Adenosine Monophosphate-Activated Protein Kinase (AMPK) Phosphorylation Is Required for 20-Hydroxyecdysone Regulates Ecdysis in <i>Apolygus lucorum</i>
The plant mirid bug <i>Apolygus lucorum</i> is an omnivorous pest that can cause considerable economic damage. The steroid hormone 20-hydroxyecdysone (20E) is mainly responsible for molting and metamorphosis. The adenosine monophosphate-activated protein kinase (AMPK) is an intracellular...
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MDPI AG
2023-05-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/10/8587 |
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| author | Yongan Tan Liubin Xiao Jing Zhao Jieyu Zhang Sheraz Ahmad Dejin Xu Guangchun Xu Linquan Ge |
| author_facet | Yongan Tan Liubin Xiao Jing Zhao Jieyu Zhang Sheraz Ahmad Dejin Xu Guangchun Xu Linquan Ge |
| author_sort | Yongan Tan |
| collection | DOAJ |
| description | The plant mirid bug <i>Apolygus lucorum</i> is an omnivorous pest that can cause considerable economic damage. The steroid hormone 20-hydroxyecdysone (20E) is mainly responsible for molting and metamorphosis. The adenosine monophosphate-activated protein kinase (AMPK) is an intracellular energy sensor regulated by 20E, and its activity is regulated allosterically through phosphorylation. It is unknown whether the 20E-regulated insect’s molting and gene expression depends on the AMPK phosphorylation. Herein, we cloned the full-length cDNA of the <i>AlAMPK</i> gene in <i>A. lucorum</i>. <i>AlAMPK</i> mRNA was detected at all developmental stages, whereas the dominant expression was in the midgut and, to a lesser extent, in the epidermis and fat body. Treatment with 20E and AMPK activator 5-aminoimidazole-4-carboxamide-1-β-d-ribofuranoside (AlCAR) or only AlCAR resulted in activation of AlAMPK phosphorylation levels in the fat body, probed with an antibody directed against AMPK phosphorylated at Thr172, enhancing <i>AlAMPK</i> expression, whereas no phosphorylation occurred with compound C. Compared to compound C, 20E and/or AlCAR increased the molting rate, the fifth instar nymphal weight and shortened the development time of <i>A. lucorum</i> in vitro by inducing the expression of <i>EcR-A</i>, <i>EcR-B</i>, <i>USP</i>, and <i>E75-A</i>. Similarly, the knockdown of <i>AlAMPK</i> by RNAi reduced the molting rate of nymphs, the weight of fifth-instar nymphs and blocked the developmental time and the expression of 20E-related genes. Moreover, as observed by TEM, the thickness of the epidermis of the mirid was significantly increased in 20E and/or AlCAR treatments, molting spaces began to form between the cuticle and epidermal cells, and the molting progress of the mirid was significantly improved. These composite data indicated that <i>AlAMPK</i>, as a phosphorylated form in the 20E pathway, plays an important role in hormonal signaling and, in short, regulating insect molting and metamorphosis by switching its phosphorylation status. |
| first_indexed | 2024-03-11T03:40:51Z |
| format | Article |
| id | doaj.art-b65c38dedf1948919e679affa05c5e81 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T03:40:51Z |
| publishDate | 2023-05-01 |
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| spelling | doaj.art-b65c38dedf1948919e679affa05c5e812023-11-18T01:38:21ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-05-012410858710.3390/ijms24108587Adenosine Monophosphate-Activated Protein Kinase (AMPK) Phosphorylation Is Required for 20-Hydroxyecdysone Regulates Ecdysis in <i>Apolygus lucorum</i>Yongan Tan0Liubin Xiao1Jing Zhao2Jieyu Zhang3Sheraz Ahmad4Dejin Xu5Guangchun Xu6Linquan Ge7Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, ChinaInstitute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, ChinaInstitute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, ChinaInstitute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, ChinaCollege of Plant Protection, Yangzhou University, Yangzhou 225009, ChinaInstitute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, ChinaInstitute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, ChinaCollege of Plant Protection, Yangzhou University, Yangzhou 225009, ChinaThe plant mirid bug <i>Apolygus lucorum</i> is an omnivorous pest that can cause considerable economic damage. The steroid hormone 20-hydroxyecdysone (20E) is mainly responsible for molting and metamorphosis. The adenosine monophosphate-activated protein kinase (AMPK) is an intracellular energy sensor regulated by 20E, and its activity is regulated allosterically through phosphorylation. It is unknown whether the 20E-regulated insect’s molting and gene expression depends on the AMPK phosphorylation. Herein, we cloned the full-length cDNA of the <i>AlAMPK</i> gene in <i>A. lucorum</i>. <i>AlAMPK</i> mRNA was detected at all developmental stages, whereas the dominant expression was in the midgut and, to a lesser extent, in the epidermis and fat body. Treatment with 20E and AMPK activator 5-aminoimidazole-4-carboxamide-1-β-d-ribofuranoside (AlCAR) or only AlCAR resulted in activation of AlAMPK phosphorylation levels in the fat body, probed with an antibody directed against AMPK phosphorylated at Thr172, enhancing <i>AlAMPK</i> expression, whereas no phosphorylation occurred with compound C. Compared to compound C, 20E and/or AlCAR increased the molting rate, the fifth instar nymphal weight and shortened the development time of <i>A. lucorum</i> in vitro by inducing the expression of <i>EcR-A</i>, <i>EcR-B</i>, <i>USP</i>, and <i>E75-A</i>. Similarly, the knockdown of <i>AlAMPK</i> by RNAi reduced the molting rate of nymphs, the weight of fifth-instar nymphs and blocked the developmental time and the expression of 20E-related genes. Moreover, as observed by TEM, the thickness of the epidermis of the mirid was significantly increased in 20E and/or AlCAR treatments, molting spaces began to form between the cuticle and epidermal cells, and the molting progress of the mirid was significantly improved. These composite data indicated that <i>AlAMPK</i>, as a phosphorylated form in the 20E pathway, plays an important role in hormonal signaling and, in short, regulating insect molting and metamorphosis by switching its phosphorylation status.https://www.mdpi.com/1422-0067/24/10/8587<i>Apolygus lucorum</i>AMPK20Ephosphorylationecdysis |
| spellingShingle | Yongan Tan Liubin Xiao Jing Zhao Jieyu Zhang Sheraz Ahmad Dejin Xu Guangchun Xu Linquan Ge Adenosine Monophosphate-Activated Protein Kinase (AMPK) Phosphorylation Is Required for 20-Hydroxyecdysone Regulates Ecdysis in <i>Apolygus lucorum</i> International Journal of Molecular Sciences <i>Apolygus lucorum</i> AMPK 20E phosphorylation ecdysis |
| title | Adenosine Monophosphate-Activated Protein Kinase (AMPK) Phosphorylation Is Required for 20-Hydroxyecdysone Regulates Ecdysis in <i>Apolygus lucorum</i> |
| title_full | Adenosine Monophosphate-Activated Protein Kinase (AMPK) Phosphorylation Is Required for 20-Hydroxyecdysone Regulates Ecdysis in <i>Apolygus lucorum</i> |
| title_fullStr | Adenosine Monophosphate-Activated Protein Kinase (AMPK) Phosphorylation Is Required for 20-Hydroxyecdysone Regulates Ecdysis in <i>Apolygus lucorum</i> |
| title_full_unstemmed | Adenosine Monophosphate-Activated Protein Kinase (AMPK) Phosphorylation Is Required for 20-Hydroxyecdysone Regulates Ecdysis in <i>Apolygus lucorum</i> |
| title_short | Adenosine Monophosphate-Activated Protein Kinase (AMPK) Phosphorylation Is Required for 20-Hydroxyecdysone Regulates Ecdysis in <i>Apolygus lucorum</i> |
| title_sort | adenosine monophosphate activated protein kinase ampk phosphorylation is required for 20 hydroxyecdysone regulates ecdysis in i apolygus lucorum i |
| topic | <i>Apolygus lucorum</i> AMPK 20E phosphorylation ecdysis |
| url | https://www.mdpi.com/1422-0067/24/10/8587 |
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