Human uniparental gestations such as gynogenetic ovarian teratomas provide a model to evaluate the integrity of parent-specific gene expression - i.e. imprinting - in the absence of a complementary parental genetic contribution. The few imprinted genes characterized so far include the insulin-like growth factor-2 gene (IGF2) coding for a fetal growth factor and H19 gene whose normal function is unknown but it is likely to act as an mRNA. IGF2 is expressed by the paternal allele and H19 by the maternal allele. This reciprocal expression is quite interesting because both H19 and IGF2 genes are located close to each other on chromosome 11p15.5. In situ RNA hybridization analysis has shown variable expression of the H19 and IGF2 alleles according to the tissue origin in 11 teratomas. Especially, Skin, derivative of ectoderm, is expressed conspicuously. We examined imprinting of H19 and IGF2 in teratomas using PCR and RT-PCR of exonic polymorphism. H19 and IGF2 transcript could be expressed either biallelically or monoallelically in the teratomas. Biallelic expression (i.e., loss of imprinting) of IGF2 occured in 5 out of 6 mature teratomas and 1 out of 1 immature teratoma. Biallelic expression of H19 occured in 4 out of 10 mature teratomas and 1 out of 1 immature teratoma. Expression levels of H19 and IGF2 transcript using the semi-quantitative RT-PCR had no relation between monoallelic and biallelic expression. Moreover, IGF2 biallelic expression did not affect allele-specificity or levels of H19 expression. These results demonstrate that both genes, H19 and IGF2, can be imprinted, expressed and regulated independently and individually of each other in ovarian teratoma.
Alleles ; Clinical Coding ; Ectoderm ; Exons ; Fetal Development ; Gene Expression ; Genomic Imprinting ; Humans ; Insulin-Like Growth Factor II* ; Parents ; Polymerase Chain Reaction ; RNA ; RNA, Messenger ; Skin ; Teratoma*

Duchenne and Becker muscular dystrophy are the major neuromuscular disorders with X-linked recessive inheritance. Preimplantation sex determination has been generally used to avoid pregnancies with these diseases. However, in order to determine if the embryo is normal, carrier or affected regardless of the sex, there is a need for a combined analysis of specific exon on dystrophin gene as well as sex determination of embryo using the same biopsied blastomere. If the exon deletion is not determinable, further diagnosis of carrier or patient can be performed by haplotype analysis. In this study, we applied the primer extension preamplification (PEP) method, which amplifies the whole genome, in 40 cases of single amniocyte and 40 cases of chorionic villus cell. We analysed haplotypes using two (CA)n dinucleotide polymorphic markers located at the end of 5' and 3' region of the dystrophin gene. Exon 46 of dystrophin gene and DYZ3 on chromosome Y were chosen as a target sequence for coamplification PCR. Upon optimizing the conditions, the amplification rates were 91.25% (73/80) for haplotypes (92.5% in amniocyte, 90% in chorionic villus cell) and 88.75% (71/80) for coamplification (85% in amniocyte, 92.5% in chorionic villus cell). The result of the study indicates that haplotypes and coamplification using PEP can be applied to prenatal and preimplantation diagnosis in Duchenne muscular dystrophy making it possible to determine if the fetus is a carrier or an affected one.
Blastomeres ; Chorionic Villi ; Diagnosis ; Dystrophin* ; Embryonic Structures ; Exons ; Fetus ; Genome ; Haplotypes* ; Humans ; Muscular Dystrophies ; Muscular Dystrophy, Duchenne ; Polymerase Chain Reaction* ; Pregnancy ; Preimplantation Diagnosis ; Wills

The N-methylpurine-DNA glycosylase (MPG), a ubiquitous DNA repair enzyme, removes N-methylpurine and other damaged purines induced in DNA. Tissue-specific mRNA levels of the N-methylpurine-DNA glycosylase (MPG) were investigated in Balb/c mice of four different growing stages; newborn, 1, 4 and 8-weeks postpartum. MPG expressions in the newborn and the 8-week-old mice were the highest in thymus and testis, respectively. The tested tissues of the newborn mice had consistently higher MPG mRNA level than 8-week-old adults except in testis and thymus. The MPG mRNA level in testis was the lowest in the newborn mice, but it attained the highest in the 8-week-old mice. The levels of MPG mRNA among the different tissues in the newborn and the 8-week-old mice were more than 9.0 and 19.0-fold respectively. These results suggest that the of MPG expression was dependent on the growing stage and had tissue-specificity.
Adult ; Animals ; DNA Repair* ; DNA* ; Gene Expression ; Humans ; Infant, Newborn ; Mice* ; Postpartum Period ; Purines ; RNA, Messenger ; Testis ; Thymus Gland

Previously, we made a study report on the genotype disturbution and the gene frequency of angiotensin I-converting enzyme (ACE) in Korean population, and on the association between hypertension and genetic variance of ACE. This time, we have investigated a rapid mismatch-PCR/RFLP assays for the variant of the angiotensin II type 1 receptor (AT1R) gene (an A C transversion at position 1166 of AT1R gene), a mutation which may interact with the ACE polymorphism in the determining of risk of myocardial infarction. The genotype distributions of Koreans' angiotensin II type I receptor gene were AA (66.3%):AC (28.1%):CC (5.6%), thus the AA genotype was most numerous, and the allele frequency was A:C = 0.803:0.197. Genotype distributions were shown as AA (76.8%):AC (20.9%):CC (2.3%), the allele frequency was A:C = 0.872:0.128 in the male group, and AA (47.4%):AC (41.0%):CC (11.6%), A:C = 0.679:0.321 in the female group. Differences were highly significant between the male and female groups (p<0.0001). Genotype distributions between angiotensin II type I receptor gene and angiotensin converting enzyme gene showed that there is no significance between AT1R genotypes and ACE genotypes in total subjects (p>0.05).
Angiotensin II* ; Angiotensins* ; Female ; Gene Frequency ; Genotype ; Humans ; Hypertension ; Male ; Myocardial Infarction ; Peptidyl-Dipeptidase A* ; Polymorphism, Genetic ; Receptor, Angiotensin, Type 1*

Facioscapulohumeral muscular dystrophy (FSHD) is an autosomal dominant neuromuscular disorder which has been clinically shown to cause progressive weakness and result in atrophy of the facial muscles, shoulder girdle and upper arm muscles. The responsible gene for the FSHD has been located on chromosome 4q35-qter. The probes p13E-11 and pFR-1 detect DNA rearrangements associated with FSHD as under 28 kb DNA fragment in genomic southern analysis digested with EcoR I and the fragment contains 3.3 kb Kpn I tandem repeats. In this study, 4 fetuses with a family history of FSHD were analysed by genomic southern hybridization analysis with probes to determine whether they carried the deleted region. Of the 4 fetuses, three of them had mothers who were FSHD patients and the other one had a father affected with FSHD. After 10-11 weeks of gestation, we performed chorionic villi sampling and extracted DNA from uncultured and cultured tissue cells for the direct DNA analysis. The result of the southern analysis showed two fetuses having received about 15-18 kb of deleted genes from the father and the mother respectively, and found to be FSHD patients. The other two fetuses were shown to have two normal alleles from the parents and found to be normal. Two pregnancies which were determined to be normal were carried to term delivering two healthy babies.
Alleles ; Arm ; Atrophy ; Chorionic Villi Sampling ; DNA* ; Facial Muscles ; Fathers ; Female ; Fetus ; Gene Rearrangement ; Humans ; Mothers ; Muscles ; Muscular Dystrophy, Facioscapulohumeral* ; Parents ; Pregnancy ; Prenatal Diagnosis* ; Shoulder ; Tandem Repeat Sequences

The Lesch-Nyhan syndrome which is caused by the deficiency of hypoxanthine guanine phosphoribosyltransferase is an X-linked recessive disorder characterized by hyperuricemia, choreoathetosis, mental retardation and compulsive self-injurious behavior. Clinical management of the patients with the Lesch-Nyhan syndrome is frustrating and requires burdensome medical treatment since it cripples the patient and shortens the life span by progression of neurological symptoms, but there are no cures or measures for relieving relentless natural course of the disease yet. Therefore, prenatal diagnosis of the affected fetus is important in genetic counselling for the family at high risk. In this study, four different mutations in the HPRT gene of four probands have been identified in four unrelated families; K215X, Q109X, nt.631 A, and nt.289 GT. Two mutations among them altered restriction enzyme sites; SpeI for Q109X and MaeI for nt.289 GT. Based on their molecular defects, prenatal diagnoses of 3 the fetuses were successfully made between ninth and eleventh week of gestation by polymerase chain reaction(PCR), restriction digestion and DNA sequencing using cDNA obtained from chorionic villus samples (CVS). We predicted the outcome of all fetuses prenatally. Among the three fetuses two were male and one was female according to the identification made by PCR amplification of the sex determining region of the Y chromosome(SRY) gene. Each carried a wild type allele for the corresponding mutant allele. They were also tested postnatally for the mutations to be unaffected.
Alleles ; Chorionic Villi ; Digestion ; DNA, Complementary ; Female ; Fetus* ; Humans ; Hyperuricemia ; Hypoxanthine Phosphoribosyltransferase ; Intellectual Disability ; Lesch-Nyhan Syndrome* ; Male ; Polymerase Chain Reaction ; Pregnancy ; Prenatal Diagnosis ; Self-Injurious Behavior ; Sequence Analysis, DNA

This paper reports 3 cases with 46,XX sex reversed male. Three 46,XX hypogonadal subjects showed complete sex reversal and had normal phallus and azoospermia. We studied them under clinical, cytogenetic and molecular aspects to find out the origin of the sex reversal. Patients had markedly elevated serum follicle-stimulating hormone (FSH) and lutenizing hormone (LH) and decreased or normal range of serum testosterone. The testicular volumes were small (3-8ml). Testicular biopsy showed Leydig cell hyperplasia and atrophy of seminiferous tubules. We obtained the results of normal 46,XX and XY dual fluorescent in situ hybridization (FISH) which could rule out the presence of Y chromosome mosaicism. By using polymerase chain reaction (PCR), we amplified short arm (SRY, PABY, ZFY and DYS14), centromere (DYZ3), and heterochromatin (DYZ1) region of the Y chromosome. PCR amplification of DNA from these patients showed the presence of the sex-determining region of the Y chromosome (SRY) but didn't show the centromere and heterochromatin region sequence. The SRY gene was detected in all the three patients. Amplification patterns of the other regions were different in these patients; one had four amplified loci (PABY+, SRY+, ZFY+, DYS14+), another had two loci (SRY+, ZFY+) and the other had two loci (PABY+, SRY+). We have found that each patient's translocation elements had different breakpoints at upstream and downstream of the SRY gene region. We conclude that the testicular development in 46,XX male patients were due to insertion or translocation of SRY gene into X chromosome or autosomes.
Arm ; Atrophy ; Azoospermia ; Biopsy ; Centromere ; Cytogenetics* ; DNA ; Follicle Stimulating Hormone ; Genes, sry ; Heterochromatin ; Humans ; Hyperplasia ; In Situ Hybridization, Fluorescence ; Male* ; Mosaicism ; Polymerase Chain Reaction ; Reference Values ; Seminiferous Tubules ; Testosterone ; X Chromosome ; Y Chromosome

The present report describes a case that showed a normal fetal karyotype in an antenatal genetic study but an abnormal placental karyotype of 46,XX,r(15) on postnatal examination. The pregnancy was complicated by fetal nuchal translucency in the first trimester and intrauterine growth restriction in the second and third trimesters. A 1780 gm female baby was born after 40 weeks of gestation, but died of respiratory distress and sepsis on the 10th day of life. Our case was unique in that the placental chromosomal aberration was a structural abnormality instead of a numerical aberration that is seen in most reported cases of confined placental mosaicism.
Chromosome Aberrations ; Female ; Humans ; Karyotype ; Mosaicism* ; Nuchal Translucency Measurement ; Pregnancy ; Pregnancy Trimester, First ; Pregnancy Trimester, Third ; Sepsis

A female hemihypertrophy patient with hypomelanosis of Ito is presented as a rare case combining classical features of both the syndrome. Chromosomal profile has been based on longitudinal study of repeated lymphocyte cultures during 1984-1992. The propositus has exhibited chromosomal mosaicism both hypoploid (42+/-1) and hyperploid (48+/-2 chromosome) counts, but the major stem line presented 46XX chromosomes. Ring chromosome with simple and complex translocations with marker dots appear to be the major cytogenetic assemblage of this child to posses unequal left and right halves of the body. Each and every organ from toe to the head has grown up unequally and lately the patient had been exhibiting different dark and light shapes of melanin on the skin. We believe that the patient had inherited, through her male parent, "a few" mutated loci on some chromosomes so as to generate different cell lines within the developing child. All sibs and the mother showed normal karyotype with no apparent aberration.
Cell Line ; Child ; Cytogenetics ; Female ; Head ; Humans ; Hypopigmentation* ; Karyotype ; Longitudinal Studies ; Lymphocytes ; Male ; Melanins ; Mosaicism ; Mothers ; Parents ; Ring Chromosomes ; Skin ; Toes

Though Fragile X syndrome is one of the most common inherited causes of mental retardation, it is not much detected yet in Korean population. One of the reason may be that the syndrome is not well known to the special education teachers as well as to the clinicians in this country. Thus, molecular test was undertaken to screen out fragile X syndrome in 122 children of two Korean schools for emotionally severely handicapped children. The subjects were all boys, previously known as having pervasive developmental disorder with or without mental retardation. Southern blot analysis of peripheral blood showed the abnormally enlarged (CGG)n repeat sequence associated with fragile X syndrome in two children. This finding suggests that the DNA testing for fragile X syndrome is warranted for Korean high risk population and that more concern about this syndrome is needed for the professionals who work for mentally handicapped children. The issues involved in genetic counseling for fragile X syndrome are discussed.
Blotting, Southern ; Child ; Disabled Children* ; Disabled Persons* ; DNA* ; Education, Special ; Fragile X Syndrome* ; Genetic Counseling ; Humans ; Intellectual Disability ; Korea* ; Mentally Disabled Persons