Mutations in Norrin signaling genes (
NDP,
FZD4 and
LRP5) have been found in patients with familial exudative vitreoretinopathy (FEVR) and the altered signaling is suspected to play
a critical role in its pathogenesis. To better understand this relationship, we systematically performed functional analyses
on previously identified single nucleotide variants of
LRP5,
FZD4 and
NDP, utilizing the Norrin dependent Topflash reporter assay. Cell surface binding assays and protein electrophoresis analysis
of Norrin were also performed. Seven causative mutations and five possibly causative but indecisive variants were examined.
We found: (1) a nonsense mutation in
FZD4 completely abolished its signaling activity, while single missense mutations in
LRP5 and
FZD4 caused a moderate level of reduction (ranging from 26 to 48, 36% on average) and a double missense mutation in both genes
caused a severe reduction in activity (71%). These observations correlated roughly with clinical phenotypes. (2) A mutational
effect is suggested in four of five indecisive variants by signaling reductions comparable to those of missense mutations.
(3) Norrin mutants demonstrated variable effects on signal transduction, and no apparent correlation with clinical phenotypes
was observed. (4) The Norrin mutants examined demonstrated impaired cell surface binding, and some may have partially lost
their ability to form a complex with unknown high molecular weight material(s). Our results illustrate the nature of FEVR
in relation to Norrin signaling and further suggest the complexity of its disease causing mechanism.