Human Molecular Genetics Advance Access originally published online on October 17, 2006
Human Molecular Genetics 2006 15(23):3411-3419; doi:10.1093/hmg/ddl417
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Non-invasive genetic diagnosis of male infertility using spermatozoal RNA: KLHL10mutations in oligozoospermic patients impair homodimerization
1 Department of Pathology, 2 Department of Molecular and Human Genetics, 3 Department of Molecular and Cellular Biology, and 4 Scott Department of Urology, Baylor College of Medicine, Houston, Texas 77030, USA and 5 Department of Physiology and Cell Biology, University of Nevada School of Medicine, Reno, Nevada, 89557, USA
* To whom correspondence should be addressed at: Department of Pathology One Baylor Plaza, Baylor College of Medicine, Houston, Texas 77030, USA. Tel: +1 7137986451; Fax: +1 7137985833; Email: mmatzuk{at}bcm.tmc.edu
Received August 28, 2006; Accepted October 13, 2006
Infertility affects an estimated 7% of men worldwide, nearly a quarter of whom are diagnosed as idiopathic. The genetic etiologies of idiopathic male infertility are unknown, partly due to lack of simple diagnostic techniques. Moreover, the transmission risk of such genetic defects to offspring born from assisted reproductive techniques is increasingly becoming a concern for physicians and infertile couples. We explored the feasibility of obtaining full-length mRNAs from transcriptionally inert human spermatozoa in semen as a non-invasive diagnostic tool for identifying germline mutations in candidate infertility-associated genes. The efficacy of reverse-transcription PCR on spermatozoal RNA from infertile patients with wide-ranging sperm concentrations varied between 91 and 99% for multiple haploid germ cell-expressed genes. Using this methodology, we identified seven oligozoospermic patients with missense and splicing mutations in the germ cell-specific gene, KLHL10. Three of 270 (1.1%) severely oligozoospermic patients (<106 sperm/ml) harbor KLHL10 alterations that were absent in 394 controls and exhibited significant association (P=0.02). Two KLHL10 missense mutations (A313T and Q216P) resulted in impaired homodimerization with the wild-type protein in yeast interaction assays, suggesting a functional deficiency. This study demonstrates the utility of this approach for analysis of haploid germ cell-expressed genes regulating post-meiotic events including sperm maturation, motility and fertilization. The development of non-invasive techniques to analyze genetic defects of human spermatogenesis, previously possible only with invasive testis biopsies, provides important diagnostic and therapeutic implications for reproductive medicine.
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors.
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