The spinal motonucleus of the genitofemoral nerve regulating scrotal temperature can also be related to prenatal and neonatal testicular descent by gubernacular change in rats, and a sexually dimorphic-like bulbocavernosus/dorsolateral motonucleus. There is a hypothesis that neonatal androgen affects these motonuclei, and induces development of sexual organs through neural stimulation. Until now, the accumulation of isotope-labelled androgen and the immuno-reactivity of androgen receptor protein in each sexually-dimorphic spinal motonucleus have been revealed in adult rats but they have not been established in rats during neonatal periods. To investigate the presence of the androgen receptor in spinal sexually-dimorphic motonuclei in the neonatal period, we evaluated the androgen receptor immunoreactivity of these motonuclei. In Sprague-Dawley male rats, the lumbar spinal cords were resected at postnatal days 3, 10 and 30, and stained immunohistochemically using polyclonal antibody of androgen receptor protein. The immunoreactivity of androgen receptor protein was observed in the cells of the genitofemoral motonucleus from the 13th thoracic to the 2nd lumbar spinal cord and the bulbocavernosus/dorsolateral motonucleus was observed from the 4th to 5th lumbar spinal cord in all age groups. The proportional areas of both motonuclei at days 3 and 10 on cross-section were larger than at day 30. The motonuclei at days 3 and 10 were similar in all age groups. With the above results, the presence of androgen receptor protein was confirmed in the genitofemoral and bulbocavernosus/dorsolateral motonucleus from neonate to day 30. The larger proportional area of these motonuclei in neonates may indicate an active role for these motonuclei during the neonatal period. Although the immunoreactivity does not directly imply the presence of a functional receptor, neonatal androgen could be responsible for the development of sexual organs through the spinal motonucleus.
Animal ; Animals, Newborn ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, Androgen/immunology ; Receptors, Androgen/analysis* ; Sex Characteristics* ; Spinal Cord/chemistry*

Adolescent ; Androgen-Insensitivity Syndrome ; genetics ; Female ; Humans ; Male ; Mutation ; Polymerase Chain Reaction ; Receptors, Androgen ; genetics ; Sequence Analysis, DNA

To study the X chromesome masaicism in the cytogenetically pure 45,X Turner syndrome patients, we applied PCR technique using DNAs extracted from archived cytogenetic slides. We amplified the DNAs using nested primers targeted to a highly polymorphic short tandem repeat(STR) of the human androgen receptor gene(HUMARA) for the detection of X chromosome mosaicism. This assay is a very sensitive and useful method which can be applied to the DNAs extracted from archived cytohenetic slides to detect X mosaicism. We have tested 50 normal Korean females to determine whether the HUMARA locus is highly polumorphic among Koreans. 85% of Korean population showed heterozygosity in the HUMARA locus. We analysed the 24 DNAs extracted from archived slides of patients and abortuses with Turner syndrome in cytogenetic analysis. We observed the heterozygosities of 50% from pure 45,X patients, 83% from the patients with mosaic Turner syndrome and 8.3% from the abortuses of pure 45,X. Using the PCR tecjhnique of the HUMARA locus in the archived cytogenetic slides, we detected X chtomosome mosaicsm which could not be detected in cytogenetic analysis.
Cytogenetic Analysis ; Cytogenetics* ; DNA* ; Female ; Humans ; Mosaicism* ; Polymerase Chain Reaction ; Receptors, Androgen ; Turner Syndrome* ; X Chromosome

OBJECTIVETo explore the pathogenetic mechanism for a female patient affected with hemophilia A (HA).

METHODSPotential genetic defect was detected with inverse shifting-polymerase chain reaction (IS-PCR). The pattern of X chromosome inactivation was determined with a human androgen receptor assay (HUMARA assay). G-banded karyotyping was carried out to exclude potential chromosome aberrations.

RESULTSIS-PCR showed that the defect of FVIII gene was the distal type of intron 22 inversion. The HUMARA assay showed that the X chromosome inactivation was non-random, and that the mother's X chromosome activity was lower than that of the father's X chromosome which has carried the inverted FVIII gene. No abnormalities were found with G-banded chromosomes.

CONCLUSIONThe prevalence of female HA patient may be caused by non-random inactivation of X chromosomes.

Adolescent ; Female ; Hemophilia A ; etiology ; genetics ; Humans ; Karyotyping ; Polymerase Chain Reaction ; Receptors, Androgen ; analysis ; X Chromosome Inactivation

OBJECTIVETo identify the mutation of human androgen receptor gene (AR) in a patient with complete androgen insensitivity syndrome (CAIS).

METHODSDNA sequences of 8 exons and their exon/intron boundaries of the AR gene in the patient were amplified by PCR and directly sequenced.

RESULTSDNA sequencing revealed a nonsense mutation in exon 1, resulting in a change of codon 441 GAA (glutamic acid) to a stop codon (TAA).

CONCLUSIONA novel mutation Glu441stop (GAA to TAA) of the androgen receptor gene leading to complete androgen insensitivity syndrome was identified in this study in a Chinese patient. It may help us further understanding the pathogenesis of CAIS.

Adult ; Androgen-Insensitivity Syndrome ; genetics ; Base Sequence ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Polymerase Chain Reaction ; methods ; Receptors, Androgen ; genetics ; Sequence Analysis, DNA ; methods

OBJECTIVETo identify potential mutation of androgen receptor (AR) gene in a patient with complete androgen insensitivity syndrome (CAIS) and his family members.

METHODSTotal RNA and genomic DNA were extracted from the peripheral blood samples derived from the proband and her family members. Sequences of 7 exons of the AR gene were amplified with reverse transcriptase PCR(RT-PCR) and subjected to direct sequencing. Suspected mutation was also analyzed with PCR-restriction fragment length polymorphism (PCR-RFLP) and direct sequencing.

RESULTSDNA sequencing has revealed a nucleotide change (2880A>G) in the pedigree, which resulted in a missense mutation (R840H).

CONCLUSIONA prenatal diagnostic method was established for detecting mutation of the AR gene in the pedigree. Long chain RT-PCR was first used for the detection of AR gene mutations.

Androgen-Insensitivity Syndrome ; genetics ; Base Sequence ; Child ; DNA Mutational Analysis ; methods ; Family Health ; Female ; Humans ; Male ; Mutation, Missense ; Pedigree ; Receptors, Androgen ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods

Androgen insensitivity syndrome (AIS), an X-linked recessive genetic disorder of sex development, is caused by mutations in the androgen receptor (AR) gene, and is characterized by partial or complete inability of specific tissues to respond to androgens in individuals with the 46,XY karyotype. This study aimed to investigate AR gene mutations and to characterize genotype-phenotype correlations. Ten patients from unrelated families, aged 2-31 years, were recruited in the study. Based on karyotype, altered hormone profile, and clinical manifestations, nine patients were preliminarily diagnosed with complete AIS and one with partial AIS. Genetic analysis of AR gene revealed the existence of 10 different mutations, of which five were novel (c.2112 C>G[p.S704R], c.2290T>A[p.Y764N], c.2626C>T[p.Q876X], c.933dupC[p.K313Qfs*28], and c.1067delC[p.A356Efs*123]); the other five were previously reported (c.1789G>A[p.A597T], c.2566C>T[p.R856C], c.2668G>A[p.V890M], c.2679C>T[p.P893L], and c.1605C>G[p.Y535X]). Regarding the distribution of these mutations, 60.0% were clustered in the ligand-binding domain of AR gene. Exons 1 and 8 of AR gene each accounted for 30.0% (3/10) of all mutations. Most of the truncation mutations were in exon 1 and missense mutations were mainly located in exons 4-8. Our study expands the spectrum of AR gene mutations and confirms the usefulness of AR gene sequencing to support a diagnosis of AIS and to enable prenatal or antenatal screening.
Adolescent ; Adult ; Androgen-Insensitivity Syndrome/genetics* ; Child ; Child, Preschool ; DNA Mutational Analysis ; Genetic Association Studies ; Humans ; Male ; Mutation, Missense ; Phenotype ; Receptors, Androgen/genetics* ; Symptom Assessment ; Young Adult

OBJECTIVESTo study the distribution and potential function of androgen receptor (AR) mRNA and AR in rat submaxillary.

METHODSIn situ hybridization using digoxigenin-labled oligonucleotide probes, cell culture and radio-immunoassay were performed to localize the AR and detect the concentration of epidermal growth factor (EGF) in culturing supernant.

RESULTSAR mRNA hybridization signals were detected in glandular epithelial cells of serous acinus and epithelial cells in all gland ducts. The signals distributed in cytoplasma of all positive cells with negative nuclei; Administration of testosterone can significantly increase the level of EGF (P < 0.05).

CONCLUSIONSThe rat submaxillary not only is a target organ of androgen but also can product AR by itself. When androgen combined with androgen receptor and can submaxillary function can be infected and can result in the elevation of the level of EGF secreted.

Animals ; Epidermal Growth Factor ; biosynthesis ; Male ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Receptors, Androgen ; genetics ; physiology ; Submandibular Gland ; metabolism

OBJECTIVETo investigate the expression of cyclin D1, p16, AR and ER in fibroblasts of scars for further understanding the interaction of these factors and the roles that they play in scar development.

METHODSThirty samples of mature scar, hypertrophic scar and keloid were detected with immunohistochemical method (SP technique) and compared with normal skin.

RESULTSThere were on positive results in normal skin and mature scars. The expression of cyclin D1, p16 and AR was higher in hypertrophic scars and keloids than in normal skin with significant difference (P < 0.05). The expression of cyclin D1 in keloids was higher than in hypertrophic scars (P < 0.05). Though the expression of p16 was higher in keloids than in hypertrophic scars, the difference was not significant. There was significant correlation between the expression of cyclin D1 and AR in the pathologic scar.

CONCLUSIONThe AR played an important role in scar formation and displayed its function through cyclin D1. The expression of p16 could suppress the excessive proliferation of cells to some extent. If the effect was not enough to resist the function of cyclin D1, long-term proliferation of cells would occur and lead to keloid formation. As the expression of cyclin D1 and p16 in hypertrophic scars was in a state of relative equilibrium, the cell proliferation showed a tendency of self-restriction.

Cicatrix ; metabolism ; pathology ; Cyclin D1 ; analysis ; metabolism ; Cyclin-Dependent Kinase Inhibitor p16 ; analysis ; metabolism ; Fibroblasts ; chemistry ; metabolism ; Humans ; Immunohistochemistry ; Receptors, Androgen ; analysis ; metabolism ; Receptors, Estrogen ; analysis ; metabolism ; Skin ; chemistry ; metabolism ; pathology

Androgen insensitivity syndrome (AIS) is a X-linked disorder of sexual differentiation resulting from defective androgen receptor (AR) function. Androgens are secreted by the testes of 46,XY individuals, but there is loss of target organ response to the hormone. The abnormalities of AR are due to defects in the AR gene, and many mutations causing AIS have been reported since the cloning of AR gene. In this study, we analyzed the AR genes in twelve Korean patients with androgen insensitivity syndrome: 9 patients with complete AIS and 3 patients with partial AIS DNAs were isolated from patients with AIS, and the coding region of AR gene was amplified by a polymerase chain reaction using 7 pairs of primers (exon B-H). Sequence analysis of the AR gene was performed using direct sequencing and single strand conformational polymorphism (SSCP). The AR gene mutations were identified in 7 out of 12 patients: 6 of 9 patients with complete AIS, and one of 3 patients with partial AIS. Mutations found were as follows: the point mutation (ATT->ACT) at position 680 of exon D, point mutation (TGG->TGC) at position 751 of exon E, point mutation (CAA->TAA) at position 792 of exon F, point mutations (CGC->TGC, GTG->ATG) at position 855 and 866 of exon G, and the deletion of 13 nucleotides (CGTATCATTGCAT) at position 840 of exon G, respectively. To the best of our knowledge, the point mutations found in exon D, exon E, and exon F, and the deletion in exon G have not been observed before. SSCP revealed bands with abnormal mobility in 10 out of 12 patients tested. Mutations were found 5 out of these 10 patients. The other two patients showed no abnormal band on SSCP, but showed mutations by direct sequencing. In conclusion, we have demonstrated the AR gene mutations, including three novel mutations, in Korean patients with AIS, and these abnormalities might be related to the pathogenesis of androgen insensitivity syndrome.
Androgen-Insensitivity Syndrome* ; Androgens ; Clinical Coding ; Clone Cells ; Cloning, Organism ; DNA ; Exons ; Humans ; Male ; Nucleotides ; Point Mutation ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Receptors, Androgen* ; Sequence Analysis ; Sex Differentiation ; Testis