Mytilus galloprovincialis (Lamarck, 1819) spermatozoa: hsp70 expression and protamine-like protein property studies

Abstract

In this work, we describe results of the reproductive health monitoring studies in Mytilus galloprovincialis following spermatozoa hsp70 expression and protamine-like protein properties. Mussels control (ctr) were released within cages for 30 days in three different marine sites near Naples (Campania, Italy): Bagnoli south (BAs) and Bagnoli north (BAn), both close to a disposal metallurgical factory and in Capo Miseno (CM). Studies of hsp70 gene expression carried out, by RT-qPCR, in mussel spermatozoa have shown varied expression levels, particularly 5, 13, and 15-fold more than ctr in CM, BAs, and BAn, respectively, indicating highest involvement of stress proteins in spermatozoa of mussels in Bagnoli. In order to evaluate the possible risk on Mytilus galloprovincialis sustainability loss, electrophoretic analyses were performed on protamine-like proteins (PL) of collected spermatozoa. The results showed that CM PL were apparently unaltered with respect to ctr PL, while BAs and BAn PL appeared in part in the form of peptides and in part as bands with low mobility. Further, CM and BAs PL showed, by electrophoretic mobility shift assay, a decrease in DNA binding ability and a change in their DNA binding mode. The results of this investigation show the usefulness of the study of alterations of spermatozoa hsp70 expression and protamine-like protein properties for eco-toxicological evaluation using Mytilus galloprovincialis as a bioindicator.

Protamine-like proteins analyses as emerging biotechnique for cadmium impact assessment on male mollusk Mytilus galloprovincialis (Lamarck 1819)

“…Recently, we have reported results of the reproductive health monitoring studies in Mytilus galloprovincialis following spermatozoa hsp70 expression and PL-proteins' properties. In particular, we found alterations in the PLproteins' electrophoretic pattern and in their DNA binding mode and affinity (Piscopo et al, 2018b). Thus, in this work we studied industrial pollution by using mussels removed from the biofouling of a Conatex panel after one year of exposition when the panel is completely covered only by mussels, at a polluted site close to a disposal metallurgical factory: Ilva in Bagnoli (Naples, Campania -Italy).…”

Temperature changes and marine fish species (Epinephelus coioides and Sparus aurata): Role of oxidative stress biomarkers in toxicological food studies

“…Results indicated that fish showed great response to different temperature limits, that higher HSPs (HSP70 and HSP90) were up-regulated during stable elevatedtemperature level exposure time (Podrabsky and Somero, 2004;Piscopo et al, 2018;Somero, 2010).…”

Protamine-like proteins have bactericidal activity. The first evidence in Mytilus galloprovincialis

“…This is because marine invertebrates, living within very difficult, competitive, and hostile surroundings represent a great reservoir for compounds with improved bactericidal activity. Mytilus galloprovincialis is an excellent bioindicator and bioaccumulator of marine pollution (Piscopo et al, 2016) and as we have recently shown, it has altered levels of hsp70 in spermatozoa, gonadal and gill tissues (Piscopo et al, 2018c; and demonstrates changes in sperm protaminelike protein properties (Piscopo et al, 2018b, Piscopo et al, 2018c, as a response to environmental stress/geographical site of this organism. The first work concerning bactericidal activity identified in mollusks was relative to the mucus of the giant snail Achatina fulica (Kubota et al, 1985).…”

“…Spermatozoa collection from M. galloprovincialis mussels was performed as previously described (Piscopo et al, 2018b). The crude extract containing the three PL-proteins from M. galloprovincialis spermatozoa was obtained by extraction with 5% PCA as previously described (Piscopo et al, 2018c), starting from a pool of spermatozoa collected from 20 Mytilus galloprovincialis specimens. M. galloprovincialis PL-II and PL-III were fractionated by ionic exchange resin cellulose Whatman CM52 as previously described (Vassalli et al, 2015).…”