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. 2024 Apr 23;19(1):29.
doi: 10.1186/s13062-024-00471-4.

Loss of AMPK activity induces organelle dysfunction and oxidative stress during oocyte aging

Lin-Lin Hu #  1 , Mei-Hua Liao #  1 , Ya-Xi Liu  2 , Chun-Hua Xing  2 , Lan-Lan Nong  1 , Feng-Lian Yang  3 , Shao-Chen Sun  4   5
Affiliations

Loss of AMPK activity induces organelle dysfunction and oxidative stress during oocyte aging

Lin-Lin Hu et al. Biol Direct. .

Abstract

Background: Oocyte quality is critical for the mammalian reproduction due to its necessity on fertilization and early development. During aging, the declined oocytes showing with organelle dysfunction and oxidative stress lead to infertility. AMP-activated protein kinase (AMPK) is a serine/threonine protein kinase which is important for energy homeostasis for metabolism. Little is known about the potential relationship between AMPK with oocyte aging.

Results: In present study we reported that AMPK was related with low quality of oocytes under post ovulatory aging and the potential mechanism. We showed the altered AMPK level during aging and inhibition of AMPK activity induced mouse oocyte maturation defect. Further analysis indicated that similar with its upstream regulator PKD1, AMPK could reduce ROS level to avoid oxidative stress in oocytes, and this might be due to its regulation on mitochondria function, since loss of AMPK activity induced abnormal distribution, reduced ATP production and mtDNA copy number of mitochondria. Besides, we also found that the ER and Golgi apparatus distribution was aberrant after AMPK inhibition, and enhanced lysosome function was also observed.

Conclusions: Taken together, these data indicated that AMPK is important for the organelle function to reduce oxidative stress during oocyte meiotic maturation.

Keywords: AMPK; Meiosis; Mitochondria; Oocyte; Oxidative stress.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
Expression and localization of AMPK in mouse oocytes. (A) The AMPK protein expression in metaphase I and metaphase II of mouse oocytes by western blotting. 200 oocytes were used for each sample. (B) The localization of AMPK during mouse oocyte maturation. AMPK localized in the cytoplasm of mouse oocytes from GV and MI stage, while there was no AMPK signal in the negative control groups. Green, AMPK; blue, DNA. Bar = 20 μm. (C) The AMPK protein expression in metaphase II of mouse oocytes during aging. 150 oocytes were used for each sample. **P < 0.05. (D) The localization of AMPK during mouse oocyte aging. In the aged oocytes, there was no difference for the AMPK localization, however, the fluorescence signals were much weaker than the fresh oocytes. Green, AMPK; blue, DNA. Bar = 20 μm. (E) The rate of relative intensity of AMPK. **P < 0.05
Fig. 2
Fig. 2
AMPK activity is essential for mouse oocyte maturation quality. (A) The representative images of MII oocyte morphology in the control, aging, aging + COM C groups. The oocytes showed normal polar body in the control group; however, there were fragment oocytes after maturation in the aging oocytes; while more proportion of oocytes in the aging oocytes treated with AMPK inhibitor COM C. Bar = 50 μm. (B) The percentage of fragmentation of oocytes in the control, aging, aging + COM C groups. *p < 0.05. (C) The representative images of oocytes in the control and COM C groups. Bar = 20 μm. (D) The percentage of polar body extrusion in the oocytes of the control and COM C groups. *p < 0.05
Fig. 3
Fig. 3
AMPK suppresses oxidative stress in mouse oocytes. (A) The representative images of ROS level in the oocytes of the control and COM C groups. Green, ROS. Bar = 100 μm. (B) The relative fluorescence intensity of ROS in the control and COM C groups. ***P < 0.001. (C) The representative images of ROS level in the oocytes of the control and PKD1 inhibition groups. Green, ROS. Bar = 100 μm. (D) The relative fluorescence intensity of ROS in the control and PKD1 inhibition groups. *p < 0.05, **P < 0.01
Fig. 4
Fig. 4
AMPK regulates mitochondria functions in mouse oocytes. (A) The representative images of mitochondria distribution in the oocytes after AMPK inhibition. Red, mitochondria; blue, DNA. Bar = 20 μm. (B) The rate of abnormal mitochondria distribution in the oocytes after AMPK inhibition. **p < 0.01. (C) The relative intensity of mitochondria in the oocytes after AMPK inhibition. *p < 0.05. (D) The relative ATP production ratio in the oocytes after AMPK inhibition. **p < 0.01. (E) The relative mtDNA copy number in the oocytes after AMPK inhibition. *p < 0.05
Fig. 5
Fig. 5
AMPK maintains ER and Golgi apparatus in mouse oocytes. (A) The representative images of ER distribution in the oocytes after AMPK inhibition. Blue, ER. Bar = 20 μm. (B) The relative ER fluorescence intensity in the oocytes after AMPK inhibition. ***p < 0.001. (C) The representative images of Golgi apparatus distribution in the oocytes after AMPK inhibition. Green, Golgi apparatus; blue, DNA. Bar = 20 μm. (D) The relative Golgi apparatus fluorescence intensity in the oocytes after AMPK inhibition. ***p < 0.001. (E) The representative images of lysosome distribution in the oocytes after AMPK inhibition. Red, lysosome; blue, DNA. Bar = 20 μm. (F) The relative lysosome fluorescence intensity in the oocytes after AMPK inhibition. **p < 0.01
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