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Mitochondrial membrane potential (ΔΨm)The ΔΨm was measured using the ﬂuorescent dye 5,5', 6, 6'- tetra-chloro-1,1', 3, 3'- tetraethylben-zimidazolyl-carbocyanine iodide (JC-1), a cell permeable cationic dye that can accumulate in energized mitochondria based on the highly negative ΔΨm.
Essentially, JC-1 changes its fluorescence from green (monomeric status) to red (multimeric status) when the ΔΨm is high. The stained samples were then analyzed by flow cytometry to determine the sperm with high ΔΨm (red stained) using the appropriate gating parameters.
The motility of 200 spermatozoa was evaluated by CASA.
The sperm motility parameters were evaluated after 3 and 6 h of incubation with 0, 1, 5, or 25 mg l Aroclor 1254.
Ten thousand sperm from each group were used in the evaluation of ΔΨm.
Intracellular ROS production DCFH-DA, an oxidation-sensitive fluorescent probe, was used to analyze ROS generation. The cellular mean fluorescence intensity (MFI) was measured using flow cytometry.
Chemicals Aroclor 1254 was purchased from Santa Cruz Biotechnology Inc. All other chemicals used in the present study were obtained from Sigma-Aldrich (St. Human biological samples Sperm samples with ≥15 × 10 were obtained from healthy donors (n = 12) undergoing routine semen analysis in the Qilu Hospital of Shandong University (China).
Toxic effects of polychlorinated biphenyls (Aroclor 1254) on human sperm motility.
Nevertheless, there remains concern about human exposure due to the growing number of poisoning incidents, such as the inadvertent ingestion of dairy products or seafood contaminated by PCBs.
These toxic compounds also appear to accumulate in organisms and can cause endocrine disruption and reproductive disorders, such as infertility and miscarriage.
One class of persistent organic pollutant, polychlorinated biphenyls (PCBs), was originally manufactured for industrial applications due to its insulating and flame-retardant properties.
Several recent studies on PCBs have found that they possess endocrine disrupting activity.To this aim, a short-term in vitro incubation experiment that better imitated the putative exposure of sperm to Aroclor 1254 (a commercial PCB mixture) in male reproduction tissue was conducted.