Spermatozoa are produced in the testis and undergo post-gonadal modifications in the epididymis to acquire fertilizing ability. In epididymal plasma, high-molecular-weight proteins and such small molecules as free-L carnitine convert the gametes into "competent' and functional cells. This review summarizes the knowledge pertaining to L-carnitine and the significance of free L-carnitine uptake into the mature spermatozoa of mammals. We provide an overview of the function of free L-carnitine and carnitine esters in the metabolism of eukaryotic cells and review the role of the specific carnitine acyltransferases in mitochondrial transport of fatty acids and in modulating acyl-coenzyme A (CoA) pools in cellular organelles. In mammals, including man, free L-carnitine is taken from blood plasma and concentrated in the epididymal lumen. This epididymal secretion is beneficial for spermatozoa and is not merely an excretory waste. The uptake of free L-carnitine into the spermatozoa and its metabolic outcome are discussed first in in-vivo and then in in-vitro situations. Free L-carnitine goes through the sperm plasma membrane by passive diffusion. Free L-carnitine is acetylated in mature spermatozoa only. The excess acetyl-CoA from the mitochondria is probably stored as acetyl-L-carnitine and modulates the reserves of free CoA essential to the function of the tricarboxylic acid cycle. These properties of L-carnitine of buffering CoA in the mitochondrial matrix are known in somatic cells but are accentuated in this study of the male germinal cells. In the future, a precise measurement of the in-vivo and in-vitro concentrations of free CoA and acetyl-CoA in the cellular compartments of immature and mature spermatozoa might complete these data. The relationship between the endogenous pools of free and acetylated L-carnitine and the percentage of progressive sperm motility indicates a more important metabolic function related to flagellar movement. In conclusion, the potential to initiate sperm motility, which takes place in the epididymis, is probably independent of the carnitine system, while the energy properties of acetyl-L-carnitine can only be relevant in situations of "energy crisis'. The uptake of "cytoplasmic' free L-carnitine in mature spermatozoa must be a protective form of mitochondrial metabolism, useful to the survival of this isolated cell.

Catalase activity was determined in human semen by measuring the oxygen burst with a Clark electrode, after H2O2 addition. Significant catalase activities (mean +/- SD) were found in migrated, motile spermatozoa (44 +/- 17 nmoles O2/min/10(8) cells) and in seminal plasma of normozoospermic men (129 +/- 59 nmoles O2/min/ml). It has been demonstrated that seminal catalase originated from prostate; however, its activity was not correlated with the usual prostatic markers (such as citric acid and zinc). Our data suggest a multiglandular function secreted by this organ. The catalase activities measured in seminal samples from asthenozoospermic, infertile men were found lower than those from normozoospermic subjects. The understanding of the relative contribution of the different enzyme systems against O2 toxicity (superoxide dismutase, catalase, glutathione peroxidase) seem to be a priority area of research to understand disturbances of sperm function.

Two groups of men were retrospectively selected according to their observed success in in-vitro fertilization. Seminal and post-migration sperm samples from a low fertilization rate group (less than or equal to 33% cleaved embryos) have been compared to results obtained from a high fertilization rate group (greater than or equal to 66%). It was found that a low mean value of the amplitude of lateral sperm head displacement and an increased percentage of abnormal acrosomes were related to in-vitro fertilization failure. None of the individual sperm factors studied was found to determine in-vitro fertilization success with certainty; only when they were considered in combination was it possible to predict the likelihood of successful in-vitro fertilization of human oocytes.

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Ten framesisec microcinematography ("film"), 1 second timed-exposure photomicrography ("photo"), and laser Doppler velocimetry (LDV) were used to analyse the swimming patterns of human spermatozoa after migration (1 h at 37°C) into an overlying layer of either BWW or Menezo's B2 media. The upper layer of medium was carefully removed and further incubated at 37°C for either 4 h (B2) or 5 h (BWW) and the spcrm motility analysed again. Five experiments were performed using semen from different donors. Film and photo analyses gave the relative incidence of nonprogressive and progressively motile spermatozoa plus, for the progressive spermatozoa, the velocities of progression (Vp) and amplitudes of lateral head displacement (Ah). LDV gave the percentage of motile spermatozoa and the modal instantaneous velocity (Vm). All postmigration sperm populations showed large significant increases in the percentage of motile spermatozoa, with good survival during incubation. The progressive postmigration spermatozoa generally moved with greater Vp and Ah than in the initial seminal plasma-diluted material; Vm was also increased. There were further increases in both Vp and Ah during incubation, but no change in Vm was detected. While the majority of spermatozoa were progressive, some showed a highly active pattern of movement which resulted in no net forward progression. The possible homology between these spermatozoa and the "hyperactivated" motility of capacitated spermatozoa in other mammalian species is discussed. Apparent discrepancies between the three methods used for motility analysis were seen, the possible causes and significances of which are also discussed.

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The initiation of motility and modification of energy metabolism of rat caudal epididymal spermatozoa can be induced by dilution in a saline medium. We have investigated in these cells the relationships between the energy reserve (sperm ATP content measured by bioluminescence) and flagellar movement (high speed videomicrography, 200 frames/sec). A steady state was observed in sperm ATP content, progressive velocity (Vp) and flagellar beat frequency (F) with sperm dilution in a medium with glucose, lactate, pyruvate and acetate substrates after 30 minutes of incubation. Without these substrates, changes in metabolic pathways occurred immediately and initially disturbed the relationship between ATP levels and F, suggesting differences in motility initiation when energy is from an endogenous origin via mitochondrial oxidative phosphorylation. This "energy crisis" was reversed by the addition of substrates to the medium. The three-dimensional flagellar movement observed in the presence of substrates quickly became two-dimensional in their absence. The flagellar beat envelope became more splayed, the mean amplitude of lateral head displacement increased and F decreased. The resulting high flagellar beat efficiency can be compared to that observed during hyperactivation which is a physiological event related to a fall in intracellular ATP level. In both media, the displacement of the flagellum in relation to the wave axis varied sinusoidally. The sine period increased with time when the spermatozoa were incubated in the medium without substrates. These results suggest a gradual slowing-down of the velocity of wave formation in the proximal part of the flagellum.

In the male reproductive tract, very high concentrations (mmol l-1) of free L-carnitine and acetyl-L-carnitine are found in the epididymides, seminal plasma and spermatozoa. It has been reported that the uptake of free L-carnitine by spermatozoa might be related to the epididymal maturation of the sperm membrane, since a greater uptake was found by caput than by cauda spermatozoa in vitro. However, the free L-carnitine concentrations estimated inside the gametes were never greater than those of the surrounding medium. In this study, we investigated the mechanism of transport of free L-carnitine and its ester acetyl-L-carnitine, through the plasma membrane of mature and immature epididymal boar spermatozoa. In vitro, we found a passive diffusion of both compounds to the spermatozoa, whatever the maturation stage. The spermatozoa might progress in the epididymal lumen and accumulate high amounts of free L-carnitine. The active uptake of free L-carnitine occurs only across epididymal mucosa. These results are in agreement with those reported on cells of other organs that exchange pharmacological free L-carnitine concentrations (mmol l-1) by a passive mechanism through the plasma membrane. The acetylation of high amounts of free L-carnitine inside the spermatozoa was found only in caudal spermatozoa. This result suggests that oxidative metabolism (producing acetyl CoA) might be more active in mature cells. The acetyl-L-carnitine added to the incubation medium of boar spermatozoa was hydrolysed. Enzymatic activity of the sperm membrane is low and this may partially explain the low concentrations of acetyl-L-carnitine found in the caudal epididymal plasma.