Treatment of sperm with extracellular adenosine 5'-triphosphate improves the in vitro fertility rate of inbred and genetically modified mice with low fertility

Abstract

In vitro fertilization (IVF) is one of the most important techniques used for assisted reproduction in mouse colony management. As with natural mating, where mice have varying fertility indices, fertility rates of genetically modified (GM) [transgenic (Tg), knock out (KO) and congenic (Cg)] mice are influenced by their genetic background. Lines of GM mice that have poor fertility have a concomitant poor IVF outcome. Treatment of mouse sperm with extracellular adenosine 5’-triphosphate (ATPe) enhanced in vitro fertilization rates in outbred and hybrid mice. The objective of this study was to analyze the effects of using extracellular adenosine 5’-triphosphate-treated sperm for IVF of inbred wild type, and genetically modified mouse lines, for which standard IVF did not work well. The IVF was performed using the GM mice on C57BL/10SnJ, C57BL/6J, BALB/cJ and NFS/N background strains and wild type (WT) mice such as C57BL/6N, BALB/cAnN, and B6129SF1 strains. Oocytes from superovulated females were fertilized in vitro with sperm from the same background strain, and either treated or not treated with ATPe. The ATPe treatment enhanced IVF outcome in most of the GM and some WT strains, as indicated by the percentage of embryos that progressed to the two-cell stage. There was no marked difference between ATPe treated and control groups for the development rate of two-cell embryos to blastocysts in culture, or in the number of pups born after transfer of two-cell embryos into recipient females. The observed improvement of the IVF results following ATPe treatment of transgenic and KO mouse sperm were a potential solution for improving the outcome of assisted reproduction techniques used for rederivation or for gamete banking.

Purinergic signalling in the reproductive system in health and disease

“…Treatment of sperm with ATP used for IVF is now being explored. For example, a recent paper showed, with IVF experiments in mice, that treatment of sperm with ATP improved the in vitro fertility rate of genetically modified (transgenic and knockout) mice with low fertility [411].…”

Ecto-nucleotidases distribution in human cyclic and postmenopausic endometrium

“…P2X and P2Y nucleotide receptors have been identified in the female reproductive tract [7][8][9], with changes in the expression along the cycle [10] and during implantation [11] and pregnancy [8]. Furthermore, extracellular ATP treatment of sperm improves its fertilizing capability [12][13][14], thus potentially improving the outcome of assisted reproduction techniques.…”

In Vitro Fertilization Activators for Future

“…On the other hand, IVF efficiency using the sperm of aged BALB/cA mice (>48 weeks of age) is low, albeit fluctuated between mice [27]. However, we found that the licorice crude extract improved the fertilizing ability of BALB/cA mouse sperm in vitro as indicated in Figure 2 and that the fertilized eggs developed normally [28].…”