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  • New Insight on the In Vitro Effects of Melatonin in Preserving Human Sperm Quality.

New Insight on the In Vitro Effects of Melatonin in Preserving Human Sperm Quality.

International journal of molecular sciences (2022-05-15)
Sergio Minucci, Massimo Venditti
ABSTRACT

Spermatozoa (SPZ) are sensitive to stressful conditions, particularly oxidative stress, which alters their quality; thus, the use of protective molecules as an antioxidant is encouraged. Herein, we used melatonin (MLT) to investigate its in vitro effects on human sperm parameters under conditions of oxidative stress induced by cadmium (Cd). Fifteen human semen samples were divided into control, Cd-treated, MLT-treated, and Cd+MLT-treated groups and analyzed after 30 min, 6 h, and 24 h of exposure. Results showed a time-dependent decrease in SPZ motility, DNA integrity, and increased apoptosis induced by oxidative stress, and these effects were counteracted by MLT co-treatment. Based on these data, we further explored additional parameters just at 24 h. The induced oxidative stress, highlighted by the increased lipid peroxidation, reduced the percentage of SPZ able to undertake acrosome reaction and altered the levels and localization of some protein markers of motility (PREP, RSPH6A), morphology (DAAM1), and acrosome membrane (PTMA, IAM38); all these effects were counteracted by MLT co-treatment. Interestingly, MLT alone was able to ameliorate motility at 30 min of incubation compared to the control, while at 24 h, it prevented the physiological alteration in terms of motility, DNA integrity, and apoptosis. Collectively, the data encourage MLT use as an integrative molecule to ameliorate human gamete quality when compromised by stressful conditions.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, crystallized, ≥98.0% (GE)
Sigma-Aldrich
Goat Anti-Mouse IgG & IgM Antibody, HRP conjugate, 0.8 mg/mL, Chemicon®
Sigma-Aldrich
Anti-RSPH6A antibody produced in rabbit, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution
Sigma-Aldrich
Anti-Mouse IgG (H+L), CF 568 antibody produced in goat, ~2 mg/mL, affinity isolated antibody
Sigma-Aldrich
Goat Anti-Rabbit IgG Antibody, (H+L) HRP conjugate, 1 mg/mL, Chemicon®