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

Dinosaurs in the flesh and feathers

Until only a few decades ago artistic portrayals of dinosaurs had them as leathery and scaled like lizards or crocodiles, as indeed rare examples of their fossilized skin seemed to suggest. The animatronic and CGI dinosaurs of the first Jurassic Park film were scary, but brownish grey. Later films in the franchise had them mottled and sometimes in colour, but still as mainly scaled leathery monsters. Reality soon overtook imagination as more and more exquisitely preserved fossils of small species were turned up, mainly in China, that were distinctly furry, fuzzy or feathered as shown below in a Microraptor gui fossil. It is now well-established that birds arose in the Jurassic from saurischian  dinosaurs, the order that also included all of the large carnivorous dinosaurs as well as the many more nimble and diminutive ones whose feathers sometimes conferred an ability to glide or fly. Even the other main order, the ornithischia noted for hugeness and herbivory, has yielded fossil skin that suggest furry or feathered pelts. Once fur and feathers had been found, the next big issue became whether or not dinosaurs may have been as gaudy as many modern birds.

 

Fossil of a feathered dinosaur Microraptor gui from the early Cretaceous Jiufotang Formation in China (source: Wikipedia)
Fossil of a feathered dinosaur Microraptor gui from the early Cretaceous Jiufotang Formation in China (source: Wikipedia)

One of the first palaeobiologists to become immersed in the search for colourful dinosaurs was Jakob Vinther, now of Britain’s Bristol University. In The March 2017 issue of Scientific American he summarises the progress that he and his colleagues have made (Vinther, J. 2017. The true colors of dinosaurs. Scientific American, v. 316(3), p. 42-49). On his account, the major breakthrough was Vinther’s discovery of tiny spherules in fossilised octopus ink that were identical to the granules of the pigment melanin that give the famous cephalopod ‘smoke screen’ its brownie-black colour. Melanin, or more precisely the melanosomes in which it is enclosed, is a key to coloration throughout much of the animal kingdom, especially in fur and feathers. There are two basic kinds, one conferring blackness and the other that imparts a rusty red hue, which combined with paleness due to lack of melanin together produce a gamut of greys, reds, browns oranges and yellows.  Elongated melanosomes when lined up produce the phenomenon of interference fringes that yield iridescence, responsible for the bright colours of starlings, hummingbirds and some ducks when in bright light. There are other pigments, such as carotenoids (bright reds and yellows) and porphyrins (green, red and blue) that add to the gamut possible in animals, but it was melanosomes that captured Vinther’s attention because of their importance in living feather colours.

Melanosomes occur in distinctively grouped assemblages, according to actual colour, and very similar microscopic structures turned up in the first fossil bird feathers that he studied. Others had assumed that they were bacterial colonies, which had grown during decay. The breakthrough was finding a fossil bird feather in which different structures were arranged in stripes; clear signs of patterning. Vinther’s concept bears fruit in a range of furry and feathered dinosaurs. One (Anchiornis) with a black and white body and limb speckles had a bright red crest and another (Sinosauropterix) was ginger over its back with a tiger striped tail and a white underside; an example of countershaded camouflage. His team has even been able to assign different kinds of patterning to a variety of possible habitats. Given superbly preserved specimens it seems likely that dinosaur and bird coloration may be traceable back more than 200 Ma.

English: Illustration of the small theropod di...
Artist’s impression of the small theropod dinosaur Microraptor showing colours predicted by analysis of melanosomes on its feathers.(credit: Wikipedia)

Another aspect of the filmic licence of Jurassic Park was its hinging on preservation of genetic material from the Mesozoic, specifically in a parasite preserved in amber, so that the creatures could be resurrected by bio-engineering. The only relevant find is a 46 Ma old mosquito whose abdomen was blood-engorged when it was fossilised. But all that remains are high iron concentrations the organic molecule porphyrin; break-down products of haemoglobin. Given that fossil DNA can only be reassembled from millions of fragmentary strands found in fossils in digital form that corresponds to the order of AGCT nucleobases that is barely likely to be possible – the oldest full genome yet analysed is that of a 700 ka horse. However, another biological material that varies hugely among living animals, protein, has proved to be tractable, albeit in a very limited way. Frozen mammoth meat, somewhat bloody, is sometimes unearthed from Siberian permafrost, but according to one Russian mammoth expert even the best preserved is inedible.

Beyond the Pleistocene the search for fossilised proteins has been hesitant and deeply controversial, particularly in the case of that from dinosaurs, for the obvious reason of publicity suspicions. But again, it is a story of persistence and patience. Mary Schweitzer of North Carolina State University claimed in 2007 that she had found some, but was howled down by other palaeontologists on the issues of its unlikely survivability and contamination. But other researchers had pushed back the age limits. By repeating their earlier analyses with the greatest possible care Schweitzer’s team confirmed their earlier results with several strands of the protein collagen about 15 amino acids in length from an 80 Ma old duck-billed dinosaur. Moreover they were able to show a closer affinity of the partial proteins to those of modern birds than to other reptiles, tallying with tangible fossil evidence (Schroeter, E.R  and 8 others 2017. Expansion for the Brachylophosaurus canadensis Collagen I Sequence and Additional Evidence of the Preservation of Cretaceous Protein. Journal of Proteome Research, v. 16, p. 920-932). The work continues for other dinosaurs and early fossil birds, with better reason for confidence and a chance of tying-down genetic relatedness. Another approach shows that collagen may still be preserved in a Jurassic (195 Ma) sauropod dinosaur’s rib (Lee, Y-C. and 9 others 2017. Evidence of preserved collagen in an Early Jurassic sauropodomorph dinosaur revealed by synchrotron FTIR microspectroscopy. Nature Communications, v. 8 doi:10.1038/ncomms14220).

See also: Service, R.F. 2017. Researchers close in on ancient dinosaur remains. Science (News in depth), v. 355, p. 441- 442.

From small beginnings

Camarasaurus, Brachiosaurus, Giraffatitan, Euh...
Some really cool sauropods. Image via Wikipedia

The great vegetarian sauropod dinosaurs, such as Brachiosaurus, were the biggest animals to walk the Earth, weighing up to 100 tonnes, as long as 60 m from snout to the end of their tails and more than 10 m tall. So big, indeed, that even the largest contemporary predators would have been unable to get sufficient purchase with their jaws to do them much damage. This vast bulk, unlike even bigger modern whales, was unsupported by water and would have posed major problems had the sauropods not evolved very porous, low-density neck and tail bones and kept their heads small relative to the rest of their bodies. Such small heads needed to take in up to a tonne of vegetation each day to keep the monsters alive and  ambling. Their teeth are not those of a chewer, being peg- or spoon-like and pointed forwards; specialised for raking in leaves and twigs, swallowed unchewed in great gulps. Once that style of eating developed in their precursors, with no need for massive chewing muscles it became possible to evolve necks up to 15 m long with increasingly diminutive heads. Studies of large numbers of some species of sauropod precursors indicate that juveniles grew astonishingly quickly, essential if their initial vulnerability was to be outpaced; newly hatched they would have weighed little more than 10 kg. At the growth rates of modern reptiles, the largest sauropods would only have reached full size in about a century. The estimated growth rates suggest warm bloodedness, research suggesting that they maintained body temperatures up to 12°C higher than do alligators. Clearly, sauropod dinosaurs were highly specialised, and their evolution is now known to have been lengthy.

A major news feature in Nature (Heeren, F. 201. Rise of the titans. Nature, v. 475, p. 159-161) traces that evolution through several surprising stages. The earliest likely ancestors, which appear in the Late Triassic (~230 Ma), were about the size of a turkey and had teeth adapted for shredding fibrous plant material; other early dinosaurs show clear signs of a predatory lifestyle. There is a limit to the size of predators bound up with the energy balance between flesh consumption and the energy expended in casing down prey and killing them. The limits on the size of plant eaters are mechanical: how much they can stuff in and the strength of their bodies, especially legs. In a world dominated in numbers by predatory dinosaurs, the selection pressure for herbivores to outgrow them and become too big to bite would have been substantial.

Little Triassic Panphagia (‘eater of everything’) was also bipedal, but the fossil record of sauropod precursors clearly shows their growth to the order of 10 m by the Early Jurassic, but not yet a four-legged gait though they had evolved relatively short but sturdy legs, signs of mass-saving porous neck and tail bones, and jaws with a large gape suited to gulping rather than chewing. By the mid-Jurassic Period sauropods were big, strong and four-legged, and by the Cretaceous they reached unmatched dimensions with the titanosaurs. This evolutionary path was not the only one adopted for dinosaurian herbivory. The famous Iguanodon discovered in 1822 by Gideon Mantell in the Early Cretaceous of Sussex was a member of a bipedal group of herbivores, including the duck-billed dinosaurs, that spanned more or less the same time range as sauropods. Fredric Heeren’s article is accompanied by an on-line ‘tour’ of sauropod evolution (go.nature.com/c7zlct), while the American Museum of Natural History has a website for a major exhibition of sauropods (www.amnh.org/exhibitions/wld/ and http://www.youtube.com/AMNHorg ) that includes footage of  a full-scale animatronic Mamenchisaurus from China which breathes and moves, (Switek, B. 2011. Living it large: review of The World’s Largest Dinosaurs exhibition. Nature, v. 475, p. 172).

Wide-eyed dinosaurs

Dinosaur Exhibition Beijing
Image by Ivan Walsh via Flickr

One of the surprises concerning the dinosaurs was that some species were able to live at near-polar latitudes. The surprise is not about their ability to survive a cold climate for the Cretaceous world was one characterised by greenhouse conditions and ice-free polar regions swathed in forests. On top of that, evidence is accumulating that some dinosaurs at least were able to regulate their body temperature; they may have been warm-blooded. The oddity is that they were able to survive the winter darkness of latitudes above those of the Arctic and Antarctic Circles. It now seems that some groups of dinosaurs evolved excellent night-time vision (Schmitz, L. & Motani, R. 2011. Nocturnality in dinosaurs inferred from scleral ring and orbit morphology. Science, v. 332, p. 705-708). Not only did some have large eyes, but preservation of the fibrous outer ring of the eye or sclera – the ‘whites’ in our case – in some large-eyed dinosaurs shows a reduction in width that is characteristic of good scotopic or night vision. Since much of the polar ‘night’ is more like twilight than perpetually full darkness, enhanced night vision would have allowed high-latitude dinosaurs to survive winter by crepuscular feeding habits. This more or less extinguishes the notional day-night duality of terrestrial vertebrate life during the Mesozoic; dinosaurs by day and early mammals by night that allowed mammalian ancestors to escape the clutches of dinosaur predators. Indeed many Mesozoic mammals show signs of diurnality.