5 research outputs found
Detection of CX3CR1 single nucleotide polymorphism and expression on archived eyes with age-related macular degeneration
There is a significant genetic component in
age-related macular degeneration (AMD). CX3CR1,
which encodes the fractalkine (chemokine, CX3CL1)
receptor, has two single nucleotide polymorphisms
(SNPs): V249I and T280M. These SNPs are correlated
with other aged-related diseases such as atherosclerosis.
We have reported an association of CX3CR1 SNP and
AMD. In this study we examined CX3CR1 SNP
frequencies and protein expression on archived sections
of AMD and normal eyes. We microdissected nonretinal,
peripheral retinal and macular cells from
archived slides of eyes of AMD patients and normal
subjects. CX3CR1 SNP typing was conducted by PCR
and restriction fragment length polymorphism analysis.
CX3CR1 transcripts from retinal cells were also
measured using RT-PCR. CX3CR1 protein expression
was evaluated using avidin-biotin complex
immunohistochemistry. We successfully extracted DNA
from 32/40 AMD cases and 2/2 normal eyes. Among the
32 AMD cases, 18 had neovascular AMD and 14 had
non-neovascular AMD. The M280 allele was detected in
19/64 (32 cases x2) with a frequency of 29.7%, which
was significantly higher as compared to the frequency in
the normal population (11.2%). We detected CX3CR1
expression in the various retinal cells. CX3CR1
transcript and protein levels were diminished in the
macular lesions. This study successfully analyzed
CX3CR1 SNP and transcript expression in
microdissected cells from archived paraffin fixed slides.
Our data suggest that the M280 allele, a SNP resulting in
aberrant CX3CR1 and CX3CL1 interaction, as well as
lowered expression of macular CX3CR1, may contribute
to the development of AMD