and the associated vegetation changes have been proposed as past analogs to forecast the potential vegetation responses to future global warming. In this paper, we applied this model to highland and midland Neotropical localities. We used pollen analysis of lake sediments to record vegetation responses to YD climatic changes, which are reconstructed from independent paleoclimatic proxies such as the Mg/Ca ratio on foraminiferal tests and Equilibrium Line Altitude (ELA) for paleotemperature, and grayscale density and Titanium content for paleoprecipitation. Paleoclimatic reconstructions at both highlands and midlands showed a clear YD signal with a conspicuous warming extending into the early Holocene. A small percentage of taxa resulted to be sensitive to these YD climate changes. Response lags were negligible at the resolution of the study. However, changes in the sensitive taxa were relevant enough to determine changes in biodiversity and taxonomic composition. Highland vegetation experienced mainly intra-community reorganizations, whereas midland vegetation underwent major changes leading to community substitutions. This was explained in terms of threshold-crossing non-linear responses in which the coupling of climatic and other forcings (fire) was proposed as the main driving mechanism. Paleoecology provides meaningful insights on the responses of highland and midland Neotropical vegetation to the YD climatic reversal. Biotic responses at both individual (species) and collective (assemblage) levels showed patterns and processes of vegetation change useful to understand its ecological dynamics, as well as the mechanisms and external drivers involved. The use of paleoecological methods to document the biotic responses to the YD climate shifts can be useful to help forecasting the potential consequences of future global warming. Due to its quasi-global character, the YD reversal emerges as a well suited candidate for providing useful insights of global scope by analyzing the corresponding biotic responses virtually at any geographical and biological setting.