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. 2023 Jan 4;24(2):935.
doi: 10.3390/ijms24020935.

F4-Neuroprostane Effects on Human Sperm

Elena Moretti  1 , Cinzia Signorini  1 , Daria Noto  1 , Roberta Corsaro  1 , Lucia Micheli  2 , Thierry Durand  3 , Camille Oger  3 , Jean Marie Galano  3 , Giulia Collodel  1
Affiliations

F4-Neuroprostane Effects on Human Sperm

Elena Moretti et al. Int J Mol Sci. .

Abstract

Swim-up selected human sperm were incubated with 7 ng F4-neuroprostanes (F4-NeuroPs) for 2 and 4 h. Sperm motility and membrane mitochondrial potential (MMP) were evaluated. The percentage of reacted acrosome was assessed by pisum sativum agglutinin (PSA). Chromatin integrity was detected using the acridine orange (AO) assay and localization of the ryanodine receptor was performed by immunofluorescence analysis. Sperm progressive motility (p = 0.02) and the percentage of sperm showing a strong MMP signal (p = 0.012) significantly increased after 2 h F4-NeuroP incubation compared to control samples. The AO assay did not show differences in the percentage of sperm with dsDNA between treated or control samples. Meanwhile, a significantly higher number of sperm with reacted acrosomes was highlighted by PSA localization after 4 h F4-NeuroP incubation. Finally, using an anti-ryanodine antibody, the immunofluorescence signal was differentially distributed at 2 and 4 h: a strong signal was evident in the midpiece and postacrosomal sheath (70% of sperm) at 2 h, whereas a dotted one appeared at 4 h (53% of sperm). A defined concentration of F4-NeuroPs in seminal fluid may induce sperm capacitation via channel ions present in sperm cells, representing an aid during in vitro sperm preparation that may increase the positive outcome of assisted fertilization.

Keywords: F4-neuroprostanes; acrosome reaction; capacitation; human sperm; in vitro study; ryanodine receptor; sperm motility.

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Conflict of interest statement

All authors declare that they do not have any conflict of interest that could inappropriately influence this manuscript.

Figures

Figure 1
Figure 1
Evaluation of the acrosome status with TRITC–pisum sativum agglutinin (TRITC–PSA). Intact acrosome (a), reacted or partially reacted acrosome (b,c) with no fluorescence or only fluorescence of the equatorial segment and loss of acrosomal membranes. Bars: 6 μM.
Figure 2
Figure 2
UV micrographs of swim-up selected sperm treated with anti-ryanodine antibody. The percentage of localization differed in control samples or those incubated with F4-NeuroPs for 2 or 4 h. In the controls, a high percentage of sperm showed diffuse and fair labeling (a) or a spot localized in the centriolar region (b). Sperm incubated for 2 h with F4-NeuroPs showed strong signals in the midpiece and postacrosomal sheath (c,d). After 4 h F4-NeuroP incubation, the labeling appeared dotted along the total tail (e,f). Nuclei (blue) were stained with DAPI. Figure 2 without DAPI was included in Supplementary Materials. Bars: 6 μM.
Figure 3
Figure 3
Representation of chemical structures of 4- and 10-F4t-NeuroP isomers.
See this image and copyright information in PMC

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