Homo sapiens reached the higher latitudes of Europe by 45,000 years ago

The transition from the Middle to Upper Palaeolithic in Europe is marked by the regional disappearance of Neanderthals and the spread of *Homo sapiens*. While *H. sapiens* appeared in eastern Europe earlier, late Neanderthals persisted in western Europe for several millennia afterward. Archaeological evidence reveals diverse technocomplexes during this transition, making it challenging to link behavioral adaptations to specific hominin groups. The Lincombian-Ranisian-Jerzmanowician (LRJ) technocomplex, found in northwestern and central Europe, is one such example where the creators remain uncertain. This study focuses on the Ilsenhöhle site in Ranis, Germany, a key LRJ site, to identify the makers of the LRJ technocomplex and contribute to our understanding of the complex demographic and technological dynamics of this transitional period. Understanding the timing and nature of *H. sapiens* expansion into Europe is crucial for reconstructing the dynamics of human dispersal and interaction with Neanderthals. This study aims to address the question of who created the LRJ technocomplex by applying cutting-edge techniques of ancient DNA analysis, proteomics, and radiocarbon dating to human remains directly associated with the LRJ assemblage at Ranis.

Previous research on the Middle to Upper Paleolithic transition in Europe has highlighted the chronological overlap between *Homo sapiens* and Neanderthals, along with evidence of interbreeding. However, the precise timing and patterns of *H. sapiens* expansion remain debated, particularly regarding the spread into higher latitudes. The LRJ technocomplex has been variously attributed to either Neanderthals or *H. sapiens*, based on interpretations of its lithic technology. Some researchers suggest a Middle Paleolithic origin, while others view it as an early Upper Paleolithic development. The existing chronological data for LRJ assemblages (around 44,000–41,000 calibrated years before present (cal BP)) overlaps with the known presence of both Neanderthals and *H. sapiens* in Europe, further complicating the attribution of the LRJ. This study builds upon previous research by applying new dating methods and genetic analyses to clarify the identity of the LRJ makers and refine our understanding of the chronological overlap between the two hominin groups.

The research employed a multi-faceted approach combining archaeological excavation, radiocarbon dating, proteomic analysis, and ancient mitochondrial DNA (mtDNA) sequencing. Excavations at the Ilsenhöhle site in Ranis, Germany, focused on reopening the main 1934 trench and excavating adjacent areas to bedrock. This aimed to clarify the stratigraphy and chronology of the site and recover human remains associated with the LRJ assemblage. A total of 28 radiocarbon dates were obtained from newly excavated material, including human bones, modified bones, and charcoal. These dates were used to construct a chronological model of the site. Proteomic screening using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) was performed on bone specimens from both the 2016-2022 and 1932-1938 excavations to identify hominin remains. Eleven hominin remains were tested for ancient mtDNA preservation. High-throughput sequencing was used to reconstruct near-complete mtDNA genomes from ten of these samples. Phylogenetic analysis was conducted to determine the mtDNA haplogroups and evolutionary relationships of the Ranis hominins. Additional analyses included zooarchaeological studies, sedimentology, micromorphology, and stable isotope analysis to reconstruct the environmental context and site use patterns.

The study identified thirteen hominin bone specimens from the Ranis site, including four discovered through proteomic methods. Radiocarbon dating of these remains, along with other materials from the site, indicates that the LRJ layers (layers 9 and 8) date to 47,500–45,770 cal BP and 46,820–43,260 cal BP, respectively. Ancient mtDNA analysis confirmed that these hominins were *Homo sapiens*, demonstrating that *H. sapiens* were present at the site during the LRJ occupation. Phylogenetic analysis of the ten mtDNA genomes revealed that nine belonged to haplogroup N, clustering with the Zlatý kůň individual from the Czech Republic (around 45,000 years before present), while one belonged to haplogroup R. The faunal assemblage indicates a predominance of reindeer, along with other species consistent with central European fauna during Marine Isotope Stage 3. Stable isotope analyses of equid teeth suggest a temperature decline during the LRJ occupations, indicating a highly seasonal subarctic climate. The findings confirm that the LRJ technocomplex at Ranis was produced by *H. sapiens*, challenging the hypothesis that Neanderthals disappeared from northwestern Europe long before *H. sapiens* arrival.

The discovery of early *Homo sapiens* remains associated with the LRJ technocomplex at Ranis significantly advances our understanding of the Middle to Upper Paleolithic transition in Europe. The radiocarbon dates firmly establish the presence of *H. sapiens* in central Europe by at least 45,000 years ago, considerably earlier than previously suggested for higher latitudes. This challenges the previously held view that a chronological hiatus existed between Neanderthal and *H. sapiens* occupations in northwestern Europe. The mtDNA data, linking the Ranis *H. sapiens* to a wider population network in eastern and central Europe, supports the notion of a rapid expansion of *H. sapiens* into these regions. The zooarchaeological and environmental data suggest that the early *H. sapiens* presence at Ranis was represented by small groups with a hunting-gathering subsistence strategy adapted to a cold, seasonal climate. Further research is needed to determine the broader implications of these findings for models of *H. sapiens* dispersal and interaction with Neanderthal populations.

This study provides definitive evidence that *Homo sapiens* were present in central Europe by 45,000 years ago, associated with the LRJ technocomplex. This pushes back the known timing of *H. sapiens* expansion into higher latitudes and refutes the hypothesis of a substantial chronological gap between Neanderthal and *H. sapiens* occupations in this region. The mtDNA data suggests links to wider population networks in eastern and central Europe. Future research should focus on comparing the LRJ with other contemporary technocomplexes, such as the Szeletian and Altmühlian, to better understand the cultural and technological landscape of this critical transitional period. Further investigations into the interactions between early *H. sapiens* and Neanderthals are also warranted.

The study is limited by the relatively small sample size of human remains analyzed. While the application of multiple dating methods and genomic analyses strengthens the findings, a larger sample of hominin remains and associated artifacts would be needed for a more comprehensive understanding. The fragmented nature of some bone remains may have influenced some analyses. Furthermore, the possibility of post-depositional mixing of artifacts and human remains cannot be entirely ruled out, although the radiocarbon dates and stratigraphic integrity of the site suggest such issues were minimal.