Metformin Improves Quality of Post-Thaw Canine Semen

Abstract

Sperm cryopreservation is an assisted reproductive technique routinely used in canine species for genetic conservation. However, during cryopreservation, the DNA damages are still elevated, limiting the fertilization rate. The present study was conducted to evaluate whether supplementation of canine semen extender with a molecule limiting the metabolic activities can improve the quality of frozen-thawed canine spermatozoa. We used metformin, known to limit the mitochondrial respiratory and limit the oxidative stress. Before and during the freezing procedure, metformin (50µM and 500µM) has been added to the extender. After thawing, sperm exposed to metformin conserved the same viability without alteration in the membrane integrity or acrosome reaction. Interestingly, 50µM metformin improved the sperm motility in comparison to the control, subsequently increasing mitochondrial activity and NAD+ content. In addition, the oxidative stress level was reduced in sperm treated with metformin improving the sperm quality as measured by a different molecular marker. In conclusion, we have shown that metformin is able to improve the quality of frozen-thawed dog semen when it is used during the cryopreservative procedure.

Antioxidant effects of supplementation of 3,4‐dihydroxyphenyl glycol on sperm parameters and oxidative markers following cryopreservation in canine semen

“…We also concluded that antioxidants could enhance the plasma membrane integrity of the spermatozoa that was is in agreement with the results obtained by Munsi et al., (2007) and Patel et al., (2015). Grandhaye et al., (2020) showed that the use of metformin in canine sperm has decreased the oxidative stress and limited the percentage of spermatozoa with damaged DNA. The positive relationship between hypo‐osmotic swelling test (HOST) positive sperm and variables like motility, progressive motility, sperm concentration and total intact acrosome was concluded by Srivastava and Kumar (2006).…”

Conditioned Medium from Canine Amniotic Membrane-Derived Mesenchymal Stem Cells Improved Dog Sperm Post-Thaw Quality-Related Parameters

“…Although Tris-egg yolk buffer is the most commonly used diluent for mammalian sperm, many studies show that the supplementation of anti-apoptotic factors, anti-oxidants, post-thaw enhancing chemicals, or novel cryo-protective agents is required to get good post-thaw results [ 1 , 5 , 17 , 18 ]. Thus, various molecules such as metformin [ 19 ], cholesterol [ 20 ], or α-tocopherol [ 21 ] have been successfully used in canine sperm cryopreservation to reduce oxidative stress, DNA damage [ 19 , 20 ], improve motility [ 21 ], and protect plasma membrane and acrosome integrity [ 20 ].…”

The physiology of male reproduction: Impact of drugs and their abuse on male fertility

“…McPherson and Lane (2020) also observed that metformin treatment restored sperm function and foetal growth without requiring weight loss in high‐fat diet obese male mice. Furthermore, Grandhaye et al, (2020) provided evidence to suggest that metformin improved the quality of canine cryopreserved semen via AMPK pathway.…”