Oxygen uptake by mitochondria in demembranated human spermatozoa: a reliable tool for the evaluation of sperm respiratory efficiency.
In this work we report a relatively simple and fast method for analysing oxygen consumption and therefore mitochondrial functionality, in individual human ejaculates. This oxygraphic method requires a low number of cells, is highly reproducible and linearly correlates with sperm concentration. Our results have shown that oxygen uptake by mitochondria of demembranated sperm cells from normozoospermic subjects is significantly stimulated by a large set of respiratory substrates and ADP. The respiratory control ratio (RCR) values indicate a good coupling between respiration and phosphorylation by sperm mitochondria and thus a well preserved integrity of the mitochondria themselves. Interestingly, whereas the rates of oxygen uptake, as expected, changed with different sperm concentrations, the RCR values remained constant, thus demonstrating a linear response of the assay. In asthenozoospermic subjects, however, a significant decrease in the sperm respiratory efficiency was found. The results obtained suggest that this method, besides its potential clinical application, could be useful for a deeper understanding of the biochemical properties of sperm mitochondria and their role in ATP production in human spermatozoa.
Ferramosca
A
A
Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, Università del Salento, Lecce, Italy.
Focarelli
R
R
Piomboni
P
P
Coppola
L
L
Zara
V
V
D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D053627:Asthenozoospermia / Q000378:metabolism; D001681:Biological Assay / Q000379:methods*; D019069:Cell Respiration; D004734:Energy Metabolism; D006801:Humans; D007038:Hypotonic Solutions; D008297:Male; D008928:Mitochondria / Q000378:metabolism* / Q000648:ultrastructure; D009997:Osmotic Pressure; D010085:Oxidative Phosphorylation; D010100:Oxygen / Q000378:metabolism*; D010101:Oxygen Consumption; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D013094:Spermatozoa / Q000378:metabolism* / Q000648:ultrastructure; D013997:Time Factors
D007038:Hypotonic Solutions; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D010100:Oxygen