William
Goodwin*, Igor V. Ovchinnikov*†, Anders
Götherström‡, Galina P. Romanova§,
Viktor M. Kharitonov||, Kerstin Lidén‡,
* Human
Identification Centre, University of Glasgow, Glasgow G12 8QQ, Scotland, UK
†
Department of Medicine, Columbia University, New York, NY 10032, USA
‡
Archaeological Research Laboratory, Stockholm University, 106 91 Stockholm, Sweden
§
Institute of Archaeology, Moscow 117036, Russia
||
Institute and Museum of Anthropology, Moscow State University, Moscow 103009, Russia
-------------------------------------
Over the past ten years the analysis of mitochondrial DNA (mtDNA) has become widespread when dealing with very low quantities of and/or highly degraded DNA. The advantages of mtDNA for this type of analysis have been well documented and include its high copy number, which improves the chance of retrieving DNA and also its highly polymorphic and therefore informative nature. For these reasons mtDNA has been widely used in the study of ancient DNA (aDNA). The isolation of DNA from palaeontological samples has made it possible to study the genetic relationship of extinct species in comparison to their extant relatives.
Isolation and Analysis of Ancient DNA
(aDNA)
The field of aDNA analysis has been controversial since its inception which followed the development of the polymerase chain reaction (PCR). The PCR reaction makes it possible to take in theory just one molecule of DNA and then to copy the molecule repeatedly until there are several million copies, which can then be analysed. The great problem with studying aDNA is that in many cases will be no endogenous (fossil) DNA to study, however, modern DNA may well contaminate any DNA extract resulting in false results, the PCR reaction will amplify the contaminating DNA as well as the fossil DNA. Stringent precautions therefore need to be followed in order to validate any result. This problem is particularly acute when studying ancient hominids as modern human DNA is pervasive both inside and outside of the laboratory (for a review of the mechanisms of DNA preservation and authenticity of aDNA ref. 1).
In 1997 aDNA was reported to have been successfully isolated from the Neanderthal recovered from the Feldhofer Cave in Germany (2). With only one Neanderthal sequence and no information about the molecular variation of Neanderthals, comparisons with modern human DNA could only be taken so far. Also, because of a number of unsubstantiated claims from the field of aDNA over the previous years, some skepticism remained about the source of the “Neanderthal DNA” despite the stringent controls employed.
A
second Neanderthal recovered during the excavation of the Mezmaiskaya Cave (3), in the
northern Caucasus in southern Russia that displayed good morphological and chemical
preservation was successfully analysed (4). This
was compared to the previously reported Neanderthal and also to modern human mtDNA. The two Neanderthal’s mtDNA were more closely
related to each other that to any modern human mtDNA.
A third successfully analysed Neanderthal (5), from Vindija Cave was also
more similar to the two Neanderthals than to modern human mtDNA. The Mezmaiskaya Neanderthal differs from the
Vindija Neanderthal by only 6 substitutions (as does the Feldhofer Neanderthal over the
same region). All three Neanderthal share
the insertion at position 16,263, relative to the CRS (Cambridge reference sequence) and
overall appear to be closely related to each other and distant to modern humans. A more rigorous phylogenetic analysis was
undertaken using the computer program PAUP. When
the Neanderthal sequences were compared to modern human sequences the Neanderthals formed
a distinct group to all modern humans (Figure 1).
The DNA recovered
from the Mezmaiskaya Neanderthal was very similar to the Feldhofer and Vindija
Neanderthal. Therefore it can be concluded
with a high degree of confidence that Neanderthal DNA has been recovered and that this is
not some kind of peculiar contamination. It
is also possible to say that the Neanderthal DNA is different from modern human mtDNA,
forming a distinct group. Based on this
evidence it is not possible to say whether Neanderthals and modern humans did interbreed,
however based on the Neanderthal and modern humans analysed to date it is possible to
conclude that Neanderthals did not pass any of there mtDNA on into the modern European
mtDNA pool.
The conditions
that maximise the chance of DNA preservation (6-7), in particular low thermal age, are
present in many sites in the Caucasian mountains. This
may facilitate the further analysis of specimens from this area in order to provide
information on the molecular diversity of the local populations of the Neanderthals.
1 Hofreiter
M. et al
Nature Reviews Genetics 2, 353-359 (2001).
2 Krings
M. et al Cell 90, 19-30 (1997).
3 Golovanova
L.V. et al Curr. Anthropol. 40, 77-86 (1999).
4 Ovchinnikov I.V. et al Nature 404, 490-493 (2000).
5 Krings
M. et al Nature Genet. 26, 144-146 (2000).
6 Collins I.S. et al Nature 410, 771-772 (2001).
7
Ovchinnikov I.V. et al Nature 410, 772
(2001).
The
Galilee Skull (the Zuttiyeh Frontal) in the Context of the Levantine Hominid Record
Yoel Rak, Sackler Faculty of Medicine, Tel Aviv University, Israel
------------------------------
The Zuttiyeh specimen known as Galilee Man was discovered in 1925 in a cave in Wadi Amud in the eastern Galilee. Not much has survived of its anatomy. The specimen consists primarily of a frontal bone, the right zygomatic bone, and sections of the sphenoid. It is only because of the paucity of the remains, which necessarily exhibit very few "taxonomic characters," and their generalized nature that we can tolerate the claim that the specimen is morphologically representative of the common ancestor of Neandertal and Homo sapiens. However, the specimen's taphonomy offers no clues as to the nature of its pelvic and cranial characteristics. In other words, did the individual possess the highly specialized pelvic architecture—such as a short pubic ramus—and the defined chin of H. sapiens or the more generalized pelvis (with a long, thin pubic ramus) and the chinless mandible of the Neandertal? Only were a chinless mandible and a long pubic ramus to accompany the Zuttiyeh specimen would we be able to designate it a common ancestor on the basis of the anatomy that we have deduced from our body of knowledge vis-à-vis the derived versus generalized characters of these two taxa.
New
excavations in the cave sediments of the ‘Kleine Feldhofer Grotte’ and
‘Feldhofer Kirche’ (Neandertal, Germany)
Ralf W. Schmitz Universität Tübingen, Institut für Ur- und Frühgeschichte und Archäologie des Mittelalters, Abteilung Ältere Urgeschichte und Quartärökologie Schloss, Burgsteige 11 D-72070 Tübingen Germany E-mail: ralf.schmitz@talknet.de
and
Landschaftsverband Rheinland,
Rheinisches Amt für Bodendenkmalpflege, Endenicher Strasse 133, D-53115 Bonn,
Germany
-------------------------------------
The Düssel stream Valley
('Neandertal' ) is situated 13 km east of Düsseldorf, Germany. In August 1856 quarrymen
discovered a human skeleton in the small cave 'Kleine Feldhofer Grotte' on the southern
bank of the Düssel. In 1864, William King assigned the name Homo neanderthalensis to the remains. Since 1991,
the Neandertal type specimen has been the subject of an interdisciplinary project of the
Rheinisches Landesmuseum Bonn, initiated and led by the author. Sections of the project
deal with, among other things, Palaeolithic cut marks on the skull and genetic analyses,
which yielded in 1997 the first DNA sequence of any Neandertal worldwide (Krings et al.
1997; 1999). Work is also being carried out to determine the age of the hitherto still
undated specimen.
The place of discovery, however, had
been regarded since the beginning of the 20th century as lost. In the autumn of 1997,
following several years of archival research, the author and his colleague J. Thissen
rediscovered the find spot of the Neandertal type specimen. During the excavations in 1997
and 2000 there appeared the cave's loamy sediment peppered with limestone rubble and
sinter, which during the limequarrying had ejected into the valley as useless material in
front of the cliff-face. A second cave-fill, which in part was interlocked with the first, originated from the adjacent cave 'Feldhofer
Kirche'. The present state of analysis indicates a settlement of the 'Feldhofer Kirche'
not only during the Upper Palaeolithic, but also during the Middle Palaeolithic. The Upper
Palaeolithic is represented by crested blades, bitruncated backed bladelets, Gravette
points, Font-Robert points, bec, pointe à face plane and fragmented points, made of bone
and ivory. This combination represents a Gravettian / Perigordian V.
The Middle Palaeolithic component
displays the following forms: bifacial scrapers, a bifacial backed knife, reworked
bifacial tools and fragments of bifacial tools, a fragment of a leaf point or a distal
fragment of a flat handaxe ('Faustkeilblatt') and circular scrapers ('Groszaki').
Moreover, up to now 60 human bone
fragments have been determined (F. H. Smith, Northern Illinois University and M. Schultz,
University of Göttingen). In 1999 J. Thissen and the author succeeded in fitting one of
the newly discovered fragments of bone onto the left femur of the Neandertal man from
1856, and during the last campaign the refitting of the left orbit has been achieved.
Among the fragments are also pieces
of a right humerus and fragments of an ulna. Since the corresponding bones of the
Neandertal man of 1856 are present, we are dealing in the case of the newly discovered
finds with the remains of a second human individual. The humerus has been dated by G.
Bonani, Swiss Federal Institute of Technology, Zurich, using the 14C-AMS method. The uncalibrated values fall around
40,000 years within the period of the Neandertals. Whether the second individual actually
is a Neandertal should be revealed by the working being carried out, including a
mtDNA-analysis.
Literature:
KRINGS, M., STONE, A., SCHMITZ, R.
W., KRAINITZKI, H., STONEKING, M. and PÄÄBO, S. 1997: Neandertal DNA Sequences and the
Origin of Modern Humans.- Cell 90: 19-30; Cambridge, Mass.
KRINGS, M., GEISERT, H., SCHMITZ, R.
W., KRAINITZKI, H. & PÄÄBO, S. 1999: DNA sequence of the mitochondrial hypervariable
region II from the Neandertal type specimen.- Proceedings of the National Academy of
Sciences, U.S.A., 96: 5581-5585;Washington.
SCHMITZ, R. W. & THISSEN, J.
2000: Neandertal. Die Geschichte geht weiter.- XX + 327 p.; 116 fig.; Heidelberg
(Spektrum).
How different were
they from us in cranial morphology? Inferring about ontogeny and phylogeny in Neandertals
vs. modern humans
--------------------------------
Cranial discrete traits (the so-called
"epigenetics") may represent a useful source of information about ontogenetic
patterns in H. sapiens and other extinct species of the genus Homo.
According to a functional approach, in fact, selected cranial discrete traits seem to
reflect different type, location, and intensity of stress placed on the cranial vault
during late foetal and early post-natal periods of bone growth. The phenomenon may be
defined as “ontogenetic stress”, since it represents
the epigenetic outcome of a contrast between different developmental pressures acting
before the complete ossification of cranial bones. Distinguishing these traits between hypostotic (weak osseous development,
retention of infantile traits) and hyperostotic (non-pathological excess of ossification)
features, variable degrees of hypostosis or "hypostotic scores" are registered
and compared in Neandertals and modern humans (both early and recent samples). Clear
distinct patterns of distribution of hypostosis in the diverse cranial regions are
observed. From these data, it appears that the architectural compromise represented by the
Neandertal cranium – that combines platicephaly with encephalisation – results
in mechanical stress. The phenomenon registered by the
occurrence of hypostosis is thus interpreted as the epigenetic outcome of a
contrast between variable pressures produced by
differences in the developmental pattern and genetic control of brain and cranial bones
respectively. In contrast, modern humans can be regarded as the result of an ontogenetic
revolution, probably involving a new regulation of cranial growth with respect to
brain development.
Manzi G. & Vienna A. 1997 - Cranial
non-metric traits as indicators of hypostosis or hyperostosis. Riv. di Antropol. 75,
41-61.
Manzi G., Gracia A. & Arsuaga J.L.
1998a - Cranial discrete traits in the Middle Pleistocene humans from Sima de los Huesos
(Sierra de Atapuerca, Spain). Does hypostosis represent any increase in "ontogenetic
stress" along the Neanderthal lineage? J. Human Evol. 38: 425-446.
Manzi G., Vienna A. & Hauser G. 1996
- Developmental stress and cranial hypostosis by epigenetic trait occurrence and
distribution: an exploratory study on the Italian Neandertals. J. Human Evol. 30: 511-527.
Sergi S. 1934 - Ossicini fontanellari
della regione del lambda nel cranio di Saccopastore e nei crani neandertaliani. Riv. di
Antropol. 30 (1933-34): 101-112.
Trinkaus E. & LeMay M. 1982 -
Occipital bunning among later Pleistocene hominids. Am. J. Phys. Anthrop. 57: 27-35.
Thick bones and thin plate splines: The
role of landmark-based and landmark-free morphometrics in the investigation of Neanderthal
morphology
Christoph P. E. Zollikofer and Marcia
S. Ponce de León
-------------------------------
Quantifying the morphology of fossil
skeletal elements requires measuring tools which adequately capture the complexity of
biological structures. Definitions of organismic geometries typically involve the concept
of landmarks, i.e. anatomical points of reference which denote homology relations between
specimens. While classical morphometry was confined to measurement of interlandmark
distances and angles, methods of new geometric morphometrics permit analysis of spatial
characteristics of entire landmark configurations, thus greatly enhancing the analytical
efficiency and sensitivity. However, landmarks only represent a small subset of the total
information contents of fossil bones. For example, morphometric properties of bony
surfaces and volumes have yet to be explored, quantified and scrutinised for homology
relations, using advanced techniques of data acquisition and visualisation. During
investigation of Neanderthal skeletal morphology, landmark-based and landmark-free methods
complement each other. Here, we report on work in progress in both areas. First, we
present a new method to quantify and visualise spatiotemporal patterns of bilateral shape
variation in fossil and extant skeletal morphologies. Second, we show how techniques of
morphometric mapping can be used to visualise patterns of bone thickness fluctuation.
Visualising patterns of shape
variation. Being
bilaterally symmetrical, structures such as the skull and mandible represent a special
case of within-specimen covariation of shape. Symmetrical left/right variation as well as
natural deviations from symmetry in the form of fluctuating or directional asymmetry
represent patterns of covariation on their own. However, in fossil specimens, diagenetic
events may have distorted the morphology post mortem,
such that deviations from symmetry do not necessarily convey a biologically relevant
signal. We therefore propose a quantitative method to render specimens symmetrical with
respect to an optimum midplane. Geometric morphometric analysis is then used to
characterise modes of symmetrical shape variation in the sample. To visualise patterns of
3-dimensional shape variation resulting from such analyses, we propose an alternative to
classical D'Arcy Thompson deformation grids. Making direct reference to the surface
structures of the organism under investigation rather than to abstract grids permits a
comprehensive visual grasp of shape change and its tentative interpretation in terms of
differential growth processes.
Morphometric mapping of bone thickness. Bone thickness is difficult to
quantify as a morphometric character, since it exhibits considerable fluctuations over
anatomical regions such as the cranial vault or the diaphyses of long bones. We propose a
method termed morphometric mapping, which permits comprehensive visualisation of
fluctuations in bone thickness and facilitates the search for locations representing
between-specimen homology.
Cranial growth in Neanderthals:
Developmental or functional constraints?
Marcia S. Ponce de León and Christoph
P. E. Zollikofer Anthropological Institute and MultiMedia Laboratory, Department of
Computer Science
University
of Zürich, CH-8057 Zürich/Switzerland
--------------------------------------
Craniomandibular growth in Neanderthals
is accompanied by considerable morphological changes, leading to formation of an array of
features which are considered to be unique for this extinct hominid taxon. Here we ask to
which extent such features are determined by taxon-specific morphogenetic programmes
and/or to which extent they reflect generalised biomechanical constraints. Specifically,
we analyse variation in bone thickness with techniques of morphometric mapping. Bone
thickness is thought to reflect functional constraints during an individual's lifetime
rather than its genetic disposition. We propose a quantitative definition of average bone
thickness and compare the development of cranial vault robusticity in an ontogenetic
series of Neanderthals and anatomically modern humans. Our findings show that bone
thickness in both species is extremely variable, suggesting highly diverse
functional/environmental constraints. Despite variability, however, vault robusticity
grows allometrically with the overall dimensions of the neurocranium, indicating basic
constraints imposed onto the design of the vault. Size- and age-corrected data of cranial
thickness further suggest that spatiotemporal patterns of bone deposition in the cranial
vault are similar in Neanderthals and AMH. These findings hint at a common ancestral
pattern of development and are in contrast to earlier statements that Neanderthals have
more robust cranial vault bones than AMH. Using clinical comparative data, we further
investigate the potential significance of patterns of local fluctuation in vault thickness
with respect to brain structure and function.
Neandertal
genetic diversity
Serre David”,
Krings Matthias”, Paunovic Maja², Capelli Cristian*, Tshentscher Frank+, Geisert
Helga**, Meyer Sonja”, von Haesler Arndt”, Grossschmidt Karl’, Possnert Göran++, Pääbo Svante”
”Max Planck
Institute of Evolutionary Anthropology, Leipzig, Germany
²Institute
of Quaternary Paleontology and Geology, Croatian Academy of Sciences and Arts, , Zagreb,
Croatia
*Institute of
Legal Medicine, Catholic University of S.Cuore, , Rome, Italy.
+Institute for
Human genetics, University Clinic, ,Essen, Germany
**Institute of
Zoology, University of Munich, , Munich, Germany
‘Institute for Histology and Embriology,
University of Vienna, Vienna, Austria
++Ångström
Laboratory, Division of Ion Physics, Uppsala, Sweden
------------------------------------------
In order to investigate the
possibility to retrieve DNA from Paleolithic human remains, we analyzed the amino acid
composition and extent of amino acid racemization in bones found at numerous localities
around the world. Samples from several locations proved to have a high content of amino
acids, an amino acid composition similar to that of the contemporary bone and a low level
of racemization of aspartic acid, alanine and leucine; all criteria compatible with a
chance of ancient DNA retrieval (Poinar et al,
1996). By contrast, at other locations, bones contained almost no detectable amino acids
making the retrieval of DNA sequences unlikely.
From some of the former sites,
mitochondrial DNA sequences have been retrieved from Neandertal bones using procedures
previously developed in our laboratory (Höss et al.,
1993, Krings et al., 1997, Hofreiter et al., 2001). For example, a bone excavated in
Vindija (Croatia), dated by AMS to more than 42000 years B.P. was used for DNA
extractions. A total of 620 bp of mitochondrial DNA sequence was determined (Krings et al, 2000). The new sequence groups with the two
other Neandertal sequences already published (Krings et
al., 1997, Krings et al., 1999, Ovchinnikov et al., 2000) using different phylogenetic
reconstruction methods. The results do not exclude that interbreeding between Neandertals
and modern humans may have taken place, but they show that even if it occurred,
Neandertals did not end up contributing mtDNA to the contemporary human gene pool.
Despite the fact that more extensive
sampling of Neandertals is obviously desirable, the current sequences indicate that: a) the diversity of Neandertal mtDNA is so
restricted that it is highly unlikely that a Neandertal mtDNA lineage was divergent enough
to form an ancestral lineage to some modern European mtDNA, and b) Neandertal may have
been similar to modern humans in having a low species-wide mtDNA diversity. In the case of
humans, the low genetic diversity seen both in mtDNA and nuclear DNA sequences is likely
to be the result of the rapid expansion from a population of small size. Thus, if the
Neandertals had a low diversity, this may indicate that they had expanded from a small
population. Analyses of further Neandertal specimens, which are in progress, will reveal
if a population history similar to that seen in modern humans underlies the reduced
diversity in Neandertals.
Höss, M. & Pääbo, S : DNA
extraction from Pleistocene bones by a silica-based purification method. Nucl. Acids Res
21, 3913-3914 (1993)
Poinar, H.N., Höss, M., Bada, J.L. & Pääbo, S.: Amino acid
racemization and the preservation of ancient DNA. Science 272: 864-866 (1996)
Krings, M., Stone, A., Schmitz, R.W.,
Krainitzki, H., Stoneking, M., and Pääbo, S.: Neandertal DNA sequences and the origin of
modern humans. Cell 90: 19-30 (1997).
Krings, M., Geisert, H., Schmitz,
R.W., Kraininzki, H., and Pääbo, S.: DNA sequence of the
Krings, M., Capelli, C.,
Tschentscher, F., Geisert, H., Meyer, S., von Haeseler, A., Grossschmidt, K., Possnert,
G., Paunovic, M. and Pääbo, S.: A view on Neandertal genetic diversity. Nature Genetics
26: 144-146 (2000).
Ovchinnikov, I.V., : Molecular
analysis of Neanderthal from the northern Caucasus. Nature 404: 490-493 (2000).
Hofreiter, M., Serre, D., Poinar, N.H.,
Kuch, M. and Pääbo, S.: Ancient DNA. Nature Genetics Review 2: 353-359 (2001).
Fragmento de hueso
frontal del Musteriense de la Cueva de Horá (Granada,
España)
Maria Haber Uriarte, Lab. Antropología Dept. CC. Morfológicas
Facultad Medicina,
----------------------------------
The Neanderthals of the Italian
peninsula: demography, morphological traits and pathology.
Francesco
Mallegni and Giandonato Tartarelli, Department of Archaeological Science,
---------------------------
All Neanderthal skeletal specimens from
Italy have been studied in this work: they have been discovered in the sites of: Caverna
delle Fate in Liguria, Buca del Tasso-Camaiore in Tuscany, Saccopastore, Grotta Guattari
and other caves of Circeo promontory in Latium, Grottoni di Calascio rock-shelter in
Abruzzo; Grotta del Poggio, Grotta Taddeo and Grotta Il Molare in Campania, Grotta Santa
Croce di Bisceglie, Grotta del Bambino, Grotta del Cavallo-Uluzzo in Puglia, Archi and
Janni di San Calogero in Calabria.
They consist of a large number of
specimens, only the French series of Neanderthal samples is larger than the Italian
collection, even if the latter is much more fragmentary. On these remains we could notice
the distinguishing features of the Neanderthal species and those describing its biological
evolution, given the long span of time to which the sites can be dated.
We also took into account
environmental stresses and pathological features through the observation of their teeth
thus recording enamel hypoplasia, pits, apical granuloma and abscesses that represent
specific markers to be taken into consideration in order to reconstruct the behavioural
pattern of Neanderthal human groups.
Microwears
analysis of Neanderthal teeth from the Italian peninsula.
Emiliano
Carnieri, Department of Archaeological
Science,University of Pisa, Italy
--------------------------------
We analysed the dental wear of some Neanderthal human samples coming from Italy (Guattari II, Guattari III, Archi, Grotta Taddeo, Grotta Cavallo) both from a macroscopical and a microscopical point of view on occlusal, buccal, lingual, mesial and distal surfaces of anterior and posterior teeth.
Microwear
patterns have been observed using SEM and stereomicroscope and several magnifications in
order to assess the kind of diet and possible extra-alimentary use of teeth. The SEM
images have been analysed and interpreted using a semiautomatic digital image reading
programme (Microwear 3.0). Our specimens show patterns and features in various percentages
(striae, pits, sulci) that could have been caused by the age of the
individuals, their geological date, or the environmental conditions in which they lived.