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NR 6/2004
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Analiza mutacji
genów rodopsyny i peryferyny u chorych z postacią autosomalną
dominującą zwyrodnienia barwnikowego siatkówki w rodzinach
polskich
Genetic analysis of
rhodopsin and peripherin genes in patients with autosomal
dominant retinitis pigmentosa (adRP) in Polish families
Ewa Brzeziańska1,3, Małgorzata
Zdzieszyńska2, Roman Goś2, Andrzej
Lewiński3
1Z Zakładu Tyreologii Instytutu Endokrynologii
Uniwersytetu Medycznego w Łodzi
P. o. kierownik: dr hab. n. med. Ewa Sewerynek
2Z Kliniki Okulistyki i Rehabilitacji Wzrokowej
Uniwersytetu Medycznego w Łodzi
Kierownik: prof. dr hab. n. med. Roman Goś
3Z Kliniki Endokrynologii i Terapii Izotopowej
Uniwersytetu Medycznego w Łodzi
Kierownik: prof. dr hab. n. med. Andrzej Lewiński |
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| Summary: |
Purpose: The aim of
that study was to identify the mutations in rhodopsin
and peripherin genes in Polish families with autosomal
dominant form of retinitis pigmentosa and determine the
population polimorphism in both genes in adRP families.
Material and methods: We performed ERG, visual
acuity, Goldman visual fields, intraocular pressure
measurements and fundoscopy in all the patients included
in the study. On the basis of disease history, the
families pedigree was made and the mode of inheritance
was analyzed. The molecular analysis of DNA for each
family with adRP was conducted. Genomic DNA was obtained
from leucocytes by phenol-chloroform procedure according
to Maniatis protocol. DNA was amplified by the PCR
reaction in a volume of 50 µl containing 100 ng/µl of
genomic DNA, water, Cetus buffer pH 8,4 (1n Tris, 1n
MgCl, 1n KCl, 2% gelatin), 0,25 µM of each primer, 200
µM of each of dATP, dTTP, dCTP, and dGTP and 2,5 U Taq
polymerase (Promega). For amplification of rhodopsin
gene 30 cycles of PCR were carried out. Each cycle
consisting of denaturation at 95°C for 5 min, annealing:
at 58°C (exon 1), 63°C (exon 2 i 3), 68°C (exon 4) and 2
min extension at 72°C min. For amplification of
peripherin gene 30 cycles of PCR were carried out with
annealing at 60°C. The entire PCR product was in
electrophoresis on 8% PAA. The PCR-RFLP PCR- HD PCR-SSCP
and analysis of polymorphism (CA)n dinucleotide
repetition was performed.
Results: Molecular study demonstrated, that
mutations in rhodopsin gene were cause of retinitis
pigmentosa in case of two families. In any study
families mutations in peripherin gene were not
identified. Two kinds of bases polymorphism were
identified: restriction fragments length polymorphism (RFLP)
in rhodopsin gene in exon 1 and 3 and single strand
conformation polymorphism (SSCP) in exon 1 and 3 in
rhodopsin gene and in exon 3 in peripherin gene. The
confirmed mutations in rhodopsin gene, cosegregation
with adRP, whereas two kinds of population polymorphism
did not correlate with clinical symptoms. Natural
polymorphism appeared to be a frequent feature in
rhodopsin gene while a less frequent feature in
peripherin gene.
Conclusions: Genetic investigations in patients
with adRP allow to confirm the diagnosis and evaluate
the prognosis. The mutation in rhodopsine gene should be
confirmed in directly sequencing reaction in next study. |
| Słowa kluczowe: |
gen rodopsyny, gen
peryferyny, mutacje genowe, zwyrodnienie barwnikowe
siatkówki. |
| Key words: |
rhodopsine gene,
peripherin gene, genes mutations, retinitis pigmentosa. |
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