Multilocus analyses of estrogen-related genes reveal involvement of the ESR1 gene in male infertility and the polygenic nature of the pathology.

OBJECTIVE: To examine whether polymorphisms within the ESR1, FSHR, ESR2, CYP19A1, and NRIP1 genes are susceptibility factors for human male idiopathic infertility and to test the joint effects of these genes on male reproductive function. DESIGN: Genetic association study of male infertility with polymorphisms, using both single-gene and multilocus approaches. SETTING: Private and public fertility units and a private center for biomedical research. PATIENT(S): One hundred four Spanish men with azoospermia or severe oligozoospermia and 95 unselected race-matched healthy controls from the same geographic region. INTERVENTION(S): Peripheral blood extraction, DNA purification, and ESR1 g.938T>C, FSHR Ser680Asn, ESR2 *39A>G, CYP19A1 *19C>T, and NRIP1 Gly75Gly polymorphism analyses. MAIN OUTCOME MEASURE(S): Single-gene statistical analyses and multilocus statistical analyses with Sumstat, Permutation and Model-free analysis, and Estimating Haplotypes software. RESULT(S): We observed an excess of homozygous infertile men for the ESR1 g.938T>C marker. Multilocus analyses detected genetic interaction between the five candidate gene markers that are influential over male infertility. In addition, we detected a five-loci protector genetic pattern with a frequency of 9.4% in controls but absent in infertile men. CONCLUSION(S): Our results support a relevant role for the estrogenic pathway, notably the ESR1 gene, in human male reproductive function and advocate a complex trait model for male infertility.

Absence of de novo Y-chromosome microdeletions in male children conceived through intracytoplasmic sperm injection.

Molecular screening for Y-chromosome microdeletions in 96 Spanish male children conceived through intracytoplasmic sperm injection (ICSI) was conducted. No microdeletions were detected; these results support the notion that de novo Y-chromosome alterations are rare and unrelated to the ICSI technique itself.

Scanning of Y-chromosome azoospermia factors loci using real-time polymerase chain reaction and melting curve analysis.

OBJECTIVE: To develop a novel method to scan AZF loci looking for microdeletions. DESIGN: Molecular method development. SETTING: Men undergoing reproductive techniques in a private fertility unit. Molecular methods were performed in a private center for biomedical research. PATIENT(S): : Fifty-eight men divided in two groups depending on seminal analyses. A group of 19 women were also included as positive controls (absence of amplification). INTERVENTION(S): Peripheral blood extraction and DNA purification. MAIN OUTCOME MEASURE(S): Our method is based on real-time polymerase chain reaction (PCR) and melting curve analysis. We performed the screening of 16 selected sequence tagged sites (STS) within AZF loci, and we also calculated the mean, range, and standard deviation for melting temperature patterns and the crossing points values for each STS tested. RESULT(S): We detected one azoospermic patient with several STS deleted within the AZFc region. No deletions were detected in a group of 13 healthy men, and no amplification for any of the STS tested were observed in the positive control group (19 healthy women). CONCLUSION(S): We have developed a novel method based on real-time PCR and melting curve analysis to scan AZF loci looking for microdeletions This method is fast and reliable and permits the scanning of DNA from one patient per hour, minimizing the risk of cross contamination, and false-positive and false-negative results.