Inhibition of Mitochondrial Fission Alleviates Zearalenone-Induced Mitochondria-Associated Endoplasmic Reticulum Membrane Dysfunction in Piglet Sertoli Cells

Abstract

This study aimed to investigate the effects of zearalenone (ZEA) on piglet Sertoli cell (SC)-mitochondria-associated endoplasmic reticulum (ER) membranes (MAMs) based on mitochondrial fission, and to explore the molecular mechanism of ZEA-induced cell damage. After the SCs were exposed to the ZEA, the cell viability decreased, the Ca2+ levels increased, and the MAM showed structural damage. Moreover, glucose-regulated protein 75 (Grp75) and mitochondrial Rho-GTPase 1 (Miro1) were upregulated at the mRNA and protein levels. However, phosphofurin acidic cluster protein 2 (PACS2), mitofusin2 (Mfn2), voltage-dependent anion channel 1 (VDAC1), and inositol 1,4,5-trisphosphate receptor (IP3R) were downregulated at the mRNA and protein levels. A pretreatment with mitochondrial division inhibitor 1 (Mdivi-1) decreased the ZEA-induced cytotoxicity toward the SCs. In the ZEA + Mdivi-1 group, the cell viability increased, the Ca2+ levels decreased, the MAM damage was repaired, and the expression levels of Grp75 and Miro1 decreased, while those of PACS2, Mfn2, VDAC1, and IP3R increased compared with those in the ZEA-only group. Thus, ZEA causes MAM dysfunction in piglet SCs through mitochondrial fission, and mitochondria can regulate the ER via MAM.

Zearalenone Induces Blood-Testis Barrier Damage through Endoplasmic Reticulum Stress-Mediated Paraptosis of Sertoli Cells in Goats

“…Moreover, both IHC and Western blot analyses showed a decreased expression of BTB junction proteins in ZEA-treated testicular tissue, which was consistent with previous reports in mice [ 22 , 23 ]. In addition, considerable studies have also indicated that ZEA treatment, when in vitro, can hinder the survival of SCs and destroy the structure of the BTB through different signaling pathways to cause testicular toxicity [ 13 , 24 , 45 , 46 ]. In mammalian testes, SCs are the major somatic cells of the seminiferous tubules and play crucial roles in spermatogenesis and the regulation of testicular function [ 47 , 48 ].…”

Combination of Zearalenone and Deoxynivalenol Induces Apoptosis by Mitochondrial Pathway in Piglet Sertoli Cells: Role of Endoplasmic Reticulum Stress

“…Such effects include reductions in mitochondrial membrane potential, along with the liberation of Cyt C and apoptosis-inducing factor (AIF) [ 35 ]. Mfn2 is primarily found in the outer mitochondrial membrane (OMM) and plays a vital role in the establishment and maintenance of MAM structures [ 13 ]. Localizing IFN2 through the ER has been found to induce mitochondrial fission via direct binding to Drp1 and the promotion of Drp1 oligomerization [ 36 ].…”

“…In addition, the tumor suppressor inhibits the anti-apoptotic factor Bcl-2 in response to DNA damage [ 41 ]. There is evidence from previous studies conducted within our research group suggesting that ZEA has the potential to cause damage to piglet SCs through the mitochondrial fission pathway, while the ER can regulate mitochondria through MAMs [ 13 , 14 ]. This study is based on the previous foundation of in-depth research; according to our WB and qRT-PCR results, in the Z + D group, there was a significant increase in the gene and protein expression levels of Bid , P53 , and Caspase -9.…”

“…Cells were treated with varying concentrations of ZEA (0, 15, 30, 45, 60, 80 μM) and DON (0, 0.4, 0.8, 1.2, 1.8, 2.4, 4.8 μM) for 24 h. Prior to exposure to ZEA and DON, cells were pretreated with 4-PBA for 2 h. In subsequent experiments, 1.5 μM 4-PBA and a combination of 30 μM ZEA and 1.2 μM DON were used. The details of the methods of cell culture can be found in Ma et al [ 13 ].…”

“…Our previous research found that ZEA increases the distance of MAM and disrupts its structure and function through the mitochondrial pathway/ERS pathway, ultimately leading to increased intracellular Ca 2+ levels. Pretreatment with a mitochondrial division inhibitor (Mdivi-1)/4-PBA alleviated the abnormal protein expression of each pathway induced by ZEA in MAM [ 13 , 14 ]. However, the specific effects of MAM regulation on apoptosis indicators related to the mitochondrial pathway were not investigated.…”