This doctoral thesis presents the results of the pioneering archaeological investigation conducted in the Northern Mexican Highlands with the aim to evaluate the existing indicators of the earliest human occupations at the end of the Pleistocene and discover new evidence of ancient cultural manifestations through a systematic exploration of an endorheic basic in the Zacatecas desert, a region never studied before. An exhaustive survey and analysis of the available literature on Mexican prehistory establishes the weak points of the local paradigms, differentiating between academic myths and objective realities. A complete historiography of the topic of the earliest humans in Mexico has been achieved, for the first time. The study of several collections of flaked stone artefacts, in different cities in Mexico, show new indicators of the presence of bearers of the Late Paleoamerican cultures, in regions where their presence had been weakly confirmed. The most important part of the research consisted in fieldwork realised during two long seasons; the first one dedicated to the surface explorations and the second one to excavations. Thirty-five new archaeological sites were discovered in the first phase, most of them open campsites reminiscent of hunter-gatherer societies, with a richness of stone artefacts on their surface. They indicate a long cultural sequence, going from the Late Pleistocene to the Late Holocene and the historic periods. Four sites were further studied by fourteen test excavation units: Dunas de Milpa Grande, San José de las Grutas, the Chiquihuite Cave and Ojo de Agua. Two new archaeological cultures were identified, one at Dunas (an interesting assemblage of limestone and basalt flaked stone tools) and another one at San José (a limestone concave-based points complex). First indicators of ʻolder than Clovisʼ human presence have also been obtained. The palaeoenvironmental data provide a preliminary reconstruction of the Late Pleistocene-Early Holocene landscape of the basin, based on geology, extinct fauna, phytolith and mollusc analyses. Radiocarbon and OSL results support a first cultural and paleoclimatic model for the study area. This investigation also discovered the first case of a “black mat” in Mexico: a black layer of sediment deposited under specific environmental conditions during the Younger Dryas cooling event.