Effects of Insulin on Proliferation, Apoptosis, and Ferroptosis in Primordial Germ Cells via PI3K-AKT-mTOR Signaling Pathway

Liu Ye; Xin Liu; Kai Jin; Yingjie Niu; Qisheng Zuo; Jiuzhou Song; Wei Han; Guohong Chen; Bichun Li

doi:10.3390/genes14101975

Effects of Insulin on Proliferation, Apoptosis, and Ferroptosis in Primordial Germ Cells via PI3K-AKT-mTOR Signaling Pathway

Genes (Basel). 2023 Oct 22;14(10):1975. doi: 10.3390/genes14101975.

Authors

Liu Ye^{1

2

3}, Xin Liu^{1

2

3}, Kai Jin^{1

2

3}, Yingjie Niu^{1

2

3}, Qisheng Zuo^{1

2

3}, Jiuzhou Song⁴, Wei Han⁵, Guohong Chen^{1

2

3}, Bichun Li^{1

2

3

6}

Affiliations

¹ Key Laboratory of Animal Breeding Reproduction and Molecular Design for Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.
² Institutes of Agricultural Science and Technology Development, Yangzhou University, Yangzhou 225009, China.
³ Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou 225009, China.
⁴ Animal & Avian Sciences, University of Maryland, College Park, MA 20742, USA.
⁵ Poultry Institute, Chinese Academy of Agricultural Sciences/Poultry Institute of Jiangsu, Yangzhou 225003, China.
⁶ College of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, China.

Abstract

Primordial germ cells (PGCs) are essential for the genetic modification, resource conservation, and recovery of endangered breeds in chickens and need to remain viable and proliferative in vitro. Therefore, there is an urgent need to elucidate the functions of the influencing factors and their regulatory mechanisms. In this study, PGCs collected from Rugao yellow chicken embryonic eggs at Day 5.5 were cultured in media containing 0, 5, 10, 20, 50, and 100 μg/mL insulin. The results showed that insulin regulates cell proliferation in PGCs in a dose-dependent way, with an optimal dose of 10 μg/mL. Insulin mediates the mRNA expression of cell cycle-, apoptosis-, and ferroptosis-related genes. Insulin at 50 μg/mL and 100 μg/mL slowed down the proliferation with elevated ion content and GSH/oxidized glutathione (GSSG) in PGCs compared to 10 μg/mL. In addition, insulin activates the PI3K/AKT/mTOR pathway dose dependently. Collectively, this study demonstrates that insulin reduces apoptosis and ferroptosis and enhances cell proliferation in a dose-dependent manner via the PI3K-AKT-mTOR signaling pathway in PGCs, providing a new addition to the theory of the regulatory role of the growth and proliferation of PGC in vitro cultures.

Keywords: PGCs; PI3K-AKT-mTOR; apoptosis; ferroptosis; insulin; proliferation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Apoptosis
Cell Proliferation
Chick Embryo
Chickens / metabolism
Ferroptosis*
Germ Cells / metabolism
Insulin / metabolism
Insulin / pharmacology
Phosphatidylinositol 3-Kinases / genetics
Phosphatidylinositol 3-Kinases / metabolism
Proto-Oncogene Proteins c-akt* / genetics
Proto-Oncogene Proteins c-akt* / metabolism
Signal Transduction
TOR Serine-Threonine Kinases / genetics
TOR Serine-Threonine Kinases / metabolism

Substances

Proto-Oncogene Proteins c-akt
Phosphatidylinositol 3-Kinases
Insulin
TOR Serine-Threonine Kinases

Grants and funding

This study was supported by the National Natural Science Foundation of China (32272858, 32172718 and 32202655), National Key R&D Program of China (2021YFD1200301), the China Postdoctoral Science Foundation (2022M722697), the JBGS Project of Seed Industry Revitalization in Jiangsu Province (JBGS (2021) 029) and the Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions for funding the study.