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. 2024 Dec;20(4):1276-1281.
doi: 10.1007/s12024-024-00787-7. Epub 2024 Jan 31.

Forensically relevant anatomical brain regions cannot be sub-differentiated by RNA expression analysis

Jan Euteneuer  1 , Lucas Moitinho-Silva  2 , Cornelius Courts  3   4
Affiliations

Forensically relevant anatomical brain regions cannot be sub-differentiated by RNA expression analysis

Jan Euteneuer et al. Forensic Sci Med Pathol. 2024 Dec.

Abstract

The contextualization of biological traces generated by severe head injuries can be beneficial for criminal investigations. Here we aimed to identify and validate mRNA candidates for a robust sub-differentiation of forensically and traumatologically relevant brain regions. To this purpose, massively parallel sequencing of whole transcriptomes in sample material taken from four different areas of the cerebral cortex (frontal, temporal, parietal, occipital lobe) was performed, followed by bioinformatical data analysis, classification, and biostatistical candidate selection. Candidates were evaluated by Multiplex-RT-PCR and capillary electrophoresis. Only a weak relative upregulation and solely for candidates expressed in the parietal lobe was observed. Two candidates with upregulation in the cerebellar region (PVALB and CDR2L) were chosen for further investigation; however, PVALB could not reliably and repeatedly be detected in any lobe whereas CDR2L was detectable in all lobes. Consequently, we suggest that differences in mRNA expression between four regions of the cerebral cortex are too small and less pronounced to be useful for and applicable in forensic RNA analysis. We conclude that sub-differentiation of these brain regions via RNA expression analysis is generally not feasible within a forensic scope.

Keywords: Brain regions; Forensic genetics; Head trauma; Organ tissue identification; RNA analysis; RNA sequencing.

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Conflict of interest statement

Declarations. Ethics approval: This study was performed in line with the principles of the Declaration of Helsinki. The study design and experimental procedures were approved by the ethics committees of the University Hospital of Bonn and the University Medical Center of Schleswig-Holstein. Conflict of interest: The authors declare no competing interests. Previous or partial publication of contents: Preliminary results from this study were presented at the 29th Congress of the International Society for Forensic Genetics, Washington, DC, USA, August 29th to September 2nd, 2022 (P209). Concomitantly, a non-peer-reviewed “extended abstract” titled “Towards localizing head shots – Forensic sub-differentiation of anatomical brain regions by differential RNA expression” was published in the Forensic Science International: Genetics Supplement Series ( https://doi.org/10.1016/j.fsigss.2022.10.027 ), the corresponding publication venue for the proceedings of scientific symposia.

Figures

Fig. 1
Fig. 1
Expression of PVALB. Combined depiction of electropherograms of the expression of PVALB and 18s rRNA in all brain regions (frontal lobe, occipital lobe, parietal lobe, temporal lobe) in a set of three brain samples. X-axis depicts amplicon length (80–120 bp); Y-axis depicts RFU (relative fluorescent units, between 0 and 15,000)
Fig. 2
Fig. 2
Expression of CDR2L. Electropherogram depictions of the expression of CDR2L and 18s rRNA in investigated brain regions, each combined for a set of three brain sample. X-axes depict amplicon length (100–150 bp); Y-axes depict RFU (relative fluorescent units, between 0 and 40,000). a Expression of CDR2L and 18s rRNA in frontal lobe, b expression of CDR2L and 18s rRNA in occipital lobe, c expression of CDR2L and 18s rRNA in parietal lobe, d expression of CDR2L and 18s rRNA in temporal lobe
See this image and copyright information in PMC

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