Short Report A presenilin 1 mutation in the first case of Alzheimer’s disease: Revisited Carsten Rupp a,b , Konrad Beyreuther b , Konrad Maurer c,1 , Stefan Kins a, * ,1 a Division of Human Biology and Human Genetics, Technical University of Kaiserslautern, Kaiserslautern, Germany b Network Aging Research (NAR), Heidelberg, Germany c Department of Psychiatry, Psychosomatic Medicine and Psychotherapy, Goethe-University, Frankfurt am Main, Germany Abstract Background: Recently, a single point mutation in the presenilin 1 (PSEN1) gene of the first described Alzheimer’s disease (AD) patient Auguste D was reported by M€uller and co-workers. How- ever, the sequencing results of the DNA from a 100-year-old tissue contained some uncertainties. Methods: We heat extracted DNA from an original histological slice of Auguste D’s brain and used nested polymerase chain reaction for the amplification of different exons of genes known to be affected in familial forms of AD. Results: Our sequencing analysis did not validate the reported mutation. Furthermore, an extended sequencing analysis of Auguste D’s DNA revealed no indication of a nonsynonymous hetero- or ho- mozygous mutation in the exons of APP, PSEN1, and PSEN2 genes comprising the already known familial AD mutations. Conclusion: Despite the wealth of data from M€uller and co-workers, our results emphasize the requirement of more detailed analysis of Auguste D’s DNA in future. Ó 2014 The Alzheimer’s Association. All rights reserved. Keywords: Alzheimer’s disease; Auguste Deter; Paraffin embedded tissue 1. Introduction The neuropathological changes of Auguste D’s brain described by Alois Alzheimer have been known for a long time [1], and the cause of the early onset of the disease in the case of this first patient (51 years) has been addressed in several studies. The sequencing of Auguste D’s DNA revealed that she was homozygous for apolipoprotein E allele epsilon 3 [2]. Furthermore, the sequencing of a single exon of presenilin 2(PSEN2) from the DNA extracted from Auguste D’s brain tissue sections unveiled the wild-type sequence [3]. The same researchers reported a nucleotide substitution at posi- tion c.526 (c.526T.C) in exon 6 of PSEN1, resulting in an amino acid substitution from phenylalanine 176 to leucine (F176L) [4]. This mutation might cause the early onset of Alz- heimer’s disease (AD), but so far no genetic or biochemical evidence exists corroborating this assumption. The extraction of DNA from such old tissues is error prone and controls have only limited significance. However, the sequencing reaction of this analysis was of notable low quality. The chromatogram of the forward sequencing reaction contained, in addition to the reported mutation, two additional uncertain sites at positions c.533 and c.537. The authors interpreted these uncertainties as a consequence of compres- sion of the sequencing result. However, the observable, but unmentioned, nucleotide exchange at position c.533C.G [4] cannot be explained this way, as a compression would only account for exchanges in sequencing results from c.533 to one of the neighboring nucleotides (T and A). Notably, the mutation (c.533C.G) would generate a stop codon at position 178, resulting in a truncated presenilin 1 protein. 2. Methods 2.1. DNA extraction To extract DNA from embedded brain slices of Auguste D, the glass slide was carefully cleaned and incubated in 100% xylene. After 48 hours, the cover slip was carefully removed. After washing twice with ethanol for 10 minutes Conflict of Interest: The authors have no conflicts of interest to disclose. 1 These two authors contributed equally. *Corresponding author. Tel.: 149-631-2052106; Fax: 149-631- 2053517. E-mail address: s.kins@biologie.uni-kl.de 1552-5260/$ - see front matter Ó 2014 The Alzheimer’s Association. All rights reserved. http://dx.doi.org/10.1016/j.jalz.2014.06.005 Alzheimer’s & Dementia - (2014) 1-4