Sperm viability assessment in marine invertebrates by fluorescent staining and spectrofluorimetry: A promising tool for assessing marine pollution impact

Gallo, Alessandra; Boni, Raffaele; Tosti, Elisabetta

doi:10.1016/j.ecoenv.2017.07.069

Cited by 15 publications

(9 citation statements)

References 38 publications

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“…These effects were evaluated together with standard computer-assisted sperm kinetics, by using a simple, accurate, rapid, and sensitive methodology, which combines fluorescent probes with spectrofluorimetric analysis for the assessment of ΔΨM, lipid peroxidation, ROS production and pH i in mammalian (human and bovine), as well as in the marine invertebrate ( C. robusta ) spermatozoa. In human sperm, this methodology is a novelty whereas in bovine and ascidian sperm this methodological approach have been partially tested in recent studies 26 – 28 . Beneficial effects of NAC on semen parameters including sperm motility have been already described in several mammalian species, such as bovine 29 , human 30 as well as in the rooster 31 .…”

Section: Discussionmentioning

confidence: 99%

Metabolic enhancers supporting 1-carbon cycle affect sperm functionality: an in vitro comparative study

Gallo

Ménézo²,

Dale³

et al. 2018

Sci Rep

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The sperm plasma membrane is a sensitive target to oxidative stress. The most representative reactive oxygen species (ROS) scavengers in the genital tract, hypotaurine and glutathione, require, for their synthesis, cysteine whose availability is associated with the 1-carbon cycle (1-CC). Human, bovine and ascidian spermatozoa were incubated with compounds supporting the 1-CC (Vitamin B6, Methylcobalamin, 5 Methyl Tetrahydrofolate, Zinc Bisglycinate and N-acetyl-cysteine) (TRT) and compared to the effects induced solely by N-acetyl-cysteine (NAC). In control groups (CNTRL), spermatozoa were incubated with medium alone. After 90 and 180 minutes of incubation, the mitochondrial membrane potential (ΔΨM) in TRT and NAC was significantly (P < 0.01) higher than in CNTRL. At H2DCFDA evaluation, ROS production differed between species whereas, at 2-OH Ethidium, it significantly decreased in bovine TRT group. Intracellular pH (pHi) did not significantly vary in relation to treatment. In ascidian spermatozoa, the NAC supplementation decreased external pH, which in turn brought to a pHi lowering. Buffering seawater with NaHCO3 reversed the beneficial effects of N-acetyl-cysteine supplementation. In conclusion, both fully supporting the 1-CC and treatment with N-acetyl-cysteine alone improved kinetics, ΔΨM and ROS production in mammalian sperm demonstrating for the first time the direct in vitro effects of these compounds on sperm functionality.

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Section: Discussionmentioning

confidence: 99%

Metabolic enhancers supporting 1-carbon cycle affect sperm functionality: an in vitro comparative study

Gallo

Ménézo²,

Dale³

et al. 2018

Sci Rep

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“…Contrastingly, PI permeates only spermatozoa that lost membrane integrity. Briefly, according to Gallo et al (2018), aliquots of spermatozoa diluted at 5 × 10 6 cell/mL were added to final testing suspensions of CuO NPs and incubated for 1 h at 18 °C. After incubation, spermatozoa were stained by adding SYBR 14 (final concentration 100 nM) and incubated in the dark for 15 min.…”

Section: Sperm Viabilitymentioning

confidence: 99%

Cytotoxicity and genotoxicity of CuO nanoparticles in sea urchin spermatozoa through oxidative stress

Gallo

Manfra

Boni

et al. 2018

Environment International

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“…In the past, many marine organisms have been proposed as bioindicators of environmental pollution such as echinoderms and molluscs (Bellas, 2007; Nacci et al ., 1986). In particular sea urchin early embryo growth test and sperm cell toxicity test have been used for long time to evaluate toxicity of a closed environment and recently new modified bioassays have been found to be good alternatives to the traditional tests (Manzo, 2004; Matranga & Corsi, 2012; Gallo et al , 2018). At present, there is a need to identify new genotoxic and teratogenic tests to associate with environmental quality monitoring.…”

Section: Discussionmentioning

confidence: 99%

Toxicity of marine pollutants on the ascidian oocyte physiology: an electrophysiological approach

Gallo

2017

Zygote

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In marine animals with external fertilization, gametes are released into seawater where fertilization and embryo development occur. Consequently, pollutants introduced into the marine environment by human activities may affect gametes and embryos. These xenobiotics can alter cell physiology with consequent reduction of fertilization success. Here the adverse effects on the reproductive processes of the marine invertebrate Ciona intestinalis (ascidian) of different xenobiotics: lead, zinc, an organic tin compound and a phenylurea herbicide were evaluated. By using the electrophysiological technique of whole-cell voltage clamping, the effects of these compounds on the mature oocyte plasma membrane electrical properties and the electrical events of fertilization were tested by calculating the concentration that induced 50% normal larval formation (EC50). The results demonstrated that sodium currents in mature oocytes were reduced in a concentration-dependent manner by all tested xenobiotics, with the lowest EC50 value for lead. In contrast, fertilization current frequencies were differently affected by zinc and organic tin compound. Toxicity tests on gametes demonstrated that sperm fertilizing capability and fertilization oocyte competence were not altered by xenobiotics, whereas fertilization was inhibited in zinc solution and underwent a reduction in organic tin compound solution (EC50 value of 1.7 µM). Furthermore, fertilized oocytes resulted in a low percentage of normal larvae with an EC50 value of 0.90 µM. This study shows that reproductive processes of ascidians are highly sensitive to xenobiotics suggesting that they may be considered a reliable biomarker and that ascidians are suitable model organisms to assess marine environmental quality.

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Sperm viability assessment in marine invertebrates by fluorescent staining and spectrofluorimetry: A promising tool for assessing marine pollution impact

Cited by 15 publications

References 38 publications

Metabolic enhancers supporting 1-carbon cycle affect sperm functionality: an in vitro comparative study

Metabolic enhancers supporting 1-carbon cycle affect sperm functionality: an in vitro comparative study

Cytotoxicity and genotoxicity of CuO nanoparticles in sea urchin spermatozoa through oxidative stress

Toxicity of marine pollutants on the ascidian oocyte physiology: an electrophysiological approach

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