carbocyanines and Infertility--Male

Sperm motility is the major decisive factor in determining male fertility. The objective of the present study was to analyse the effect of mitochondrial membrane potential (MMP) on the temporal regulation of sperm motility. Observations were recorded in various rodent species and among differentially motile sperm fractions including swim up and leftover layer of human semen sample using JC-1 stain (a marker of the MMP) through FACS. Swim-up sperms having highest motility showed significantly higher MMP as compared to leftover sperms, which had the least motility. Interestingly, infertile patients with compromised motility showed low MMP as compared to the healthy individuals. Further, as per the time lapse, sperm motility goes down, at the same time, it was observed that MMP also decreases in human as well as in rodent sperms. Treatment of known spermicides on human sperms reduced their motility drastically which in turn also reduced its MMP significantly. Treatment of human sperms with oxidative uncoupler also impeded their motility by reducing MMP, indicating a definitive role on MMP on sperm motility and fertility. Based on the results of the study, MMP can be considered as a potential regulator and indicator of sperm motility and hence could be directly related to male fertility.

Topics: Animals; Benzimidazoles; Carbocyanines; Carbonyl Cyanide m-Chlorophenyl Hydrazone; Detergents; Humans; Infertility, Male; Male; Membrane Potential, Mitochondrial; Mice; Oxidation-Reduction; Rats, Sprague-Dawley; Sperm Motility; Spermatocidal Agents; Spermatozoa; Time-Lapse Imaging; Uncoupling Agents

The aim of the study was to examine an in vitro effect of the three bacterial strains (Escherichia coli, Staphylococcus haemolyticus and Bacteroides ureolyticus) on ejaculated spermatozoa with reference to sperm membrane integrity and mitochondrial activity. The study was carried out on swim-up-separated spermatozoa from 12 normozoospermic volunteers. Sperm plasma membrane stability was evaluated by the LIVE/DEAD Sperm Viability Kit and by the merocyanine 540 test. Mitochondrial activity was evaluated using the JC-1 test as well as the NADH-dependent NBT assay. The percentage of dead cells was significantly higher in spermatozoa treated with B. ureolyticus as compared to that of control spermatozoa (P < 0.01). All the bacterial strains applied affected sperm plasma membrane architecture measured by M540 test (P < 0.01). Moreover, the presence of E. coli or B. ureolyticus was connected with significant decrease in both the number of cells with high mitochondrial transmembrane potential (ΔΨm) and the cells with normal oxidoreductive function of mitochondria (P < 0.05 as compared to untreated cells). To conclude, the contact of bacteria with ejaculated spermatozoa can be a reason for severe injury of sperm membrane stability and mitochondrial activity with potential consequences for male fertility.

Topics: Adult; Bacteroides Infections; Benzimidazoles; Carbocyanines; Cell Membrane; Cell Survival; Escherichia coli Infections; Humans; Infertility, Male; Male; Mitochondria; Pyrimidinones; Spermatozoa; Staphylococcal Infections; Staphylococcus haemolyticus

To establish a high-sensitivity, high-specificity and low-cost hydrogel chip platform for the clinical screening of Y chromosome microdeletions.. Site-specific extended primers with a common sequence at the 5' end were used for hybridizing with the target. The Cy5-dUTP was incorporated into the products by primer extension, and the products were labeled with fluorescence. Then the extended products were added to the chip for hybridizing with acrylamide-modified common probes immobilized on the chip. After removal of the free Cy5-dUTP by electrophoresis, the signals were obtained by fluorescence scanning. And the detecting conditions of this method were optimized.. SY254 of 9 samples was successfully detected with the hydrogel chip. The results showed that 3 were normal and the other 6 with microdeletions (1 female sample as a negative control), which coincided with the results of conventional multiplex PCR-electrophoresis.. The hydrogel chip platform we established has provided a new technique for the detection of Y chromosome microdeletions, and is beneficial to the diagnosis and treatment of male infertility.

Topics: Carbocyanines; Chromosome Deletion; Chromosomes, Human, Y; Deoxyuracil Nucleotides; Humans; Hydrogels; Infertility, Male; Male; Oligonucleotide Array Sequence Analysis; Sex Chromosome Aberrations; Sex Chromosome Disorders of Sex Development

Sperm motility evaluation is associated with fertility in IVF programmes. The visual estimation of sperm motility is extremely subjective. Hence, alternative methods are required. Among them, determination of mitochondrial membrane potential (Deltapsi(m)) changes of spermatozoa using potentiometric dyes may be a reliable test to determine sperm quality. However, the use of the potentiometric dyes in sperm samples has not been compared.. We have studied sperm samples from 28 infertile patients enrolled in an IVF programme in flow cytometry after staining of spermatozoa with four commonly used potentiometric dyes. Sperm motility was evaluated visually.. As expected, JC-1 seems to detect specifically Deltapsi(m) changes, CMX-Ros, DiOC(6)(3) and TMRE fluorescence is easily analysed and the latter three fluorochromes are particularly suitable for multiparametric staining. Irrespective of the Deltapsi(m)-dependent fluorochromes used to stain spermatozoa, a positive correlation was found between the percentage of Deltapsi(m)(high) cells and forward motility and also with high fertilization rates after IVF.. The four fluorochromes may be useful for evaluation of sperm samples from infertile patients. The choice of the potentiometric dyes will depend on their fluorescence characteristics in order to use them in combination with other fluorescent markers.

Topics: Adult; Benzimidazoles; Carbocyanines; Cell Survival; Fertilization; Flow Cytometry; Fluorescent Dyes; Humans; Infertility, Male; Male; Membrane Potentials; Mitochondria; Organic Chemicals; Organometallic Compounds; Semen Preservation; Sensitivity and Specificity; Sperm Motility; Spermatozoa; Staining and Labeling

To evaluate the mitochondrial membrane potential (MMP) of spermatozoa and its correlation with semen parameters and production of reactive oxygen species (ROS) in infertile men and healthy donors.. Controlled prospective study.. Male infertility clinic, Glickman Urological Institute, The Cleveland Clinic Foundation, Cleveland, Ohio.. Nineteen infertile men and 7 healthy volunteers.. Standard semen analysis, assessment of MMP and ROS production in spermatozoa. The MMP was assessed by flow cytometry using the probe carbocyanine DiOC(6)(3) and ROS was measured with chemiluminescence assay using luminol.. The results of MMP are reported as the median interquartile range (IQR) number of cells counted in different areas of fluorescence. Results of ROS measurement are expressed as x10(6) counted photons per minute per 20 million sperm (cpm).. The patients with abnormal semen parameters had a significantly lower MMP [1337.7 (1066.38, 1879.2)], and higher ROS [1.12 (0.26, 3.86)] than the donors [MMP: 2482.9 (2162.5, 3520.6)] and [ROS: 0.10 (0.01, 0.14)]. The MMP was positively correlated with sperm concentration (r = 0.62) and negatively correlated with the ROS produced (r = -0.45).. Measuring MMP in spermatozoa provides useful information about a man's fertility potential. Increased ROS production by spermatozoa is associated with a decreased MMP.

Topics: Carbocyanines; Flow Cytometry; Fluorescent Dyes; Humans; Infertility, Male; Luminescent Measurements; Luminol; Male; Membrane Potentials; Mitochondria; Oxidative Stress; Prospective Studies; Reactive Oxygen Species; Sperm Count; Sperm Motility; Spermatozoa