Genetic characterization and determination of the number of individuals by molecular analysis in a prehistoric finding

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Abstract

The present study focuses on the genetic analysis of skeletal human remains exhumed from a ritual burial located in Los Cercados Chalcolithic site (3970 ± 60 BP) (Valladolid, Central Spain). In this burial different pottery and animal remains were found, configuring a complex ritual, accompanied by scarce human remains, concretely a maxilla and three skulls without maxilla.

The most striking aspect of these human remains was the different impact trauma signs on the back side of the skulls. The anthropological analysis established that the skulls were typical feminine, The bad state of preservation of the maxilla did not allowed to assign this to any of the three skulls. So, it was not possible to determine the number of individuals by anthropological methodology.

However, we could determine the number of individuals by the genetic analysis of autosomal STRs and mitochondrial DNA on the skeletal remains. It was possible to assign the maxilla to one of other three human skulls. On the other hand, we have been able to verify the sex of each individual by molecular analysis. Finally, a kinship analysis among the individuals was performed using a specific software (Familias 3.0), resulting in a possible sibling relationship between two of the individuals.

Introduction

The site of Los Cercados (Mucientes, Valladolid, Central Spain), specially its A2/As pit, is one of the most relevant Chalcolithic Pre-Bell-Beaker culture sites of the Central Plateau of the Iberian Peninsula [1].

A2/AS constitute a secondary multiple burial, where osteological remains of three human skulls and one superior maxilla were exhumed [1]. The human bones − attributed to women and with clear signs of lethal trauma − were accompanied in a pit by a highly selected set of materials, which has led to the interpretation of this case as an episode of ritualized gender violence [1].

We could assume the hypothesis that the maxilla belongs to one of the three skulls, but due to the bad preservation of the remains, it was not possible to assign the maxilla to any of the skulls through anthropological methods. So, one of the objectives of the present work was the determination of the number of buried individuals.

The second goal of this study was the molecular sex determination.

Finally, it is important to know if the three or four people of the burial shared some kinship.

Section snippets

Materials and methods

Skeletal human remains were referred as 1CER, 2CER, 3CER and 4CER. The three first remains corresponded to the three skulls and the fourth corresponded with the maxilla. We selected two bone samples (parietal and petrous bones) from each skull and two teeth from the maxilla, with the aim of replicating the results.

One sample of each individual was processed in the Forensic and Population Genetics Laboratory of the Toxicology and Health Legislation Department, Medicine School, Complutense

Results

mtDNA, consensus autosomal STR profiles (including the amplification of a fragment of amelogenin gen) and X-INDELs consensus profiles are summarized on Table 1.

Regarding to the mtDNA results, it is possible to observe that HVI mtDNA profiles were obtained for all individuals, but it was not possible to assign a HVII profile to 4CER.

If comparing the autosomal STR profiles, it is possible to see that 2CER and 4CER share almost the same profile, except a set of markers with possible allelic

Discussion

Focusing on HVI results, it is possible to appreciate that 2CER and 4CER share the same HVI mtDNA. There does not seem to exist any other relationship along the maternal line among the individuals; except for 2CER-4CER, it is discarded any other kinship along the same maternal lineage.

X-INDELS provided a general idea of the similarity of the genetic profiles, but it was not possible to perform the kinship analysis with these X-chromosome markers because it does not exist a reference database of

Conflict of interest statement

None.

Acknowledgements

This work was funded by the projects HAR2009-10105 and HAR2013-41851-P (MINECO, Spanish Government) and BES2010-035322FPI grant for Sara Palomo Díez. And by G/6401400/8000 research project (Banco Santander-Universidad Complutense de Madrid, Spain) for Cláudia Gomes.

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