Palynological analyses are used in conjunction with oxygen isotopes and Mg/Ca ratios in foraminifers in order to document the response of dinoflagellate cysts (dinocysts) assemblages to changing climate conditions in the northeastern Gulf of Mexico over the Holocene and late Pleistocene. During MIS 6, but also during the cooler phases of MIS 5, Impagidinium species and Operculodinium centrocarpum were dominating the assemblages. By contrast, during the last interglacial (LIG) and the Holocene, assemblages were mainly composed of Spiniferites taxa and characterized by high relative abundance of Spiniferites mirabilis-hyperacanthus, Operculodinium israelianum and/or Polysphaeridium zoharyi. These two periods exhibit ~ 1-2 °C difference in temperature as inferred from Mg/Ca ratios and show significantly distinct assemblages, with higher percentages of S. mirabilis during the LIG and higher percentages of P. zoharyi during the Holocene. This likely denotes important differences in the hydrogeographical conditions (e.g. surface circulation, bathymetric configuration) between the present and last interglacial. The importance of environmental parameters other than temperature and salinity for dinocyst assemblage dynamics is furthermore illustrated.