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Ruthenium red (RR) inhibits calcium (Ca 2+ ) entry from the cytoplasm to the mitochondria, and is involved in maintenance of Ca 2+ homeostasis in mammalian cells. Ca 2+ homeostasis is very important for further embryonic development of fertilized oocytes. However, the effect of RR on mitochondria-Ca 2+ (mito-Ca 2+ ) levels during in vitro fertilization (IVF) on subsequent blastocyst developmental capacity in porcine is unclear. The present study explored the regulation of mito-Ca 2+ levels using RR and/or histamine in fertilized oocytes and their influence on blastocyst developmental capacity in pigs. Red fluorescence intensity by the mito-Ca 2+ detection dye Rhod-2 was significantly increased (P &lt; 0.05) in zygotes 6 h after IVF compared to mature oocytes. Based on these results, we investigated the changes in mito-Ca 2+ by RR (10 and 20 μM) in presumptive zygotes using Rhod-2 staining and mito-Ca 2+ uptake 1 (MICU1) protein levels as an indicator of mito-Ca 2+ uptake using western blot analysis. As expected, RR-treated zygotes displayed decreased protein levels of MICU1 and Rhod-2 red fluorescence intensity compared to non-treated zygotes 6 h after IVF. Blastocyst development rate of 20 μM RR-treated zygotes was significantly increased 6 h after IVF (P &lt; 0.05) due to improved mitochondrial functions. Conversely, the blastocyst development rate was significantly decreased in histamine (mito-Ca 2+ inhibitor, 100 nM) treated zygotes (P &lt; 0.05). The collective results demonstrate that RR improves blastocyst development in porcine embryos by regulating mito-Ca 2+ and MICU1 expression following IVF.

journal of reproduction and development

Ruthenium red improves blastocyst developmental competence by regulating mitochondrial Ca&lt;sup&gt;2+&lt;/sup&gt; and mitochondrial functions in fertilized porcine oocytes &lt;i&gt;in vitro&lt;/i&gt;