IFN-gamma alters the human sperm membrane permeability to Ca(2+).

Inflammation in the male genitourinary tract has been associated with the release of pro-inflammatory cytokines such as interferon-gamma (IFN-γ) and elevated reactive oxygen species, which affects spermatozoa capacitation, motility, and the acrosome reaction, along with functions regulated by the concentration of cytoplasmic Ca(2+) ([Ca(2+)]cyto). Though Ca(2+) signaling is of particular significance in sperm, the effect of IFN-γ intracellular calcium on these cells is still unknown. The present study evaluated the effect of IFN-γ on the [Ca(2+)]cyto and Ca(2+) permeability on human sperm. A cell suspension loaded with fura-2 was incubated with or without IFN-γ (from 0 to 2000 pg/ml) for 0, 30, 60, and 120 minutes, and the [Ca(2+)]cyto was measured. The permeability to Ca(2+) was evaluated by the change of the intracellular concentration following an extracellular Ca(2+) pulse. IFN-γ at low concentrations (≤ 500 pg/ml) did not affect the [Ca(2+)]cyto and Ca(2+) permeability of sperm. At a high concentration (2000 pg/ml), IFN-γ did not alter the [Ca(2+)](cyto), but significantly decreased the magnitude and velocity of Ca(2+) entry into the cell. This effect was dependent on incubation time and IFN-γ concentration. This alteration induced by IFN-γ was prevented by the simultaneous incubation of sperm with the antioxidant butylhydroxytoluene (BHT). In conclusion, in vitro, IFN-γ modifies Ca(2+) sperm membrane permeability, probably via lipid peroxidation. IFN-γ in high concentration, as observed in inflammation/infection, can affect [Ca(2+)](cyto) regulation and alter sperm fertilizing capacity.

Effect of tumor necrosis factor-α on the intracellular Ca(2+) homeostasis in human sperm.

PROBLEM: Inflammation and genital infections promote the increase in leukocytes, pro-inflammatory cytokines, and oxygen reactive species, impairing sperm functions such as motility, capacitation, and acrosome reaction. All these functions are primarily regulated by cytoplasmic concentration of Ca(2+) ([Ca(2+) ]cyto ). This study evaluated the effect of tumor necrosis factor (TNF)-α on the [Ca(2+) ]cyto and its regulation in human sperm. METHOD OF STUDY: Sperm loaded with fura-2 were incubated with or without TNF-α (0-500 pg/mL) from 0 to 120 min. After incubation, the basal [Ca(2+) ]cyto and membrane permeability to Ca(2+) were evaluated by spectrofluorometry, before and after Ca(2+) addition to the extracellular medium. RESULTS: Without TNF-α, the addition of Ca(2+) promotes an transitory increase in [Ca(2+) ]cyto in the spermatozoa, that returns in a few minutes to a basal level, indicating calcium regulation activation. TNF-α decreases the Ca(2+) permeation and increases the basal level of [Ca(2+) ]cyto after a Ca(2+) pulse (P < 0.04); affecting calcium regulation in a way that is time and concentration dependent. TNF-α effect was partially prevented by the addition of an antioxidant (butylated hydroxytoluene) (P < 0.03). CONCLUSION: Tumor necrosis factor-α decreases membrane permeability to Ca(2+) and affects Ca(2+) regulation in sperm cells in vitro, probably via lipid peroxidation, which may explain the decrease in sperm fertilizing capacity during inflammatory and infectious processes.

Population genetic structure of the dengue mosquito Aedes aegypti in Venezuela.

The mosquito Aedes aegypti is the main vector of dengue in Venezuela. The genetic structure of this vector was investigated in 24 samples collected from eight geographic regions separated by up to 1160 km. We examined the distribution of a 359-basepair region of the NADH dehydrogenase subunit 4 mitochondrial gene among 1144 Ae. aegypti from eight collections. This gene was amplified by the polymerase chain reaction and tested for variation using single strand conformation polymorphism analysis. Seven haplotypes were detected throughout Venezuela and these were sorted into two clades. Significant differentiation was detected among collections and these were genetically isolated by distance.

Population genetic structure of the dengue mosquito Aedes aegypti in Venezuela

The mosquito Aedes aegypti is the main vector of dengue in Venezuela. The genetic structure of this vector was investigated in 24 samples collected from eight geographic regions separated by up to 1160 km. We examined the distribution of a 359-basepair region of the NADH dehydrogenase subunit 4 mitochondrial gene among 1144 Ae. aegypti from eight collections. This gene was amplified by the polymerase chain reaction and tested for variation using single strand conformation polymorphism analysis. Seven haplotypes were detected throughout Venezuela and these were sorted into two clades. Significant differentiation was detected among collections and these were genetically isolated by distance.