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. 2022 Sep 3;19(17):11023.
doi: 10.3390/ijerph191711023.

Molecular Alterations and Severe Abnormalities in Spermatozoa of Young Men Living in the "Valley of Sacco River" (Latium, Italy): A Preliminary Study

Pasquale Perrone  1   2 , Gennaro Lettieri  1 , Carmela Marinaro  1 , Valentina Longo  3 , Simonetta Capone  3 , Angiola Forleo  3 , Sebastiana Pappalardo  4 , Luigi Montano  5   6 , Marina Piscopo  1
Affiliations

Molecular Alterations and Severe Abnormalities in Spermatozoa of Young Men Living in the "Valley of Sacco River" (Latium, Italy): A Preliminary Study

Pasquale Perrone et al. Int J Environ Res Public Health. .

Abstract

The Valley of Sacco River (VSR) (Latium, Italy) is an area with large-scale industrial chemical production that has led over time to significant contamination of soil and groundwater with various industrial pollutants, such as organic pesticides, dioxins, organic solvents, heavy metals, and particularly, volatile organic compounds (VOCs). In the present study, we investigated the potential impact of VOCs on the spermatozoa of healthy young males living in the VSR, given the prevalent presence of several VOCs in the semen of these individuals. To accomplish this, spermiograms were conducted followed by molecular analyses to assess the content of sperm nuclear basic proteins (SNBPs) in addition to the protamine-histone ratio and DNA binding of these proteins. We found drastic alterations in the spermatozoa of these young males living in the VSR. Alterations were seen in sperm morphology, sperm motility, sperm count, and protamine/histone ratios, and included significant reductions in SNBP-DNA binding capacity. Our results provide preliminary indications of a possible correlation between the observed alterations and the presence of specific VOCs.

Keywords: human protamines; human spermatozoa; male fertility; protein–DNA binding; reproduction; sperm nuclear basic proteins; spermatozoa morphology; volatile organic compounds.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Map of the geographical areas selected for the recruitment. The green circle indicates the Valley of Sele River in the Campania region; the red circle indicates the Valley of Sacco River (VSR) area in the Lazio region. The green and red circles indicate the geographical position of the two areas in the respective regions.
Figure 2
Figure 2
Frequency of VOCs which were detected in semen from the VSR group.
Figure 3
Figure 3
Anthropometric data: (a) age; (b) weight; (c) height and (d) BMI of control group (green) and VSR (red) individuals. BMI: Body Mass Index.
Figure 4
Figure 4
Statistical analysis of sperm concentration (a) and total motility (b) of the two groups. In green, control group, in red, VSR group. **: p ≤ 0.01; ****: p ≤ 0.0001.
Figure 5
Figure 5
Diff-quick staining of VSR group’s spermatozoa. (a): control group; (b–i): VSR group.
Figure 6
Figure 6
SNBP content as characterized by AU-PAGE displayed three different expression patterns. A normal canonical protamine/histone ratio (CP/Hr) was seen mainly in control group samples ((a), lane 10; (b), lane 1), while VSR samples more commonly displayed a histone-only ratio ((a), lanes 1–9) or a non-canonical protamine/histone ratio ((b), lanes 2–10). (c) Percentage distribution of protamines/histones ratios found in spermatozoa. In green, control group, in red, VSR group. CP/Hr: canonical protamine/histones ratio; nCP/Hr: non-canonical protamine/histones ratio; only-H: only histones.
Figure 7
Figure 7
DNA-binding ability of SNBPs obtained from control group (a) and VSR group (b,c) analyzed by electrophoretic mobility shift assay (EMSA) on 1% agarose gel. Bands on gel representing the state of pGEM3 plasmid DNA incubated in a ratio w/w with increasing amount of SNBPs from samples. VSR: Valley of Sacco River; control: Valley of Sele River.
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