PAK1-Dependent Regulation of Microtubule Organization and Spindle Migration Is Essential for the Metaphase I–Metaphase II Transition in Porcine Oocytes
P21-activated kinase 1 (PAK1) is a critical downstream target that mediates the effect of small Rho GTPase on the regulation of cytoskeletal kinetics, cell proliferation, and cell migration. PAK1 has been identified as a crucial regulator of spindle assembly during the first meiotic division; howeve...
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MDPI AG
2024-02-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/14/2/237 |
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| author | Lei Peng Yijing He Weihan Wang Jianjun Dai Qiao Li Shiqiang Ju |
| author_facet | Lei Peng Yijing He Weihan Wang Jianjun Dai Qiao Li Shiqiang Ju |
| author_sort | Lei Peng |
| collection | DOAJ |
| description | P21-activated kinase 1 (PAK1) is a critical downstream target that mediates the effect of small Rho GTPase on the regulation of cytoskeletal kinetics, cell proliferation, and cell migration. PAK1 has been identified as a crucial regulator of spindle assembly during the first meiotic division; however, its roles during the metaphase I (MI) to metaphase II (MII) transition in oocytes remain unclear. In the present study, the potential function of PAK1 in regulating microtubule organization and spindle positioning during the MI–MII transition was addressed in porcine oocytes. The results showed that activated PAK1 was co-localized with α-tubulin, and its expression was increased from the MI to MII stage (<i>p</i> < 0.001). However, inhibiting PAK1 activity with an inhibitor targeting PAK1 activation-3 (IPA-3) at the MI stage decreased the first polar body (PB1) extrusion rate (<i>p</i> < 0.05), with most oocytes arrested at the anaphase-telophase (ATI) stage. IPA-3-treated oocytes displayed a decrease in actin distribution in the plasma membrane (<i>p</i> < 0.001) and an increase in the rate of defects in MII spindle reassembly with abnormal spindle positioning (<i>p</i> < 0.001). Nevertheless, these adverse effects of IPA-3 on oocytes were reversed when the disulfide bond between PAK1 and IPA-3 was reduced by dithiothreitol (DTT). Co-immunoprecipitation revealed that PAK1 could recruit activated Aurora A and transform acidic coiled-coil 3 (TACC3) to regulate spindle assembly and interact with LIM kinase 1 (LIMK1) to facilitate actin filament-mediated spindle migration. Together, PAK1 is essential for microtubule organization and spindle migration during the MI–MII transition in porcine oocytes, which is associated with the activity of p-Aurora A, p-TACC3 and p-LIMK1. |
| first_indexed | 2024-03-07T22:40:25Z |
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| language | English |
| last_indexed | 2024-03-07T22:40:25Z |
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| series | Biomolecules |
| spelling | doaj.art-272cc5294fd3406dae471d8f225fc7392024-02-23T15:09:26ZengMDPI AGBiomolecules2218-273X2024-02-0114223710.3390/biom14020237PAK1-Dependent Regulation of Microtubule Organization and Spindle Migration Is Essential for the Metaphase I–Metaphase II Transition in Porcine OocytesLei Peng0Yijing He1Weihan Wang2Jianjun Dai3Qiao Li4Shiqiang Ju5MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaKey Laboratory of Livestock and Poultry Resources (Pig) Evaluation and Utilization, Ministry of Agriculture and Rural Affairs, Institute of Animal Husbandry and Veterinary Science, Shanghai Academy of Agricultural Sciences, Shanghai 201106, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaP21-activated kinase 1 (PAK1) is a critical downstream target that mediates the effect of small Rho GTPase on the regulation of cytoskeletal kinetics, cell proliferation, and cell migration. PAK1 has been identified as a crucial regulator of spindle assembly during the first meiotic division; however, its roles during the metaphase I (MI) to metaphase II (MII) transition in oocytes remain unclear. In the present study, the potential function of PAK1 in regulating microtubule organization and spindle positioning during the MI–MII transition was addressed in porcine oocytes. The results showed that activated PAK1 was co-localized with α-tubulin, and its expression was increased from the MI to MII stage (<i>p</i> < 0.001). However, inhibiting PAK1 activity with an inhibitor targeting PAK1 activation-3 (IPA-3) at the MI stage decreased the first polar body (PB1) extrusion rate (<i>p</i> < 0.05), with most oocytes arrested at the anaphase-telophase (ATI) stage. IPA-3-treated oocytes displayed a decrease in actin distribution in the plasma membrane (<i>p</i> < 0.001) and an increase in the rate of defects in MII spindle reassembly with abnormal spindle positioning (<i>p</i> < 0.001). Nevertheless, these adverse effects of IPA-3 on oocytes were reversed when the disulfide bond between PAK1 and IPA-3 was reduced by dithiothreitol (DTT). Co-immunoprecipitation revealed that PAK1 could recruit activated Aurora A and transform acidic coiled-coil 3 (TACC3) to regulate spindle assembly and interact with LIM kinase 1 (LIMK1) to facilitate actin filament-mediated spindle migration. Together, PAK1 is essential for microtubule organization and spindle migration during the MI–MII transition in porcine oocytes, which is associated with the activity of p-Aurora A, p-TACC3 and p-LIMK1.https://www.mdpi.com/2218-273X/14/2/237porcine oocytesP21-activated kinase 1metaphase I–metaphase II transitionspindle dynamics |
| spellingShingle | Lei Peng Yijing He Weihan Wang Jianjun Dai Qiao Li Shiqiang Ju PAK1-Dependent Regulation of Microtubule Organization and Spindle Migration Is Essential for the Metaphase I–Metaphase II Transition in Porcine Oocytes Biomolecules porcine oocytes P21-activated kinase 1 metaphase I–metaphase II transition spindle dynamics |
| title | PAK1-Dependent Regulation of Microtubule Organization and Spindle Migration Is Essential for the Metaphase I–Metaphase II Transition in Porcine Oocytes |
| title_full | PAK1-Dependent Regulation of Microtubule Organization and Spindle Migration Is Essential for the Metaphase I–Metaphase II Transition in Porcine Oocytes |
| title_fullStr | PAK1-Dependent Regulation of Microtubule Organization and Spindle Migration Is Essential for the Metaphase I–Metaphase II Transition in Porcine Oocytes |
| title_full_unstemmed | PAK1-Dependent Regulation of Microtubule Organization and Spindle Migration Is Essential for the Metaphase I–Metaphase II Transition in Porcine Oocytes |
| title_short | PAK1-Dependent Regulation of Microtubule Organization and Spindle Migration Is Essential for the Metaphase I–Metaphase II Transition in Porcine Oocytes |
| title_sort | pak1 dependent regulation of microtubule organization and spindle migration is essential for the metaphase i metaphase ii transition in porcine oocytes |
| topic | porcine oocytes P21-activated kinase 1 metaphase I–metaphase II transition spindle dynamics |
| url | https://www.mdpi.com/2218-273X/14/2/237 |
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