Genetic material from a baby dinosaur

A clutch of Massospondylus carinatus eggs from the Jurassic of South Africa (credit: Brett Eloff)

Recently, a lot of publicity focussed on stunning CT scans of embryos preserved in fossilised eggs of a Jurassic sauropodomorph dinosaur, which were obtained using very high energy X-rays generated by a synchrotron in France (Chapelle, K.E.J. et al. 2020. Conserved in-ovo cranial ossification sequences of extant saurians allow estimation of embryonic dinosaur developmental stages. Nature Scientific Reports, v. 10, article 4224; doi: 10.1038/s41598-020-60292-z). The images suggest that the embryos’ skulls developed in much the same way as do those of living reptiles. Within a week there emerged an even more compelling dinosaurian scoop: a fossil nestling of a duck-billed dinosaur (hadrosaur) from the Upper Cretaceous of Montana is reported to have yielded evidence for a broad spectrum of cellular materials (Bailleul, A.M. et al. 2020. Evidence of proteins, chromosomes and chemical markers of DNA in exceptionally preserved dinosaur cartilage. National Science Review, v. 7, advance publication NWZ206; DOI: 10.1093/nsr/nwz206).

Alida Bailleul, who works at the Chinese Academy of Sciences in Beijing, and fellow molecular palaeontologists from Canada, the US and Sweden, examined material from the nestling’s skull that was suspected to contain traces of cartilage. Their methods involved microscopic studies of thin sections together with staining and fluorochemical analysis of cellular material extracted by dissolving away bone tissue in acid. The same methodologies were also applied to similar material from modern emu chicks as a means of validating the results from the fossil. Staining used the same chemical that previously had revealed blood proteins in a specimen of Tyrannosaurus rex (see: Blood of the dinosaurs  in Palaeobiology, January 2011). The fluorescence approach dosed the dinosaur cartilage with antibodies against bird collagen, and revealed an immune reaction (green fluorescence) in both fossil material and that from the baby emus.

The researchers also isolated cartilage cells (chondrocytes) from the dinosaur preparations. Two stains (PI and DAPI, for short) that show up DNA were applied, giving positive responses. The PI (propidium iodide) stain is useful as it does not respond to DNA in living material, bit only to that in dead cells, thereby helping to rule out contamination with modern material. Apparently, the double-staining experiments support the presence of double-stranded material that involves at least six base pairs (of ACTG amino acids). This does not prove the existence of dinosaur DNA, but does demonstrate that the hadrosaur’s cell nuclei are preserved.

Does that suggest that the hunt is on for a dinosaur genome, with all its connotations? OK, a complete genome has been extracted from a frozen Siberian mammoth a few tens of thousand years old, which encourages ‘re-wilding’ aficionados, but that animal preserved intact cells of many kinds. A 70 Ma old dinosaur fossil, however exquisitely preserved, is mostly ‘rock’, in that preservation is through mineralisation of bone and tissue, and even cells … Moreover, it is possible that what the team found may even be material from post-mortem bacterial colonisation of any age younger than 70 Ma.

See also: De Lazaro, E. 2020. Scientists Use X-rays to Peer inside Fossilized Dinosaur Eggs Sci News, 10 April 2020; Black, R. 2020. Possible dinosaur DNA has been found. Scientific American, 17 April 2020

Further back in the Eurasian human story

About 800 to 950 thousand years (ka) ago the earliest human colonisers of northern Europe, both adults and children, left footprints and stone tools in sedimentary strata laid down by a river system that then drained central England and Wales. The fossil flora and fauna at the Happisburgh (pronounced ‘Haze-burra’) site in Norfolk suggest a climate that was somewhat warmer in summers than at present, with winter temperatures about 3°C lower than now: similar to the climate in today’s southern Norway. At that time the European landmass extended unbroken to the western UK, so any hunter-gatherers could easily follow migrating herds and take advantage of seasonal vegetation resources. These people don’t have a name because they left no body fossils. A group known from their fossils as Homo antecessor had occupied Spain, southern France and Italy in slightly earlier times (back to 1200 ka). Since the discovery of their unique mix of modern and primitive traits, they have been regarded as possible intermediaries between H. erectus and H. heidelbergensis – once supposed to be the predecessor of Neanderthals and possibly anatomically modern humans (AMH). Since the emergence about 10 years ago of ancient genomics as the prime tool in examining human ancestry the picture has been shown to be considerably more complex. Not only had AMH interbred with Neanderthals and Denisovans, those two groups were demonstrably interfertile too, and a complex web of such relationships had been pieced together by 2016. But there has been a new development.

700 ka Homo erectus from Java: a possible Eurasian ‘super-archaic’ human (credit: Gibbons 2020)

Population geneticists at the University of Utah, USA, have devised sophisticated means of making more of the detailed ATCG nucleotide sequences in ancient human DNA, despite there being very few full genomes of Neanderthals and Denisovans (Rogers, A.R. et al. 2020. Neanderthal-Denisovan ancestors interbred with a distantly related hominin. Science Advances, v. 6, article eaay5483; DOI: 10.1126/sciadv.aay5483). In Earth-logs you may already have come across the idea of the ancestral ‘ghosts’ that are represented by unusual sections of genomes from living West African people. Those sections seem likely to have resulted from interbreeding with an unknown archaic population – i.e. neither Neanderthal nor Denisovan. It now seems that both Neanderthal and Denisovan genomes also show traces of such introgression with ‘ghost’ populations during much earlier times. The ancestors of both these groups separated from the lineage that led to AMH perhaps 750 ka ago. Rogers et al. refer to the earliest as ‘neandersovans’ and consider that they split into the two groups after they entered Eurasia, at some time before 600 ka – perhaps around 740 ka. This division may well have occurred as a result of a population of ‘neandersovans’ having spread over the vastness of Eurasia and growing genetic isolation. The reanalysis of both sets of genomes show evidence of a ‘neandersovan’ population crash before the split. Thereafter, the early Neanderthal population may have risen to around 16 thousand then slowly declined to ~3400 individuals.

A ‘state-of-play’ view of human interbreeding in Eurasia since 2 Ma ago (credit: Gibbons 2020)

However, the ‘neandersovans’ did not enter a new continent devoid of hominins, for as long ago as 1.9 Ma archaic H. erectus had arrived from Africa.  Both Neanderthal and Denisovan genomes record the presence of sections of ‘super-archaic’ DNA, which reflect early  interbreeding with earlier Eurasian populations. Indeed, Denisovans seem to have repeated their ancestors’ sexual exploits, once they became a genetically distinct group.  From the ‘ghost’ DNA fragments Rogers et al. conclude that the ‘super-archaics’ separated from other humans about two million years ago. They were descended from the first ‘Out-of-Africa’ wave of humans, represented by the fossils humans from Dmanisi in Georgia (see First out of Africa, November 2003 and An iconic early human skull,  October 2013 in Earth-logs Human evolution and migrations). A measure of the potential of novel means of analysing available ancient human DNA is the authors’ ability even to estimate the approximate population size of the interbreeding ‘super-archaic’ group at 20 to 50 thousand. Long thought to be impossible, it now seems possible to penetrate back to the very earliest human genetics, and the more DNA that can be teased out of other Neanderthal and Denisovan fossils the more we will know of our origins.

See also: Gibbons, A. 2020. Strange bedfellows for human ancestors. Science, v. 367, p. 838–839; doi:10.1126/science.367.6480.838

Neanderthal Mum meets Denisovan Dad

Two bone fragments from the Denisova Cave – the former abode of an 18th century Russian hermit called Denis – in the Altai region of Siberia yielded ancient  DNA. One matches that from previously analysed Neanderthal remains and the other a genome that could only be ascribed to a hitherto unknown ancient-human population, now known as the Denisovans. Since their discovery further analysis of both modern and ancient DNA has shown that modern humans living outside of Africa contain a few percent of DNA from both ancient-human groups. Soon after leaving Africa some of their ancestors interbred with both; indeed a 40 ka-old modern-human jaw from Romania revealed genetic evidence that the individual had a Neanderthal great-great grandparent. Their descendants spread far and wide to populate Eurasia, Australasia and the Americas. Using the ancient DNA to peer back in time suggests that Neanderthals and Denisovans diverged from a common ancestor between 470 and 380 ka, itself having split from modern-human ancestry between 770 to 550 ka. Denisovan DNA also contains evidence that its ancestry included segments that could only have come from a totally unknown hominin species. Interestingly, DNA from the Neanderthal bone fragment found at Denisova contains fragments from an anatomically modern-human.

Tourists at the entrance to Denisova Cave, Rus...
Tourists at the entrance to Denisova Cave, Russia (credit: Wikipedia)

With such riches from tiny fragments of human bones unearthed from the Denisova Cave, it is no surprise that the team led by Svante Pääbo at the Max Planck Institute for Evolutionary Anthropology in Leipzig, Germany, has subsequently analysed others that showed signs of human proteins. The latest ‘takes the biscuit’. A fragment of limb bone from someone who was at least 13 years old yielded DNA commensurate with their having been the child of a Neanderthal mother and a Denisovan father (Slon, V. and 18 others 2018. The genome of the offspring of a Neanderthal mother and a Denisovan father. Nature, v. 560, published on-line; doi: 10.1038/s41586-018-0455-x). Their child was a girl, who has been nicknamed ‘Denny’ by the team, though ‘Denise’ might seem more appropriate. The only clues to what her father, or any Denisovan, might have looked like stem from a few teeth and a skull fragment from the cave that have yielded Denisovan DNA. The teeth are much larger and the skull fragment is thicker than those of Neanderthals, suggesting that Denisovans were distinctly bigger and more robust than even the sturdy Neanderthals.

The father came from a population related to a later Denisovan found in the cave – the first to be sequenced. This suggests long-term occupancy of the area by Denisovans. But his genome also carries traces of Neanderthal ancestry. Surprisingly, the mother is more closely related to Croatian Neanderthals, rather than to an earlier Neanderthal found in the cave. Neanderthals were clearly capable of migrating between Europe and eastern Eurasia; more than 5000 km in this case. Even though very few archaic humans have been genetically sequenced it is beginning to look as if genetic mixing between diverse hominin groups in the last half million years was common, when they actually met. A custom of marrying outside a closely related group (exogamy) has been popular throughout recorded history; indeed it makes sound genetic sense. With the tiny human population density during the Late Pleistocene, it may then have been cause for mutual celebration.  As documented in Chapters 2 and 3 of David Reich’s Who We Are and How We Got Here (Oxford University Press, 2018) human origins since about 470 ka until the present chart a history of episodic migrations and genetic mixing that certainly makes nonsense of earlier ideas of ‘racial purity’ and casts doubt even on the term ‘species’ as regards members of the genus Homo.

If we are ever to discover who the Denisovans were and what they looked like, the evidence is likely to come from East Asia at latitudes where climate favours preservation of DNA. Advanced sequencing equipment and techniques are now operational in China, where suspected Denisovan remains have been found

See also: Warren, M. 2018. First ancient-human hybrid. Nature, v. 560, p. 417-418; doi: 10.1038/d41586-018-06004-0); Sample, I. 2018. Offspring of Neanderthal and Denisovan identified for first time. The Guardian (22 August 2918).

A revised and updated edition of Steve Drury’s book Stepping Stones: The Making of Our Home World can now be downloaded as a free eBook

Denisovan(?) remains in the garden

On the edge of the small town of Lingjing near Xuchang City in Henan Province, China, local people have long practiced intensive vegetable gardening because the local soil is naturally irrigated by the water table beneath the flood plain deposits of the Yinghe River. In the mid 1960s, around a small spring, they began to find dozens of small stone tools together with animal bones. Only in 2005, after the spring had stopped flowing, did systematic excavation begin (Li, Z.-Y. et al. 2017. Late Pleistocene archaic human crania from Xuchang, China. Science, v. 355, p. 969-972; doi: 10.1126/science.aal2482) About 3.5 m below the surface tools and bone fragments, including one with a carved representation of a bird, occurred just above the base of the modern soil profile. Radiocarbon dating of charcoal from the layer clustered around 13 500 years ago, just before the start of the Younger Dryas cooling episode; probably products of modern humans, although no human remains were found in the layer. Continued excavation penetrated sediments free of fossils and tools down to a depth of 8 m, when stone tools and bone fragments began to turn up again through the lowest 2 m of sediment. Optically stimulated luminescence (OSL) dating of mineral grains, which shows the last time that sediments were exposed to sunlight, produced much older dates between 78 to 123 ka. The thousands of stone flakes and cores, and cut marks on the animal bones found through the fossil-rich layer suggests that this was a site long used for tool making and food preparation, that had begun in the last interglacial period. Among the bones were fragments of the crania of as many as five individual humans.

Who were they? Their age range is tens of thousands of years before anatomically modern humans began to migrate into east Asia, so they are likely to have been an earlier human group. Homo erectus is known to have inhabited China since as early as 1.6 Ma ago and may be a possibility. The other possible group are the Denisovans, known only from their DNA in a small finger bone from a cave in eastern Siberia. Fragments of Denisovan DNA are famously present in that of many living indigenous people from eastern Asia, Melanesia and the Americas, but hardly at all in west Asians and Europeans. They also interbred with Neanderthals and may share a common ancestor with us and them, who lived about 700 ka ago.

Map showing the proportion of the genome inferred to be Denisovan in ancestry in diverse non-Africans. The color scale is not linear to allow saturation of the high Denisova proportions in Oceania (bright red) and better visualization of the peak of Denisova proportion in South Asia. (Credit: Sankararaman et al./Current Biology 2016;  http://dx.doi.org/10.1016/j.cub.2016.03.037)
Map showing the proportion of the genome inferred to be Denisovan in ancestry in non-Africans. The color scale ranges from black – 0, through greens – present to red – highest . (Credit: Sankararaman et al./Current Biology 2016; http://dx.doi.org/10.1016/j.cub.2016.03.037)

Unfortunately the human bones are completely fragmented and lack any teeth, jaw bones or elements of the face. However, the Chinese-US team used sophisticated computer refitting of CT-scanned fragments to reconstruct two of the crania, revealing one individual with prominent brow ridges and a flat-topped skull extended towards the back, similar to that of Neanderthals but with a much larger brain than H. erectus. The semi-circular canals associated with the ears, but used in balancing, are well preserved and also resemble those of Neanderthals. Yet east Asia has yielded not a single Neanderthal fossil. Could these be the elusive Denisovans? Even if more diagnostic bones turn up, especially teeth, such is the state of late hominin taxonomy that only DNA will provide definitive results: the Denisovans are defined entirely by DNA. The authors, perhaps wisely, do not speculate, but others may not be able to resist the temptation.

For more information on recent human evolution see here.

Gibbons, A. 2017. Close relative of Neandertals unearthed in China. Science, v. 355, p. 899; doi: 10.1126/science.355.6328.899