Molecular and cytogenetic investigations of the fragile X region including the Frax A and Frax E CGG trinucleotide repeat sequences in families multiplex for autism and related phenotypes.

We undertook molecular and cytogenetic analyses in 25 families multiplex for autism and related disorders. Three of the multiplex families exhibited fragile X, and the affected offspring all exhibited CGG triplet repeat insertion mutations in the FMR-1 gene. One of these families contained an affected pair of monozygotic female twins. Both had similar-sized CGG triplet repeat expansions, but different phenotypic manifestations. One suffered from autism and the other from mild mental retardation and marked social anxiety. PCR and Southern hybridization analysis of the CGG repeat sequences characterizing fragile X A (Frax A) and E and the methylation status of FMR-1 showed no evidence of abnormal CGG repeat expansion or FMR-1 hypermethylation in the remaining 22 multiplex families. Moreover, there was no correlation between the Frax A or E (CGG)n repeat length with affected status, nor any association with the low-level (< 3 %) expression of cytogenetic fragility at Xq27 previously reported in these families. Our findings indicate that most instances of recurrence in families multiplex for autism and related disorders are not accounted for by Frax A and E. They also indicate that the phenotypic manifestations of Frax A may be influenced by stochastic, environmental and other biological factors.