MeCP2 mutant protein is expressed in astrocytes as well as in neurons and localizes in the nucleus.

The MECP2 gene, located at Xq28, encodes methyl-CpG-binding protein 2 (MeCP2), which is frequently mutated (up to 90%) in Rett syndrome (RTT). RTT is a progressive neurodevelopmental disorder, which affects primarily girls during early childhood and it is one of the most common causes of mental retardation in females. R270X is one of the most frequent recurrent MECP2 mutations among RTT cohorts. The R270X mutation resides within the TRD-NLS (Transcription Repression Domain-Nuclear Localization Signal) region of MeCP2 and causes a more severe clinical phenotype with increased mortality as compared to other mutations. To evaluate the functional role of the R270X mutation, we generated a transgenic mouse model expressing MeCP2(270_EGFP) (human mutation equivalent) by BAC recombineering. The expression pattern of MeCP2(270_EGFP) was similar to that of endogenous MeCP2. Strikingly, MeCP2(270_EGFP) localizes in the nucleus, contrary to the conjecture that R270X could cause disruption of the NLS. In primary hippocampal cells, we show that MeCP2(270_EGFP) was expressed in astrocytes by colocalization with the astrocyte-specific marker glial fibrillary acidic protein. Our data showing expression of MeCP2(270_EGFP) in transgenic mice astrocytes further reinforce the recent findings concerning the expression of MeCP2 in the glial cells.

[Clinical aspects and genetics of Williams-Beuren syndrome. Clinical and molecular genetic study of 44 patients with suspected Williams-Beuren syndrome]. / Klinik und Genetik des Williams-Beuren-Syndroms. Klinische und molekulargenetische Untersuchung von 44 Patienten mit Verdacht auf WBS.

BACKGROUND: The suspected diagnosis of Williams-Beuren syndrome (WBS), which is a retardation syndrome with great clinical variability, was cause for comparison of molecular genetic, molecular cytogenetic analysis to clinical symptoms. The results of the genetical analysis of a microdeletion of the elastin gene region on chromosome 7 were compared to the clinical symptoms. Are there any differences between symptoms in case of deletion or non-deletion? How informative are the molecular genetic, molecular cytogenetic analysis? PATIENTS AND METHODS: 44 patients with suspected diagnosis of WBS were examined using molecular genetic and molecular cytogenetic methods. The clinical symptoms as general symptoms, heart anomaly, dysmorphic signs and unusual neurobehavioural features were reported during clinical investigation in standardized questionnaires. The genomic DNA of the patients and their parents was analyzed using microsatellite markers. In some cases (e.g. uninformative microsatellite studies) we also used fluorescence in situ hybridization (FISH) with an elastin gene probe and performed a conventional chromosome banding analysis. RESULTS: 15 patients had a microdeletion. 4 patients had a deletion of the paternal allel and 7 patients showed the deletion of the maternal allel. The polymorphisms were of limited informativeness. In 2 cases microsatellite analysis was not able to determine whether the paternal or the maternal allel had been lost. In 2 cases the microsatellite analysis was uninformative so that FISH analysis was performed. All FISH analysis performed had an informative result. 80% of the children with a microdeletion of chromosome 7q11.23 showed the typical dysmorphic signs, 70% exhibited the typical WBS behaviour pattern, 50% had a specific heart anomaly. In contrast, in the group of children without a chromosomal microdeletion only 30-40% showed typical dysmorphic signs, only 10% had a typical heart anomaly and none of them showed specific behavioural changes. We found no indication to association of specific symptoms with paternal versus maternal origin of the deletion. The FISH analysis combined with a conventional chromosome banding analysis is very informative for diagnostic values. The results are compared to data of literature. CONCLUSIONS: Children with developmental retardation and WBS dysmorphic signs and an unusual behaviour should be examined by a molecular cytogenetic FISH analysis. If a microdeletion of band 7q11.23 is found a special cardiologic examination should be offered.

SCA6 is caused by moderate CAG expansion in the alpha1A-voltage-dependent calcium channel gene.

Recently, moderate (CAG)>20 repeat expansions in the alpha1A-voltage-dependent calcium channel gene (CACNL1A4) have been identified in a previously unmapped type of SCA which has been named SCA6. We investigated the (CAG)n repeat length of the CACNL1A4 gene in 733 patients with sporadic ataxia and in 46 German families with dominantly inherited SCA which do not harbor the SCA1, SCA2, or MJD1/SCA3 mutation, respectively. The SCA6 (CAG)n expansion was identified in 32 patients most frequently with late manifestation of the disease. The (CAG)n stretch of the affected allele varied between 22 and 28 trinucleotide units and is therefore the shortest trinucleotide repeat expansion causing spinocerebellar ataxia. The (CAG)n repeat length is inversely correlated with the age at onset. In 11 parental transmissions of the expanded allele no repeat instability has been observed. Repeat instability was also not found for the normal allele investigating 431 meioses in the CEPH families. Analyzing 248 apparently healthy octogenerians revealed one allele of 18 repeats which is the longest normal CAG repeat in the CACNL1A4 gene reported. The SCA6 mutation causes the disease in approximately 10% of autosomal dominant SCA in Germany. Most importantly, the trinucleotide expansion was observed in four ataxia patients without obvious family history of the disease which necessitates a search for the SCA6 (CAG)n expansion even in sporadic patients.

SCA2 trinucleotide expansion in German SCA patients.

Autosomal dominant spinocerebellar ataxias (SCA) are a group of clinically and genetically heterogeneous neurodegenerative disorders which lead to progressive cerebellar ataxia. A gene responsible for SCA type 2 has been mapped to human chromosome 12 and the disease causing mutation has been identified as an unstable and expanded (CAG)n trinucleotide repeat. We investigated the (CAG)n repeat length of the SCA2 gene in 842 patients with sporadic ataxia and in 96 German families with dominantly inherited SCA which do not harbor the SCA1 or MJD1/SCA3 mutation, respectively. The SCA2 (CAG)n expansion was identified in 71 patients from 54 families. The (CAG)n stretch of the affected allele varied between 36 and 64 trinucleotide units. Significant repeat expansions occurred most commonly during paternal transmission. Analysis of the (CAG)n repeat lengths with the age of onset in 41 patients revealed an inverse correlation. Two hundred and forty-one apparently healthy octogenerians carried alleles between 16 and 31 repeats. One 50-year old, healthy individual had 34 repeats; she had transmitted an expanded allele to her child. The small difference between 'normal' and disease alleles makes it necessary to define the extreme values of their ranges. With one exception, the trinucleotide expansion was not observed in 842 ataxia patients without a family history of the disease. The SCA2 mutation causes the disease in nearly 14% of autosomal dominant SCA in Germany.