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The negative effect of the PI3K inhibitor 3-methyladenine on planarian regeneration via the autophagy signalling pathway

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Abstract

As an important PI3K (VPS34) inhibitor, 3-methyladenine (3-MA) can block the formation of autophagic vesicles in animals. Most toxicological studies using 3-MA have shown that 3-MA leads to serious disorders via autophagy suppression in mammals. However, no toxicological research on 3-MA has been performed on individuals undergoing regeneration. The freshwater planarian has powerful regenerative capability, and it can regenerate a new brain in 5 days and undergo complete adult individual remodelling in approximately 14 days. Moreover, it is also an excellent model organism for studies on environmental toxicology due to its high chemical sensitivity and extensive distribution. Here, Dugesia japonica planarians were treated with 3-MA, and the results showed that autophagy was inhibited and Djvps34 expression levels were down-regulated. After exposure to 10 mM 3-MA for 18 h, all the controls showed normal phenotypes, while one-half of the planarians treated with 3-MA showed morphological defects. In most cases, an ulcer appeared in the middle of the body, and a normal phenotype was restored 7 days following 3-MA exposure. During regeneration, disproportionate blastemas with tissue regression were observed. Furthermore, 3-MA treatment suppressed stem cell proliferation in intact and regenerating worms. These findings demonstrate that autophagy is indispensable for tissue homeostasis and regeneration in planarians and that 3-MA treatment is detrimental to planarian regeneration via its effect on the autophagy pathway.

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Acknowledgements

We especially thank Dr. Xin Yan from Max-Delbrueck Center in the Helmholtz Association and Dr. Yanli Liu for their critical reviews of this manuscript.

Funding

This work is supported by grants from the National Natural Science Foundation of China (32070427, 31572267, 31471965 and U1604173), the Puyang Field Scientific Observation and Research Station for Yellow River Wetland Ecosystem project of Henan Province, Key Technologies and Program of Henan Province (202102110261 and 202102110383), as well as Major public welfare project of Henan Province (201300311700) and the Science and technology project of Xinxiang City (GG2020012).

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Authors and Affiliations

  1. College of Life Science, Henan Normal University, Xinxiang, China

    Jing Kang, Jinzi Chen, Zimei Dong, Guangwen Chen & Dezeng Liu

  2. College of Life Science, Xingxiang Medical University, Xinxiang, China

    Jing Kang

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  1. Jing Kang
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  2. Jinzi Chen
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Contributions

JK performed TEM, 3-MA treatment,morphological observation and quantitative real-time PCR. JZC performed immunofluorescence. GWC and ZMD conceived of the study, participated in its design and coordination and helped to draft the manuscript. DZL had been involved in revising the manuscript. All authors read and approved the final manuscript.

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Correspondence to Zimei Dong or Guangwen Chen.

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The authors declare no competing interests.

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Informed consent was obtained from all individual participants included in the study.

Research involving human participants and/or animals’ note

This study do not involve endangered or protected species, and the collection of specimens is approved by the Forestry Department of Wild Animal Protection, Henan Province, China.

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Kang, J., Chen, J., Dong, Z. et al. The negative effect of the PI3K inhibitor 3-methyladenine on planarian regeneration via the autophagy signalling pathway. Ecotoxicology 30, 1941–1948 (2021). https://doi.org/10.1007/s10646-021-02439-w

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