Differentiating male germ cells express a testis-specific form of cytochrome c (Cyt c(T)) that is distinct from the cytochrome c expressed in somatic cells (Cyt c(S)). To examine the role of Cyt c(T) in germ cells, we generated mice null for Cyt c(T). Homozygous Cyt c(T)(-/-) pups were statistically underrepresented (21%) but developed normally and were fertile. However, spermatozoa isolated from the cauda epididymis of Cyt c(T)-null animals were less effective in fertilizing oocytes in vitro and contain reduced levels of ATP compared to wild-type sperm. Sperm from Cyt c(T)-null mice contained a greater number of immotile spermatozoa than did samples from control mice, i.e., 53.1% +/- 13.7% versus 33.2% +/- 10.3% (P < 0.0001) for vas deferens sperm and 40.1% +/- 9.6% versus 33.2% +/- 7.5% (P = 0.0104) for epididymal sperm. Cyt c(T)-null mice often exhibit early atrophy of the testes after 4 months of age, losing germ cells as a result of increased apoptosis. However, no difference in the activation of caspase-3, -8, or -9 was detected between the Cyt c(T)(-/-) testes and controls. Our data indicate that the Cyt c(T)-null testes undergo early atrophy equivalent to that which occurs during aging as a consequence of a reduction in oxidative phosphorylation.

Testis-specific cytochrome c-null mice produce functional sperm but undergo early testicular atrophy

Sodium arsenite induces ATP depletion and mitochondrial damage in HeLa cells

In 146 males aged between 20 years and 40 years attending an infertility service, the secretory activity of the seminal vesicles was assessed by measurement of corrected seminal fructose concentration. This value was related to the presence of a positive semen culture, other evidence of inflammatory processes in the reproductive tract and sperm motility. Only 48% of subjects with a positive semen culture showed evidence of inflammation in the reproductive tract, as assessed by the presence of more than 20 white blood cells per high power field, and greater than 10% spermagglutination in the ejaculate. There was a relationship between the inflammatory process, hypofunction of the seminal vesicles and poor sperm motility. When the semen culture was positive but there was no evidence of inflammation neither seminal vesicle function nor sperm motility was affected. When the semen culture was negative, i.e. no evidence of inflammation and the subjects were asthenozoospermic, the corrected fructose levels were normal. It is proposed that in these conditions the cause of asthenozoospermia may be factors other than accessory sex organ dysfunction. In conclusion, there was no close relationship between the bacteriological results and evidence of inflammation of the accessory glands. A positive semen culture was related to lower levels of corrected fructose (hypofunction of the seminal vesicles) when the positive sperm culture was associated with inflammation of the reproductive tract and asthenozoospermia.

The secretory activity of the seminal vesicles and its relationship to sperm motility: effects of infection in the male reproductive tract

The seminal adenosine triphosphate (ATP) content was determined by bioluminescence after treatment with trichloroacetic acid (TCA) in 81 semen samples 1.5 h after ejaculation obtained from men attending our fertility clinic, and selected to contain either 20% or less spermatozoa with good progressive motility (n = 22), or 60% or more spermatozoa with good progressive motility (n = 59) (Study I), and in 18 semen samples from fertile men 30 min and 3.5 h after ejaculation (Study II). The latter samples were divided into 2 equally large groups according to sperm motility. In Study I the mean sperm ATP concentration was significantly higher in the semen samples with bad motility (0.63 nmol per living spermatozoa x 10(-6)) than in semen samples with good motility (0.39 nmol per living spermatozoa x 10(-6); P less than 0.01). In Study II the ATP concentration per living spermatozoa was also lower in the group with the best motility in comparison with the spermatozoa with lower motility (P less than 0.01), both 30 min and 3.5 h after ejaculation. During the 3-5 h incubation the sperm ATP concentration decreased by 21% (P less than 0.01) in the former group of samples but remained unchanged in the latter group. The results indicate that, in semen samples with highly motile spermatozoa, the consumption of ATP is higher than in semen samples with impaired sperm motility. It is therefore essential that the time between ejaculation and ATP measurement is as short as possible to obtain comparable results. Repeated ATP measurements in combination with an analysis of the number of living spermatozoa, may provide further information on the fertilizing capacity of spermatozoa.

Adenosine triphosphate (ATP) in human spermatozoa

To explore the new mechanisms of tamoxifen (TAM) in the treatment for patients with idiopathic oligoasthenospermia—antioxidation. In a prospective, randomized, controlled clinical trial, 120 cases of idiopathic oligoasthenospermia were enrolled and randomly assigned to the indomethacin group (n = 60) treated with indomethacin (25 mg, bid) and TAM group (n = 60) treated with TAM (10 mg, bid) for 3 months. Before and after treatment, we evaluated semen parameters, serum malondialdehyde (MDA) and total antioxidant capacity (TAC), seminal plasma MDA and TAC, spermatozoa intracellular reactive oxygen species (ROS), sperm succinate dehydrogenase (SDH) activity, sperm mitochondrial membrane potential (MMP), and sperm adenosine triphosphate (ATP) content. The independent t test and one-way repeated measures analysis of variance were used to compare the variables between and within two groups. In the indomethacin group, the percentage of progressive motile sperms, total motility, sperm MMP, and ATP content were increased significantly after 3-month treatment (P < 0.05). In the TAM group, total sperm count, sperm concentration, the percentage of progressive motile sperms, total motility, serum and seminal plasma TAC, sperm MMP, and ATP content were significantly improved or increased (P < 0.05), while spermatozoa intracellular ROS was significantly decreased (P < 0.05). Compared to the indomethacin group, TAM treatment showed better improvement in total sperm count, sperm concentration, serum TAC, seminal plasma TAC, spermatozoa intracellular ROS, and sperm SDH activity. TAM treatment can significantly improve sperm quality, which is achieved through alleviating oxidative stress, improving sperm mitochondrial functionality, and subsequently increasing sperm motility.

Tamoxifen is a potent antioxidant modulator for sperm quality in patients with idiopathic oligoasthenospermia

ATP levels in human spermatozoa have been positively correlated with good motility. This has given rise to the impression that good ATP levels per se are related to good motility, i.e., the higher the ATP level, the better the motility and fertilizing potential. There was no direct correlation between motility percentage, forward progression, viability percentage, and ATP levels (when expressed per 1 million spermatozoa) in the general population. This finding was not unexpected since other factors, such as defects of the microtubules and viscosity of the semen, could affect the motility in some patients. However, when semen from individual patients was assessed, the motility percent, viability, and ATP concentration decreased by comparable levels over a 4-h period postejaculation. Semen samples with normal counts of spermatozoa (<20 × 106/ml) had higher levels of ATP than samples from patients with oligozoospermia. Spermatozoa from patients whose semen contained <0.60 × 10−2 nmol ATP/1 million spermatozo...

Adenosine triphosphate levels in human spermatozoa.

Summary: This study attempts to provide guidelines for the extraction and estimation of ATP from spermatozoa. The conditions of extraction of ATP from spermatozoa using trichloroacetic acid were optimized. The efficiency of this extraction procedure was compared to that using boiling Tris buffer. Three commercially available kits which use the firefly luciferase assay for ATP and two luminometers were evaluated for the measurement of the spermatozoa extracts. Elevated levels of Mg2+ and/or Ca2+ did not interfere with the extration of ATP by trichloroacetic acid or with the subsequent estimation of the nu-cleotide.



Extraktion und Bestimmung von ATP in menschlichen Spermatozoen



Zusammenfassung:  Die vorliegende Untersuchung versucht Richtlinien aufzustellen fur die Extraktion und Bestimmung von ATP in menschlichen Spermatozoen. Die Extrak-tionsbedingungen fur ATP wurden unter Verwendung von Trichloressigsaure optimiert. Die Effizienz des Extraktionsverfahrens wurde mit einem Verfahren verglichen, bei wel-chem siedender Tris-Puffer Verwendung fand. Es wurden drei handelsubliche Kits, welche den Leuchtkafer-Luciferase-Assay fur ATP und zwei Luminometer benutzen, fur die Mes-sung der Spermatozoenextrakte bewertet. Die erhohten Werte von Mg2+ und/oder Ca2+ interferiersen nicht mit der Extraktion von ATP mittels Trichloressigsaure oder mit der nachtraglichen Bestimmung des Nukleotids.

G. Lyons

Papers

Adenosine triphosphate levels in human spermatozoa.

Extraction and Estimation of ATP from Human Spermatozoa