Skip to main content
Fig. 6 | Reproductive Biology and Endocrinology

Fig. 6

From: Insufficient HtrA2 causes meiotic defects in aging germinal vesicle oocytes

Fig. 6

HtrA2 overexpression improves oocyte quality in old mice. A Schematic representation of the experimental procedure used to determine whether HtrA2 overexpression could rescue the meiotic defects in oocytes from old mice. B Western blot analysis confirming successful overexpression of exogenous HtrA2 using with an anti-HtrA2 antibody (n = 85 per group). In the overexpression group, approximately 10 pl HtrA2-EGFP mRNA was injected into young GV oocyte. An equal amount of nuclease-free water was injected into young oocytes as a control. (C) Control, Old, Old + NC (RNase-free water), Old+HtrA2FS (encoding frame shifted HtrA2 mRNA) and Old+HtrA2-mRNA (remove EGFP) oocytes were stained with an anti-α-tubulin antibody to visualize the spindle (green) and counterstained with propidium iodide to visualize the chromosomes (red). Arrows point to disorganized spindle (yellow arrows) and misaligned chromosomes (white arrows). Scale bar: 25 μm. D Quantification of Young (n = 60), Old (n = 45), Old+NC (n = 36), Old+HtrA2FS (n = 20), and Old+HtrA2-mRNA (n = 60) oocytes with spindle/chromosome defects. E Chromosome spreading of Young, Old, Old+NC, Old+HtrA2FS and Old+HtrA2-mRNA oocytes. Chromosomes were stained with Hoechst 33342 (purple). Scale bar: 10 μm. F Quantification of aneuploidy in Young (n = 28), Old (n = 33), Old+NC (n = 24), Old+HtrA2FS (n = 13), and Old+HtrA2-mRNA (n = 32) oocytes. Data are expressed as the mean ± SEM of three independent experiments. *, p < 0.05. HtrA2-KD, use siRNA-1

Back to article page