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. 2023 Feb 16:10:1093274.
doi: 10.3389/fvets.2023.1093274. eCollection 2023.

Glutathione and selenium nanoparticles have a synergistic protective effect during cryopreservation of bull semen

Shubin Li  1 , Jingyu Ren  2 , Wenqi Zhang  1 , Biao Wang  3 , Yuzhen Ma  4 , Liya Su  1 , Yanfeng Dai  2 , Gang Liu  1
Affiliations

Glutathione and selenium nanoparticles have a synergistic protective effect during cryopreservation of bull semen

Shubin Li et al. Front Vet Sci. .

Abstract

Introduction: In the present study, the synergistic protective effect of co-supplementation of glutathione (GSH) with selenium nanoparticles (SeNPs) on the cryopreservation efficiency of bull semen was analyzed.

Methods: After collection, the ejaculates of Holstein bulls were subsequently diluted with a Tris extender buffer supplemented with different concentrations of SeNPs (0, 1, 2, and 4 μg/ml), followed by semen equilibration at 4°C and assessment of sperm viability and motility. Subsequently, the ejaculates of Holstein bulls were pooled, split into four equal groups, and diluted with a Tris extender buffer supplemented with basic extender (negative control group, NC group), 2 μg/ml SeNPs (SeNPs group), 4 mM GSH (GSH group), and 4 mM GSH plus 2 μg/ml SeNPs (GSH + SeNPs group). After cryopreservation, motility, viability, mitochondrial activity, plasma membrane integrity, acrosome integrity, concentration of malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT), and ability of frozen-thawed sperm cells to support in vitro embryonic development were evaluated.

Results and discussion: No side effect of SeNPs concentrations applied in the current study on the motility and viability of equilibrated bull spermatozoa was found. Meanwhile, supplementation of SeNPs significantly promoted the motility and viability of equilibrated bull spermatozoa. Furthermore, the co-supplementation of GSH with SeNPs effectively protected bull spermatozoa from cryoinjury as expressed by promoting semen motility, viability, mitochondrial activity, plasma membrane integrity, and acrosome integrity. Finally, the enhanced antioxidant capacity and embryonic development potential in the frozen-thawed bull spermatozoa cryopreserved by co-supplementation of GSH with SeNPs further confirmed the synergistic protective effect of co-supplementation of GSH with SeNPs on the cryopreservation of bull semen.

Keywords: GSH; SeNPs; bull; oxidative stress; semen cryopreservation.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Effect of different concentrations of SeNPs supplementation on motility and viability of equilibrated bull spermatozoa. (A) Motility of equilibrated bull spermatozoa. (B) Viability of equilibrated bull spermatozoa. Labeling with a different lower-case letter in each column indicates significant differences among groups (n = 3, P < 0.05).
Figure 2
Figure 2
Effect of co-supplementation of GSH and SeNPs on motility and viability of cryopreserved bull spermatozoa. (A) Progressive motility of cryopreserved bull spermatozoa. (B) Viability of cryopreserved bull spermatozoa. Labeling with a different lowercase letter in each column indicates significant differences among groups (n = 6, P < 0.05).
Figure 3
Figure 3
Effect of co-supplementation of GSH and SeNPs on mitochondrial activity, acrosome integrity, and plasma membrane integrity of cryopreserved bull spermatozoa. (A) Mitochondrial activity of cryopreserved bull spermatozoa. (B) Acrosome integrity of cryopreserved bull spermatozoa. (C) Plasma membrane integrity of cryopreserved bull spermatozoa. Labeling with a different lower-case letter in each column indicates significant differences among groups (n = 3, P < 0.05).
Figure 4
Figure 4
Effect of GSH and SeNPs co-supplementation on antioxidant potentials of cryopreserved bull spermatozoa. (A) MDA levels of cryopreserved bull spermatozoa. (B) SOD levels of cryopreserved bull spermatozoa. (C) CAT levels of cryopreserved bull spermatozoa. Labeling with a different lower-case letter in each column indicates significant differences among groups (n = 3, P < 0.05).
Figure 5
Figure 5
Effect of GSH and SeNPs co-supplementation on cleavage and blastocyst development of cryopreserved bull spermatozoa. (A) Cleavage rates of cryopreserved bull spermatozoa. (B) Percentage of blastocysts/cleaved 2-cell embryos. Labeling with a different lower-case letter in each column indicates significant differences among groups (n = 3, P < 0.05).
Figure 6
Figure 6
Effect of GSH and SeNPs co-supplementation on blastocyst development of cryopreserved bull spermatozoa. (A) Hoechst 33342 staining of the bovine blastocyst. Scale bar = 20 μm. (B) Cell number/blastocyst of each group. Labeling with a different lowercase letter in each column indicates significant differences among groups (n = 20, P < 0.05).
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