Variation of docosahexaenoic acid content in subsets of human spermatozoa at different stages of maturation: implications for sperm lipoperoxidative damage.
The oxidation of phospholipid-bound docosahexaenoic acid (DHA) has been shown to be one of the major factors that limit the motile life span of sperm in vitro. Sperm samples show high cell-to-cell variability in life span and, consequently, in susceptibility toward lipid peroxidation. Therefore, we postulated that there is also cell-to-cell variability in DHA concentration in human spermatozoa. In this study, the concentration of DHA in subsets of human spermatozoa isolated by a discontinuous Percoll density gradient was determined by gas chromatography. Four subsets of human spermatozoa were isolated using a discontinuous Percoll gradient: fraction 1 was enriched in immature germ cells and immature sperm, fractions 2 and 3 contained, mostly, immature sperm with cytoplasmic droplets, and fraction 4 contained, for the most part, morphologically normal sperm, as determined by histochemical analysis. The results indicated that there were significant differences in DHA content in sperm from all 4 fractions. DHA content in sperm from fraction 1 was 2.5-fold higher than that found in fraction 4. DHA content in mouse sperm obtained from the seminiferous tubules was 3-fold higher than that found in mouse sperm obtained from the epididymis, consistent with the findings observed in ejaculated human sperm. The results of this study indicate (i) there is cell-to-cell variability in the concentration of DHA in human sperm and (ii) that there is a net decrease in DHA content in sperm during the process of sperm maturation.
Ollero
M
M
Department of Obstetrics and Gynecology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA.
Powers
R D
RD
Alvarez
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JG
D000818:Animals; D002499:Centrifugation, Density Gradient; D004281:Docosahexaenoic Acids / Q000378:metabolism*; D006801:Humans; D015227:Lipid Peroxidation; D008055:Lipids / Q000302:isolation & purification; D008297:Male; D051379:Mice; D011205:Povidone / Q000494:pharmacology; D012822:Silicon Dioxide / Q000494:pharmacology; D013094:Spermatozoa / Q000737:chemistry / Q000378:metabolism*; D013997:Time Factors
D008055:Lipids; D004281:Docosahexaenoic Acids; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone