Human Reproduction, Vol. 18, No. 3, 550-556,
March 2003
© 2003 European Society of Human Reproduction and Embryology
Increased sperm mitochondrial DNA content in male infertility
1 INSERM EMI-U 00-18, Laboratoire de Biochimie et Biologie Moléculaire, 2 Laboratoire d’Histologie–Embryologie–Cytologie, UF de Biologie de la Reproduction and 3 Service de Gynécologie, UF d’Assistance Médicale à la Procréation, CHU d’Angers, F-49033 Angers and 4 Biologie de la Reproduction, Pavillon de la Mère et de l’Enfant, CHU de Nantes, BP 1005, F-44093 Nantes cedex 1, France
5 To whom correspondence should be addressed. e-mail: pareynier{at}chu-angers.fr
BACKGROUND: There is increasing evidence that mitochondrial DNA (mtDNA) anomalies in sperm may lead to infertility. Point mutations, deletions and the presence of a specific mtDNA haplogroup have been associated with poor sperm quality, but little attention has been paid to the role of mtDNA content. METHODS: Using density gradient separation and swim-up methods, we selected motile sperm from 32 normal and 35 abnormal sperm samples. The mtDNA/
-globin gene ratio was determined by real-time quantitative PCR. RESULTS: The average mtDNA/-globin ratio of sperm collected from 100% density layers was 1.4 for normal sperm, 6.1 for sperm samples presenting at least one abnormal criterion [among the three criteria established by World Health Organization (1999), i.e. sperm count, motility and morphology], and 9.1 for sperm samples presenting two or more of these abnormal criteria. These differences are very highly significant (P < 0.0001). The mtDNA numbers were also much greater in sperm collected from the 40% density gradient layers (mean: 17.1, P < 0.001), known to contain the most abnormal sperm of the sperm samples, than in those collected from the 100% layers known to contain sperm with the best fertilizing ability. CONCLUSION: Our results showed significant mtDNA amplification in sperm collected from abnormal sperm samples.
Key words: male infertility/mitochondrial DNA/real-time PCR/sperm
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