Abstract:
BACKGROUND:The influences of genetic and environmental factors form a clinical picture of type 2 diabetes mellitus. Genetic studies of type 2 diabetes mellitus become increasingly important. The knowledge of the molecular background of type 2 diabetes has been growing rapidly over recent years. One of the forms of the disease defined on the molecular level is maternally inherited type 2 diabetes mellitus. This diabetes, which is frequently accompanied by hearing impairment of deafness (maternally inherited diabetes with deafness-MIDD), was linked with sequence differences in mitochondrial DNA. The most frequent cause of MIDD is A3243G substitution in a mitochondrial tRNA(Leu) gene. While this mutation was identified in different races in several populations, it is still important and valuable to evaluate its prevalence in various ethnic groups. The aim of the project was to determine the prevalence of A3243G substitution in a mitochondrial tRNA(Leu) gene among Polish diabetic subjects. MATERIAL AND METHODS:In total 129 individuals, with type 2 diabetes and 12 with gestational diabetes were selected for this study. Two techniques based on restriction fragment length polymorphism (RFLP) method were used to screen for A3243G mutation. In the first approach, non-radioactive PCR reactions of mitochondrial DNA region of interest were performed using DNA of the study participants. This was followed by Apa I restriction enzyme digestion of the PCR product. Subsequently an electrophoretic separation was done on 2% agarose gel with ethidium bromide staining. In the second, more sensitive, modification of RFLP, [alpha 32P]dCTP was used for internal primer labeling and the electrophoresis was done on acrylamide gel. A positive sample was used to control the quality of the genotyping. RESULTS:In both approaches none of the samples, except for the positive control, showed the evidence of the G variant. CONCLUSIONS:In summary, the A3243G mutation in mitochondrial tRNA(Leu) gene is not a frequent cause of diabetes in the Polish population. Further screening of enlarging study group is necessary to fully determine the prevalence of this mutation in our population. This, together with the search for other mitochondrial mutations, should allow to fully determine the prevalence of MIDD and its specific molecular background in the Polish population.
journal_name
Med Sci Monitjournal_title
Medical science monitor : international medical journal of experimental and clinical researchauthors
Małecki M,Klupa T,Wanic K,Frey J,Cyganek K,Sieradzki Jsubject
Has Abstractpub_date
2001-03-01 00:00:00pages
246-50issue
2eissn
1234-1010issn
1643-3750pii
1669journal_volume
7pub_type
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