Generation of a set of induced pluripotent stem cell lines from two Alzheimer disease patients carrying APOE4 (MLUi007-J; MLUi008-A) and healthy old donors carrying APOE3 (MLUi009-A; MLUi010-B) to study APOE in aging and disease.

Late-onset Alzheimer disease (LOAD) is the most frequent neurodegenerative disease, and the APOE ε4 allele is the most prominent risk factor for LOAD. Four human induced pluripotent stem cell (iPSC) lines MLUi007-J, MLUi008-B, MLUi009-A, and MLUi010-B were generated from LOAD patients and healthy matched donors by reprogramming of B-lymphoblastoid cells (B-LCLs) with episomal plasmids. The application of B-LCLs holds a great promise to model LOAD and other diseases because they can easily be generated from primary peripheral blood mononuclear cells (PBMCs) by infection with the Epstein-Barr virus (EBV).

Accumulation of glycated proteins suggesting premature ageing in lamin B receptor deficient mice.

Accumulation of advanced glycation end products (AGEs) is accompanied by increased free radical activity which contributes to ageing and the development or worsening of degenerative diseases. Apart from other physiological factors, AGEs are also an important biomarker for premature ageing. Here we report protein modifications (glycation) in a mouse model of lamin B receptor deficient ic J /ic J mice displaying skin defects similar to those of classical progeria. Therefore, we analysed AGE-modifications in protein extracts from various tissues of ic J /ic J mice. Our results demonstrated that pentosidine as well as argpyrimidine were increased in ic J /ic J mice indicating a modification specific increase in biomarkers of ageing, especially derived from glycolysis dependent methylglyoxal. Furthermore, the expression of AGE-preventing enzymes (Glo1, Fn3k) differed between ic J /ic J and control mice. The results indicate that not only lamin A but also the lamin B receptor may be involved in ageing processes.

Molecular cytogenetic characterization of a familial pericentric inversion 3 associated with short stature.

Short stature refers to the height of an individual which is below expected. The causes are heterogenous and influenced by several genetic and environmental factors. Chromosomal abnormalities are a major cause of diseases and cytogenetic mapping is one of the powerful tools for the identification of novel disease genes. Here we report a three generation family with a heterozygous pericentric inversion of 46, XX, inv(3) (p24.1q26.1) associated with Short stature. Positional cloning strategy was used to physically map the breakpoint regions by Fluorescence in situ hybridization (FISH). Fine mapping was performed with Bacterial Artificial Chromosome (BAC) clones spanning the breakpoint regions. In order to further characterize the breakpoint regions extensive molecular mapping was carried out with the breakpoint spanning BACs which narrowed down the breakpoint region to 2.9 kb and 5.3 kb regions on p and q arm respectively. Although these breakpoints did not disrupt any validated genes, we had identified a novel putative gene in the vicinity of 3q26.1 breakpoint region by in silico analysis. Trying to find the presence of any transcripts of this putative gene we analyzed human total RNA by RT-PCR and identified transcripts containing three new exons confirming the existence of a so far unknown gene close to the 3q breakpoint.

Paramutation-like effects at the mouse scapinin (Phactr3) locus.

Paramutation-like phenomena have been extensively studied in plants and so far described for a very few engineered loci in the mouse. Here we report an allele-specific expression analysis of the Phactr3 (phosphatase and actin regulator 3) locus by identifying the first internal mouse transcripts with a paramutation-like effect not associated with transgenic or knockout mice. In our previous work, we showed that the Phactr3 gene was mainly transcribed in the brain, exhibiting a complex genomic organisation with four alternatively spliced leader exons. Due to the location of the Phactr3 gene in the distal imprinting region on mouse chromosome 2, we generated a mouse model to investigate the possible parental influence on the allelic expression pattern by reciprocally mating NMRI mice and Mus musculus castaneus. We were able to identify a single-nucleotide polymorphism in leader exon 1C representing restriction fragment length polymorphism. After reverse transcription PCR, NMRI and M. musculus castaneus showed a homozygous restriction pattern according to their genotype. Unlike this, reverse transcription PCR products of the F1 hybrids of both crosses were transcribed from the NMRI allele only. Therefore, the Phactr3 exon 1C splice variant is potentially strain specific regulated, leading to the expression of only one allele of the reciprocal crosses. So far, this has not yet been described for an internal mouse gene. These results potentially provide new insight into non-Mendelian inheritance in mammals and may serve also as a model for investigating the regulation of allele-specific expression.