AIMTo study the occurrence of Y chromosome microdeletions in azoospermic patients with Klinefelter's syndrome (KFS).

METHODSBlood and semen samples were collected from azoospermic patients with KFS (n = 14) and a control group of men of proven fertility (n = 13). Semen analysis was done according to World Health Organization (WHO) guidelines. Blood samples were processed for karyotyping, fluorescent in situ hybridization (FISH) and measurement of plasma follicle stimulating hormone (FSH) by radioimmunoassay. To determine Y chromosome microdeletions, polymerase chain reaction (PCR) of 16 sequence tagged sites (STS) and three genes (DFFRY, XKRY and RBM1Y) was performed on isolated genomic DNA. Testicular fine needle aspiration cytology (FNAC) was done in selected cases.

RESULTSY chromosome microdeletions spanning the azoospermia factor (AZF)a and AZFb loci were found in four of the 14 azoospermic patients with KFS. Karyotype and FISH analysis revealed that, of the four cases showing Y chromosome microdeletion, three cases had a 47,XXY/46,XY chromosomal pattern and one case had a 46,XY/47,XXY/48,XXXY/48,XXYY chromosomal pattern. The testicular FNAC of one sample with Y chromosome microdeletion revealed Sertoli cell-only type of morphology. However, no Y chromosome microdeletions were observed in any of the 13 fertile men. All patients with KFS had elevated plasma FSH levels.

CONCLUSIONPatients with KFS may harbor Y chromosome microdeletions and screening for these should be a part of their diagnostic work-up, particularly in those considering assisted reproductive techniques.

Adolescent ; Adult ; Chromosome Deletion ; Chromosomes, Human, Y ; Electrophoresis, Gel, Two-Dimensional ; Genetic Loci ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Klinefelter Syndrome ; complications ; genetics ; Male ; Mosaicism ; Oligospermia ; etiology ; genetics ; Seminal Plasma Proteins ; genetics ; Sequence Tagged Sites

AIMTo identify the genotype of two Indians with male pseudohermaphroditism.

METHODSStandard radioimmunoassay procedure was used for estimating hormonal levels. Conventional cytogenetic analysis was carried out for diagnosing the genetic sex in these subjects with genital ambiguity. Molecular analysis was carried out by standard polymerase chain reaction procedure using different sets of primers and reaction conditions specific for the 5alpha-reductase type 2 gene (SRD5A2) gene. Direct sequencing was carried out using the ABI Prism dye terminator sequencing kit and the ABI 310 sequencing apparatus.

RESULTSWe found an SRD5A2 gene mutation in exon 5, where arginine is substituted with glutamine (R246Q), in two males with pseudohermaphroditism and ambiguous genitalia from unrelated families. This is the first time this mutation has been reported in individuals from India.

CONCLUSIONIdentification of the R246Q mutation of the SRD5A2 gene from two unrelated Indian families possibly extends the founder gene effect.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase ; genetics ; Child ; Dihydrotestosterone ; blood ; Disorders of Sex Development ; genetics ; pathology ; Family Health ; Follicle Stimulating Hormone ; blood ; Founder Effect ; Genitalia, Male ; abnormalities ; Humans ; Hypospadias ; genetics ; pathology ; India ; Luteinizing Hormone ; blood ; Male ; Mutation, Missense ; Testosterone ; blood

AIMTo determine if Yq microdeletion frequency and loci of deletion are similar in two tissues (blood and sperm) of different embryological origin.

METHODSThe present study included 52 infertile oligozoospermic cases. In each case, DNA was isolated from blood and sperms and polymerase chain reaction (PCR) microdeletion analysis was done from genomic DNA isolated from both the tissues. The PCR products were analyzed on a 1.8% agarose gel. PCR amplifications found to be negative were repeated at least three times to confirm the deletion of a given marker.

RESULTSOnly 1 case harbored microdeletion in blood DNA, whereas 4 cases harbored microdeletion in sperm DNA.

CONCLUSIONThe frequency of Yq microdeletions is higher in germ cells as compared to blood. As the majority of infertile couples opt for assisted reproduction procreation techniques (ART), Yq microdeletion screening from germ cells is important to understand the genetic basis of infertility, to provide comprehensive counseling and most adapted therapeutics to the infertile couple.

Chromosomes, Human, Y ; genetics ; DNA ; blood ; genetics ; isolation & purification ; Humans ; Male ; Repetitive Sequences, Nucleic Acid ; Sequence Deletion ; Spermatozoa ; physiology