Aminocarb Exposure Induces Cytotoxicity and Endoplasmic Reticulum Stress-Mediated Apoptosis in Mouse Sustentacular Sertoli Cells: Implications for Male Infertility and Environmental Health

abstract

Simple Summary Exposure to pesticides poses a significant threat to male fertility by compromising crucial cells involved in spermatogenesis. Although aminocarb is a widely used carbamate insecticide, little is known about its impact on male fertility and, consequently, on sustentacular Sertoli cells. In this study, we investigated the effects of increasing concentrations of aminocarb on a mouse Sertoli cell line, TM4. Assessments included analyses of cellular proliferation and viability, membrane integrity, mitochondrial biogenesis and membrane potential and expression and the activity of apoptotic proteins, as well as oxidative stress. Our findings revealed a dose-dependent reduction in the proliferation and viability of TM4 cells following exposure to aminocarb. Moreover, exposure to 5 mu M of aminocarb induced depolarization of mitochondria membrane potential, and a decrease in the ratio of phosphorylated eIF2 alpha to total eIF2 alpha, suggesting heightened endoplasmic reticulum stress via the activation of the eIF2 alpha pathway. An elevation in both caspase-3 protein levels and activity was also observed for the cells under this same aminocarb concentration, thus indicating an apoptotic induction. Altogether, our results demonstrate that aminocarb serves as an apoptotic inducer for mouse sustentacular Sertoli cells in vitro, suggesting its potential to modulate independent pathways of the apoptotic cascade.Abstract Exposure to pesticides, poses a significant threat to male fertility by compromising crucial cells involved in spermatogenesis. Aminocarb, is a widely used carbamate insecticide, although its detrimental effects on the male reproductive system, especially on sustentacular Sertoli cells, pivotal for spermatogenesis, remains poorly understood. In this study, we investigated the effects of escalating concentrations of aminocarb on a mouse Sertoli cell line, TM4. Assessments included cytotoxic analysis, mitochondrial biogenesis and membrane potential, expression of apoptotic proteins, caspase-3 activity, and oxidative stress evaluation. Our findings revealed a dose-dependent reduction in the proliferation and viability of TM4 cells following exposure to increasing concentrations of aminocarb. Notably, exposure to 5 mu M of aminocarb induced depolarization of mitochondria membrane potential, and a significant decrease in the ratio of phosphorylated eIF2 alpha to total eIF2 alpha, suggesting heightened endoplasmic reticulum stress via the activation of the eIF2 alpha pathway. Moreover, the same aminocarb concentration was demonstrated to increase both caspase-3 protein levels and activity, indicating an apoptotic induction. Collectively, our results demonstrate that aminocarb serves as an apoptotic inducer for mouse sustentacular Sertoli cells in vitro, suggesting its potential to modulate independent pathways of the apoptotic cascade. These findings underscore the deleterious impact of aminocarb on spermatogenic performance and male fertility, highlighting the urgent need for further investigation into its mechanisms of action and mitigation strategies to safeguard male fertility.

keywords

SIGNALING PATHWAYS; ACTIVATION; TOXICITY

subject category

Life Sciences & Biomedicine - Other Topics

authors

Moreira, S; Martins, AD; Alves, MG; Pastor, LM; Seco-Rovira, V; Oliveira, PF; Pereira, MD

Groups

G5 - Biomimetic, Biological and Living Materials

Projects

CICECO - Aveiro Institute of Materials (UIDB/50011/2020)

CICECO - Aveiro Institute of Materials (UIDP/50011/2020)

Associated Laboratory CICECO-Aveiro Institute of Materials (LA/P/0006/2020)

acknowledgements

This research was funded by "Fundac & atilde;o para a Ciencia e a Tecnologia"-FCT to CICECO-Aveiro Institute of Materials (UIDB/50011/2020, UIDP/50011/2020) and LA/P/0006/2020, financed by national funds through the FCT/ME (PIDDAC), LAQV-REQUIMTE (UIDB/50006/2020), the post-graduation student S. Moreira (DFA/BD/6438/2020), and junior researcher A.D. Martins (2022.05444.CEECIND). Pedro F. Oliveira (CEECINST/00026/2018) and Marco G. Alves (CDL-CTTRI-267-SGRH/2022) are funded by national funds through FCT-Fundac & atilde;o para a Ciencia e a Tecnologia, I.P., under the Scientific Employment Stimulus-Institutional Call, financed by national funds through the FCT/MCTES/FSE/UE. This work was also supported by FCT-Fundac & atilde;o para a Ciencia e Tecnologia, I.P. by project reference UIDB/04501/2020 and DOI identifier https://doi.org/10.54499/UIDB/04501/2020 and project reference UIDP/04501/2020 and DOI identifier https://doi.org/10.54499/UIDP/04501/2020.