Evaluation of mitochondrial respiratory efficiency during in vitro capacitation of human spermatozoa.
The role of mitochondria in sperm motility was the subject of several investigations. However, different views on this topic emerged among scientists. In particular, very little is known on the mechanisms of energy production occurring during human sperm capacitation and related processes. In this study, we have investigated the mitochondrial respiratory efficiency in human sperm samples from normozoospermic subjects before and after swim-up selection and incubation under capacitating condition. Sperm cells, selected by swim-up treatment, were incubated up to 24 h and then demembranated by hypotonic swelling at selected times. The oxygen uptake rate was measured in both basal and swim-up selected samples by a polarographic assay. Mitochondria of swim-up selected cells showed an impressive oxygen consumption rate, which was about 20 times higher than that measured in basal samples. The high mitochondrial respiratory efficiency remained stable up to 24 h after the swim-up treatment. The respiration control ratio, the substrate specificity and the inhibitor sensitivity in the swim-up selected samples were similar to those of basal samples thereby suggesting that the physiology of mitochondria was preserved after the swim-up treatment. Furthermore, the remarkably high mitochondrial respiration in swim-up selected samples allowed the oxygraphic analysis of just 200,000 sperm cells. Sperm selection and incubation under capacitating condition are therefore associated with a high activity of the mitochondrial respiratory chain. The sperm oxygen consumption rate could be useful to exclude mitochondria malfunctioning in male infertility.
Stendardi
A
A
Applied Biology Section, Department of Biomedical Sciences, University of Siena, Siena, Italy.
Focarelli
R
R
Piomboni
P
P
Palumberi
D
D
Serafini
F
F
Ferramosca
A
A
Zara
V
V
D015153:Blotting, Western; D019069:Cell Respiration; D004591:Electrophoresis, Polyacrylamide Gel; D004734:Energy Metabolism; D006801:Humans; D008297:Male; D008928:Mitochondria / Q000378:metabolism*; D010101:Oxygen Consumption; D010766:Phosphorylation; D013075:Sperm Capacitation / Q000502:physiology*; D013081:Sperm Motility / Q000502:physiology; D013094:Spermatozoa / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism
D014443:Tyrosine