Kinesin KIF15 regulates tubulin acetylation and spindle assembly checkpoint in mouse oocyte meiosis

Zou, Yuan-Jing; Shan, Meng-Meng; Wan, Xiang; Liu, Jing-Cai; Zhang, Kun-Huan; Ju, Jia-Qian; Sun, Shao‐Chen

doi:10.1007/s00018-022-04447-3

Cited by 23 publications

(9 citation statements)

References 43 publications

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“…Following this, protein A and G beads (5 μL) were used to treat the lysates at 4 °C for 3 h. The immunoprecipitate was then resuspended with a 1×SDS loading buffer (20 μL), followed by immunoblotting. Based on the previous report [ 45 ], the IP-PAK1 group was the cell lysate incubated with rabbits polyclonal anti-PAK1 antibody. Additionally, the input group was the cell lysate without anti-PAK1 antibody, and the control group was the cell lysate incubated with rabbit IgG antibody.…”

Section: Methodsmentioning

confidence: 99%

PAK1 Is Involved in the Spindle Assembly during the First Meiotic Division in Porcine Oocytes

Peng

Wang

et al. 2023

IJMS

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P21-activated kinase 1 (PAK1), as a member of the PAK family, has been implicated in various functions during somatic mitosis; however, less is known about its role during oocyte meiosis. Herein, we highlight the indispensable role of PAK1 in regulating spindle assembly and cell cycle progression during the first meiotic division of porcine oocytes. First, we found that the activated PAK1 expressed dynamically, and its subcellular localization was tightly associated with the spindle dynamics during meiosis in porcine oocytes. Specific inhibition of PAK1 activity by inhibitor targeting PAK1 activation-3 (IPA-3) led to impaired extrusion of the first polar body (PB1); with most of the IPA-3-treated oocytes arrested at germinal vesicle breakdown (GVBD) and subjected to failure of bipolar spindle formation. However, the adverse effects caused by IPA-3 on oocytes could be restored by reducing disulfide bonds between PAK1 and IPA-3 with dithiothreitol (DTT) treatment. Furthermore, the co-immunoprecipitation assay revealed that PAK1 interacted directly with Aurora A and transforming acidic coiled coil 3 (TACC3), providing an additional explanation for the similar localization of Aurora A and activated PAK1. Additionally, inhibiting the activity of PAK1 decreased the expression of p-Aurora A and p-TACC3; however, the reduced activity of Aurora A and TACC3 could be restored by DTT. In conclusion, PAK1 plays a crucial role in the proper assembly of the spindle during the first meiotic division of porcine oocytes, and the regulation of PAK1 is associated with its effects on p-Aurora A and p-TACC3 expression.

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Section: Methodsmentioning

confidence: 99%

PAK1 Is Involved in the Spindle Assembly during the First Meiotic Division in Porcine Oocytes

Peng

Wang

et al. 2023

IJMS

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“…Earlier studies indicated that reduced tubulin acetylation affected microtubule stability, in spite of this, high tubulin acetylation did not stabilize microtubules 57 . Altered acetylation level of tubulin leads to defective spindle assembly and chromosome misalignment in mouse oocyte meiosis 58 . Increased tubulin acetylation by KIF17 affect microtubule stability during oocyte meiosis, 59 but reduced tubulin acetylation by RASSF1A also impaired microtubule stability 60 .…”

Section: Discussionmentioning

confidence: 99%

“…57 Altered acetylation level of tubulin leads to defective spindle assembly and chromosome misalignment in mouse oocyte meiosis. 58 Increased tubulin acetylation by KIF17 affect microtubule stability during oocyte meiosis, 59 but reduced tubulin acetylation by RASSF1A also impaired microtubule stability. 60 It remains to be determined how αtubulin hyperacetylation disrupts spindle assembly and chromosome movement during oocyte meiosis, potentially affecting microtubule stability and K-MT interactions.…”

Section: Discussionmentioning

confidence: 99%

KAT8 functions in redox homeostasis and mitochondrial dynamics during mouse oocyte meiosis progression

Wu,

Wang,

Guo

et al. 2024

The FASEB Journal

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As a histone acetyltransferase, lysine acetyltransferase 8 (KAT8) participates in diverse biological processes. However, the effect of KAT8 on oocyte maturation in mice remains unclear. In this study, we found that mouse oocytes overexpressing Kat8‐OE induced maturation failure manifested reduced rates of GVBD and first polar body emission. In addition, immunostaining results revealed that Kat8 overexpressing oocytes showed inappropriate mitochondrial distribution patterns, overproduction of reactive oxygen species (ROS), accumulation of phosphorylated γH2AX, hyperacetylation of α‐tubulin, and severely disrupted spindle/chromosome organization. Moreover, we revealed that Kat8 overexpression induced a decline in SOD1 proteins and KAT8's interaction with SOD1 in mouse ovaries via immunoprecipitation. Western blotting data confirmed that Kat8‐OE induced downregulation of SOD1 expression, which is a key factor for the decline of oocyte quality in advanced maternal age. Also, the injection of Myc‐Sod1 cRNA could partially rescue maternal age‐induced meiotic defects in oocytes. In conclusion, our data demonstrated that high level of KAT8 inhibited SOD1 activity, which in turn induced defects of mitochondrial dynamics, imbalance of redox homeostasis, and spindle/chromosome disorganization during mouse oocyte maturation.

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“…Western blotting protocol was adopted based on previous descriptions [ 40 ]. About 150–200 oocytes were collected for western blotting detection.…”

Section: Methodsmentioning

confidence: 99%

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

Hu,

Liao,

Liu

et al. 2024

Biol Direct

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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.

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Kinesin KIF15 regulates tubulin acetylation and spindle assembly checkpoint in mouse oocyte meiosis

Cited by 23 publications

References 43 publications

PAK1 Is Involved in the Spindle Assembly during the First Meiotic Division in Porcine Oocytes

PAK1 Is Involved in the Spindle Assembly during the First Meiotic Division in Porcine Oocytes

KAT8 functions in redox homeostasis and mitochondrial dynamics during mouse oocyte meiosis progression

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

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