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      High incidence of cervical ribs indicates vulnerable condition in Late Pleistocene woolly rhinoceroses

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          Abstract

          Mammals as a rule have seven cervical vertebrae, a number that remains remarkably constant. Changes of this number are associated with major congenital abnormalities (pleiotropic effects) that are, at least in humans, strongly selected against. Recently, it was found that Late Pleistocene mammoths ( Mammuthus primigenius) from the North Sea have an unusually high incidence of abnormal cervical vertebral numbers, approximately ten times higher than that of extant elephants. Abnormal numbers were due to the presence of large cervical ribs on the seventh vertebra, indicating a homeotic change from a cervical rib-less vertebra into a thoracic rib-bearing vertebra. The high incidence of cervical ribs indicates a vulnerable condition and is thought to be due to inbreeding and adverse conditions that may have impacted early pregnancies in declining populations. In this study we investigated the incidence of cervical ribs in another extinct Late Pleistocene megaherbivore from the North Sea and the Netherlands, the woolly rhinoceros ( Coelodonta antiquitatis). We show that the incidence of abnormal cervical vertebral numbers in the woolly rhinoceros is unusually high for mammals (15,6%, n = 32) and much higher than in extant Rhinoceratidae (0%, n = 56). This indicates that woolly rhinoceros lived under vulnerable conditions, just like woolly mammoths. The vulnerable condition may well have contributed to their eventual extinction.

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          Sequencing the nuclear genome of the extinct woolly mammoth.

          In 1994, two independent groups extracted DNA from several Pleistocene epoch mammoths and noted differences among individual specimens. Subsequently, DNA sequences have been published for a number of extinct species. However, such ancient DNA is often fragmented and damaged, and studies to date have typically focused on short mitochondrial sequences, never yielding more than a fraction of a per cent of any nuclear genome. Here we describe 4.17 billion bases (Gb) of sequence from several mammoth specimens, 3.3 billion (80%) of which are from the woolly mammoth (Mammuthus primigenius) genome and thus comprise an extensive set of genome-wide sequence from an extinct species. Our data support earlier reports that elephantid genomes exceed 4 Gb. The estimated divergence rate between mammoth and African elephant is half of that between human and chimpanzee. The observed number of nucleotide differences between two particular mammoths was approximately one-eighth of that between one of them and the African elephant, corresponding to a separation between the mammoths of 1.5-2.0 Myr. The estimated probability that orthologous elephant and mammoth amino acids differ is 0.002, corresponding to about one residue per protein. Differences were discovered between mammoth and African elephant in amino-acid positions that are otherwise invariant over several billion years of combined mammalian evolution. This study shows that nuclear genome sequencing of extinct species can reveal population differences not evident from the fossil record, and perhaps even discover genetic factors that affect extinction.
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            Evolution of the vertebral formulae in mammals: a perspective on developmental constraints.

            Developmental constraints refer to biases that limit phenotypic changes during evolution. To examine the contribution of developmental constraints in the evolution of vertebrate morphology, we analyzed the distribution pattern of mammalian vertebral formulae. Data on mammalian vertebral formulae were collected from the Descriptive Catalogue of the Osteological Series Contained in the Museum of the Royal College of Surgeons of England by Richard Owen (1853) and were plotted onto the most reliable mammalian phylogenetic tree based on recent molecular studies. In addition to the number of cervical vertebrae that is almost fixed to 7, we found that the number of thoracolumbar vertebrae tends to be 19 in many groups of mammals. Since fidelity of the number of thoracolumbar vertebrae was also completely maintained in Monotremata and Marsupialia, we presumed that thoracolumbar vertebral number as well as cervical vertebral number might have been fixed in the primitive mammalian lineage. On the basis of primitive vertebral formulae, we could clarify the polarity of evolution and identify several deviations from the primitive states during the mammalian evolution. The changes in the vertebral formulae in eutherian mammals seem to be lineage-specific, such that most species in Carnivora have 20 instead of 19 thoracolumbar vertebrae. Because such lineage-specific vertebral formulae contrast with the estimated distribution pattern on the assumption of evolution only through the selective pressure, we concluded that developmental constraints played an important role in the evolution of mammalian vertebral formulae. Copyright 2005 Wiley-Liss, Inc.
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              Why do almost all mammals have seven cervical vertebrae? Developmental constraints, Hox genes, and cancer.

              Mammals have seven cervical vertebrae, a number that remains remarkably constant. I propose that the lack of variation is caused by developmental constraints: to wit, changes in Hox gene expression, which lead to changes in the number of cervical vertebrae, are associated with neural problems and with an increased susceptibility to early childhood cancer and stillbirths. In vertebrates, Hox genes are involved in the development of the skeletal axis and the nervous system, among other things. In humans and mice, Hox genes have been shown also to be involved in the normal and abnormal (cancer) proliferation of cell lines; several types of cancer in young children are associated with abnormalities in Hox gene expression and congenital anomalies. In these embryonal cancers the incidence of a cervical rib (a rib on the seventh cervical vertebra, a homeotic transformation of a cervical vertebra towards a thoracic-type vertebra) appears to be increased. The minimal estimate of the selection coefficient acting against these mutations is about 12%. In birds and reptiles variations in the number of cervical vertebrae have frequently occurred and there is often intraspecific variability. A review of the veterinary literature shows that cancer rates appear lower in birds and reptiles than in mammals. The low susceptibility to cancer in these classes probably prevents the deleterious pleiotropic effect of neonatal cancer when changes in cervical vertebral number occur. In mammals there is, thus, a coupling between the development of the axial skeleton and other functions (including the proliferations of cell lines). The coupling of functions is either a conserved trait that is also present in reptiles and birds, but without apparent deleterious effects, or the coupling is new to mammals due to a change in the functioning of Hox genes. The cost of the coupling of functions in mammals appears to be an increased risk for neural problems, neonatal cancer, stillbirths, and a constraint on the variability of cervical vertebral number.
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                Author and article information

                Contributors
                Journal
                PeerJ
                PeerJ
                peerj
                peerj
                PeerJ
                PeerJ Inc. (San Francisco, USA )
                2167-8359
                29 August 2017
                2017
                : 5
                : e3684
                Affiliations
                [1 ]Naturalis Biodiversity Center , Leiden, the Netherlands
                [2 ]Department of Geology and Geoenvironment, National and Kapodistrian University of Athens , Zografou, Greece
                Article
                3684
                10.7717/peerj.3684
                5580387
                556b2a29-a2f5-4d25-a615-d7aa1febac7e
                ©2017 Van der Geer and Galis

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ) and either DOI or URL of the article must be cited.

                History
                : 4 March 2017
                : 22 July 2017
                Funding
                Funded by: Royal Museum for Central Africa Tervuren
                Award ID: BE-TAF-1649
                Funded by: Zoological Museum Copenhagen
                Award ID: DK-TAF-4886
                Funded by: National Museum for Natural History in Paris
                Award ID: FR-TAF-4104
                Funded by: Natural History Museum of Stockholm
                Award ID: SE-TAF-3009
                This work was in part supported by Synthesys travel grants to F.G. to visit Royal Museum for Central Africa Tervuren, the Zoological Museum Copenhagen, the National Museum for Natural History in Paris, and the Natural History Museum of Stockholm (Grants BE-TAF-1649, DK-TAF-4886, FR-TAF-4104, SE-TAF-3009, respectively). There was no additional external funding received for this study. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Ecology
                Evolutionary Studies
                Paleontology
                Veterinary Medicine
                Pathology

                transitional vertebrae,coelodonta,north sea,developmental constraints,neck vertebrae,extinction,neck ribs,rhinoceratidae

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