Here we report the industrial pollution effects due to cadmium on the reproductive health of Mytilus galloprovincialis. Mussels were removed from the biofouling of a Conatex panel after one year exposition at a polluted site near a disposal metallurgical factory. A high cadmium bioaccumulation was observed in the testis of mussels housed at the polluted site, with respect to a control site, as determined by inductively coupled plasma-mass spectrometry, along with a 10 fold increase in metallothionein 20 kDa gene (mt20) expression levels determined by qPCR. Furthermore, mussels transferred into laboratory tanks from the reference site, and exposed to 1.5, 5 and 10 mu M CdCl2, revealed a 1.7, 3.2 and 4.5 fold expression increase in the testis mt20, respectively, and a positive correlation with cadmium bioaccumulation was found. To evaluate a potential detrimental risk of such alterations on spermatozoa, we carried out electrophoretic analyses on their protamine-like proteins. As determined by AU-PAGE, after 1.5 mu M CdCl2 exposure, protamine-like proteins also display major alterations with respect to those obtained after 5 and 10 mu M CdCl2 exposure. All protamine-like proteins isolated from the polluted biofouling were in an aggregated form and displayed the same reduced DNA binding affinity of the protamine-like proteins obtained after 1.5 mu M CdCl2 as demonstrated EMSA with sperm genomic DNA. Our results contribute to the studies concerning cadmium induced testis alterations and highlight protamine-like proteins' analysis as an emerging biotechnique for cadmium impact assessment on Mytilus galloprovincialis, for the sensitivity of the in vivo and in vitro changes of protamine-like proteins' state and their DNA binding affinity.

Protamine-like proteins' analysis as an emerging biotechnique for cadmium impact assessment on male mollusk Mytilus galloprovincialis (Lamarck 1819) / Piscopo, Marina; Trifuoggi, Marco; Notariale, Rosaria; Labar, Sofiane; Troisi, Jacopo; Giarra, Antonella; Rabbito, Dea; Puoti, Raffaela; de Benedictis, Domenico; Brundo, Maria Violetta; Basile, Adriana; Good, Katrina V.; Ausió, Juan; Guerriero, Giulia. - In: ACTA BIOCHIMICA POLONICA. - ISSN 0001-527X. - 65:2(2018), pp. 259-267. [10.18388/abp.2017_2533]

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

Piscopo, Marina
;
Trifuoggi, Marco;Giarra, Antonella;Rabbito, Dea;Basile, Adriana;Guerriero, Giulia
2018

Abstract

Here we report the industrial pollution effects due to cadmium on the reproductive health of Mytilus galloprovincialis. Mussels were removed from the biofouling of a Conatex panel after one year exposition at a polluted site near a disposal metallurgical factory. A high cadmium bioaccumulation was observed in the testis of mussels housed at the polluted site, with respect to a control site, as determined by inductively coupled plasma-mass spectrometry, along with a 10 fold increase in metallothionein 20 kDa gene (mt20) expression levels determined by qPCR. Furthermore, mussels transferred into laboratory tanks from the reference site, and exposed to 1.5, 5 and 10 mu M CdCl2, revealed a 1.7, 3.2 and 4.5 fold expression increase in the testis mt20, respectively, and a positive correlation with cadmium bioaccumulation was found. To evaluate a potential detrimental risk of such alterations on spermatozoa, we carried out electrophoretic analyses on their protamine-like proteins. As determined by AU-PAGE, after 1.5 mu M CdCl2 exposure, protamine-like proteins also display major alterations with respect to those obtained after 5 and 10 mu M CdCl2 exposure. All protamine-like proteins isolated from the polluted biofouling were in an aggregated form and displayed the same reduced DNA binding affinity of the protamine-like proteins obtained after 1.5 mu M CdCl2 as demonstrated EMSA with sperm genomic DNA. Our results contribute to the studies concerning cadmium induced testis alterations and highlight protamine-like proteins' analysis as an emerging biotechnique for cadmium impact assessment on Mytilus galloprovincialis, for the sensitivity of the in vivo and in vitro changes of protamine-like proteins' state and their DNA binding affinity.
2018
Protamine-like proteins' analysis as an emerging biotechnique for cadmium impact assessment on male mollusk Mytilus galloprovincialis (Lamarck 1819) / Piscopo, Marina; Trifuoggi, Marco; Notariale, Rosaria; Labar, Sofiane; Troisi, Jacopo; Giarra, Antonella; Rabbito, Dea; Puoti, Raffaela; de Benedictis, Domenico; Brundo, Maria Violetta; Basile, Adriana; Good, Katrina V.; Ausió, Juan; Guerriero, Giulia. - In: ACTA BIOCHIMICA POLONICA. - ISSN 0001-527X. - 65:2(2018), pp. 259-267. [10.18388/abp.2017_2533]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11588/720112
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