Unveiling the impact of environmental microplastics on mussel spermatozoa: First evidence of prothymosin-α detection in invertebrate's male gametes.

Mytilus galloprovincialis mussels, like many other marine invertebrates, employ external fertilization as a mating strategy, exposing their gametes to various contaminants upon release into seawater. Environmental microplastics (EMP) are prevalent marine pollutants that pose a significant threat to aquatic biota. In this regard, our study aimed to investigate the potential effects of exposing mussels' male gametes to increasing concentrations of EMP (1, 10, 50, and 100 μg/l) collected from a Mediterranean sandy beach. We focused on assessing gamete quality by analysing physiological parameters such as viability, mitochondrial membrane potential, oxidative status, and motility. Additionally, we evaluated DNA integrity and activation of apoptosis. Furthermore, our study aimed to detect the presence of the prothymosin-α (PTMA) protein, which has never been previously investigated in invertebrate spermatozoa. Our data revealed that exposure of mussel spermatozoa to EMPs altered their oxidative status and mitochondrial membrane potential, induced a decrease in motility, DNA integrity, and an increased apoptotic occurrence, leading to a decline in overall viability. The localization of PTMA into the head and flagellum of spermatozoa further supported its presence and susceptibility to the effects of microplastics. These findings raise concerns about the reproductive capacity of mussels under environmental microplastic pollution and highlight potential long-term threats to population sustainability.