Research Paper Volume 14, Issue 22 pp 9186—9199

Restraint stress of female mice during oocyte development facilitates oocyte postovulatory aging

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Figure 4. Effects of FRSOD on ROS level, mitochondrial membrane potential (MMP), reduced glutathione (GSH)/oxidized glutathione (GSSG) ratio and the expression of antioxidant genes in postovulatory aging oocytes. (A) Shows relative ROS level (average fluorescence intensity value, AFIV) in control (Ctrl) and stressed (Strs) oocytes after aging for 0, 6 or 12 h. Each treatment was repeated 3–7 times with each replicate including 30 oocytes from 2 mice. (B) Contains fluorescence microscopic images showing the AFIV of ROS in Ctrl or Strs oocytes aging for 0 or 12 h. Bar is 160 μm and is applied to all images. (C) Shows MMP (red/green fluorescence intensity) in 12 h-aged oocytes as determined by staining with MMP-specific probe JC-1. (D) Contains fluorescence microscopic images showing JC-1 staining intensity in Ctrl or Strs oocytes aging for 12 h. The same oocytes were observed in Cy3 channel (570 nm, red fluorescence) and in FITC channel (512 nm, green fluorescence), respectively. The JC1-red and JC1-green pictures were merged to compared JC1 red and green ratio. Bar is 15 μm and is applied to all images. (E) Compares the GSH: GSSH ratio in 12 h-aged oocytes between Ctrl and Strs groups. Each treatment was repeated 3 times with each replicate including about 40 oocytes from 2 mice. (F) Shows relative mRNA levels of catalase (Cat), superoxide dismutase 1 (Sod1) and sirtuin 1 (Sirt1) in Ctrl or Strs 12 h-aged oocytes. Each treatment was repeated 3 times with each replicate containing about 30–60 oocytes from 2 mice. *Indicate significant differences (P < 0.05) from values in control mice.