Ethanol-Induced Mitochondrial Damage in Sertoli Cells is Associated with Parkin Overexpression and Activation of Mitophagy
Abstract
:1. Introduction
2. Materials and Methods
2.1. Study Approval
2.2. Antibodies and Kits
2.3. Animals and Experimental Procedure
2.4. IHC for LC3, Parkin, PINK1, TFEB, and iNOS
2.5. IF Single and Double Labeling of Mitophagy Proteins and Mitochondrial and Lysosomal Markers
2.6. Line Profile Plots for Co-Localization Analysis of Parkin and Pan-Cathepsin
2.7. Terminal Deoxynucleotidyl Transferase dUTP-Mediated Nick-End Labeling (TUNEL) Assay
2.8. TEM and Quantitative Analysis of Mitophagic Vacuoles (MVs)
2.9. Immunogold Labeling for LC3, Parkin, and TFEB and Double Immunogold Labeling of Parkin and PINK1
2.10. Western Blot Analysis for LC3, Cytochrome c, Parkin, TFEB, COX IV, and iNOS
2.11. Statistical Analysis
3. Results
3.1. Enhanced Mitochondrial Damage and Mitophagic Vacuole (MV) Formation in ETR SCs with Predominant Localization in Perinuclear Areas
3.2. Association of Ethanol-Induced Mitophagosomes in SCs with Increased LC3-II Expression and Mitochondrial Proteins Reduction
3.3. Ethanol Increased the Expression and Mitochondrial Translocation of Parkin and PINK1 in SCs and Fusion with Lysosomal Compartment
3.4. Enhanced Mitophagy in SCs of ETRs is Associated with TFEB Nuclear Translocation and Induction of iNOS
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
- Kroemer, G.; Marino, G.; Levine, B. Autophagy and the integrated stress response. Mol. Cell 2010, 40, 280–293. [Google Scholar] [CrossRef] [PubMed]
- Eid, N.; Ito, Y.; Otsuki, Y. The autophagic response to alcohol toxicity: The missing layer. J. Hepatol. 2013, 59, 398. [Google Scholar] [CrossRef] [Green Version]
- Sica, V.; Galluzzi, L.; Bravo-San Pedro, J.M.; Izzo, V.; Maiuri, M.C.; Kroemer, G. Organelle-specific initiation of autophagy. Mol. Cell 2015, 59, 522–539. [Google Scholar] [CrossRef]
- Lemasters, J.J. Variants of mitochondrial autophagy: Types 1 and2 mitophagy and micromitophagy (type 3). Redox Biol. 2014, 2, 749–754. [Google Scholar] [CrossRef]
- Ding, W.X.; Li, M.; Chen, X.; Ni, H.M.; Lin, C.W.; Gao, W.; Lu, B.; Stolz, D.B.; Clemens, D.L.; Yin, X.M. Autophagy reduces acute ethanol-induced hepatotoxicity and steatosis in mice. Gastroenterology 2010, 139, 1740–1752. [Google Scholar] [CrossRef] [PubMed]
- Eid, N.; Ito, Y.; Horibe, A.; Otsuki, Y. Ethanol-induced mitophagy in liver is associated with activation of the PINK1-Parkin pathway triggered by oxidative DNA damage. Histol. Histopathol. 2016, 31, 1143–1159. [Google Scholar]
- Eid, N.; Ito, Y.; Otsuki, Y. Triggering of Parkin mitochondrial translocation in mitophagy: Implications for liver diseases. Front. Pharmacol. 2016, 7, 100. [Google Scholar] [CrossRef] [PubMed]
- Eid, N.; Ito, Y.; Horibe, A.; Hamaoka, H.; Kondo, Y. A method for in vivo induction and ultrastructural detection of mitophagy in Sertoli cells. Methods Mol. Biol. 2018, 1748, 103–112. [Google Scholar] [PubMed]
- Eid, N.; Kondo, Y. Ethanol-induced mitophagy in rat Sertoli cells: Implications for male fertility. Andrologia 2018, 50, e12820. [Google Scholar] [CrossRef]
- Vives-Bauza, C.; Zhou, C.; Huang, Y.; Cui, M.; de Vries, R.L.; Kim, J.; May, J.; Tocilescu, M.A.; Liu, W.; Ko, H.S.; et al. PINK1-dependent recruitment of Parkin to mitochondria in mitophagy. Proc. Nat. Acad. Sci. 2010, 107, 378–383. [Google Scholar] [CrossRef]
- Narendra, D.; Tanaka, A.; Suen, D.F.; Youle, R.J. Parkin is recruited selectively to impaired mitochondria and promotes their autophagy. J. Cell Biol. 2008, 183, 795–803. [Google Scholar] [CrossRef] [Green Version]
- Williams, J.A.; Ni, H.M.; Ding, Y.; Ding, W.X. Parkin regulates mitophagy and mitochondrial function to protect against alcohol-induced liver injury and steatosis in mice. Am. J. Physiol. Gastrointest. Liver Physiol. 2015, 309, 324–340. [Google Scholar] [CrossRef]
- Williams, J.A.; Ding, W.X. A mechanistic review of mitophagy and Its role in protection against alcoholic liver disease. Biomolecules 2015, 5, 2619–2642. [Google Scholar] [CrossRef]
- Hwang, C.J.; Kim, Y.E.; Son, D.J.; Park, M.H.; Choi, D.Y.; Park, P.H.; Hellström, M.; Han, S.B.; Oh, K.W.; Park, E.K.; et al. Parkin deficiency exacerbate ethanol-induced dopaminergic neurodegeneration by P38 pathway dependent inhibition of autophagy and mitochondrial function. Redox Biol. 2017, 11, 456–468. [Google Scholar] [CrossRef] [PubMed]
- Dai, Y.; Hu, X.; Sun, X. Overexpression of parkin protects retinal ganglion cells in experimental glaucoma. Cell Death Dis. 2018, 9, 88. [Google Scholar] [CrossRef] [Green Version]
- Horibe, A.; Eid, N.; Ito, Y.; Hamaoka, H.; Tanaka, Y.; Kondo, Y. Upregulated autophagy in Sertoli cells of ethanol-treated rats is associated with induction of inducible nitric oxide synthase (iNOS), androgen receptor suppression and germ cell apoptosis. Int. J. Mol. Sci. 2017, 18, 1061. [Google Scholar] [CrossRef]
- Thomes, P.G.; Trambly, C.S.; Fox, H.S.; Tuma, D.J.; Donohue, T.M., Jr. Acute and chronic ethanol administration differentially modulate hepatic autophagy and transcription factor EB. Alcohol. Clin. Exp. Res. 2015, 39, 2354–2363. [Google Scholar] [CrossRef] [PubMed]
- Tan, S.; Wong, E. Mitophagy transcriptome: Mechanistic insights into polyphenol-mediated mitophagy. Oxid. Med. Cell Longev. 2017, 2017, 9028435. [Google Scholar] [CrossRef] [PubMed]
- D’Souza El-Guindy, N.B.; Kovacs, E.J.; de Witte, P.; Spies, C.; Littleton, J.M.; de Villiers, W.J.; Lott, A.J.; Plackett, T.P.; Lanzke, N.; Meadows, G.G. Laboratory models available to study alcohol-induced organ damage and immune variations: Choosing the appropriate model. Alcohol. Clin. Exp. Res. 2010, 3, 1489–1511. [Google Scholar]
- Nogales, F.; Rua, R.M.; Ojeda, M.L.; Murillo, M.L.; Carreras, O. Oral or intraperitoneal binge drinking and oxidative balance in adolescent rats. Chem. Res. Toxicol. 2014, 27, 1926–1933. [Google Scholar] [CrossRef] [PubMed]
- Narabayashi, K.; Ito, Y.; Eid, N.; Maemura, K.; Inoue, T.; Takeuchi, T.; Otsuki, Y.; Higuchi, K. Indomethacin suppresses LAMP-2 expression and induces lipophagy and lipoapoptosis in rat enterocytes via the ER stress pathway. J. Gastroenterol. 2015, 50, 541–554. [Google Scholar] [CrossRef] [PubMed]
- Horibe, A.; Eid, N.; Ito, Y.; Otsuki, Y.; Kondo, Y. Ethanol-induced autophagy in Sertoli cells is specifically marked at androgen-dependent stages of the spermatogenic cycle: Potential mechanisms and implications. Int. J. Mol. Sci. 2019, 20. [Google Scholar] [CrossRef]
- Amadoro, G.; Corsetti, V.; Florenzano, F.; Atlante, A.; Bobba, A.; Nicolin, V.; Nori, S.L.; Calissano, P. Morphological and bioenergetic demands underlying the mitophagy in post-mitotic neurons: The pinkparkin pathway. Front. Aging Neurosci. 2014, 6, 18. [Google Scholar] [CrossRef] [PubMed]
- Amadoro, G.; Corsetti, V.; Florenzano, F.; Atlante, A.; Ciotti, M.T.; Mongiardi, M.P.; Bussani, R.; Nicolin, V.; Nori, S.L.; Campanella, M.; et al. AD-linked, toxic NH2 human tau affects the quality control of mitochondria in neurons. Neurobiol. Dis. 2014, 62, 489–507. [Google Scholar] [CrossRef]
- Teckman, J.H.; An, J.K.; Blomenkamp, K.; Schmidt, B.; Perlmutter, D. Mitochondrial autophagy and injury in the liver in alpha 1-antitrypsin deficiency. Am. J. Physiol. Gastrointest Liver Physiol. 2004, 286, 851–862. [Google Scholar] [CrossRef]
- Eid, N.; Ito, Y.; Maemura, K.; Otsuki, Y. Elevated autophagic sequestration of mitochondria and lipid droplets in steatotic hepatocytes of chronic ethanol treated rats: An immunohistochemical and electron microscopic study. J. Mol. Histol. 2013, 44, 311–326. [Google Scholar] [CrossRef] [PubMed]
- Lobo, M.V.; Alonso, F.J.; Arenas, M.I.; Caso, E.; Fraile, B.; del Río, R.M. Ultrastructural staining with sodium metaperiodate and sodium borohydride. J. Histochem. Cytochem. 2002, 50, 11–19. [Google Scholar] [CrossRef]
- Bergersen, L.H.; Storm-Mathisen, J.; Gundersen, V. Immunogold quantification of amino acids and proteins in complex subcellular compartments. Nat. Protoc. 2008, 3, 144–152. [Google Scholar] [CrossRef]
- Guo, X.; Sun, X.; Hu, D.; Wang, Y.J.; Fujioka, H.; Vyas, R.; Chakrapani, S.; Joshi, A.U.; Luo, Y.; Mochly-Rosen, D.; et al. VCP recruitment to mitochondria causes mitophagy impairment and neurodegeneration in models of Huntington’s disease. Nat. Commun. 2016, 26, 12646. [Google Scholar] [CrossRef] [PubMed]
- Lim, J.; Kim, H.W.; Youdim, M.B.; Rhyu, I.J.; Choe, K.M.; Oh, Y.J. Binding preference of p62 towardsLC3-ll during dopaminergic neurotoxin-induced impairment of autophagic flux. Autophagy 2011, 7, 51–60. [Google Scholar] [CrossRef]
- Saita, S.; Shirane, M.; Nakayama, K.I. Selective escape of proteins from the mitochondria during mitophagy. Nat. Commun. 2013, 4, 1410. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Flores-Toro, J.A.; Go, K.L.; Leeuwenburgh, C.; Kim, J.S. Autophagy in the liver: Cell’s cannibalism and beyond. Arch Pharm. Res. 2016, 39, 1050–1061. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Oliveira, P.F.; Martins, A.D.; Moreira, A.C.; Cheng, C.Y.; Alves, M.G. The Warburg effect revisited-lesson from the Sertoli cell. Med. Res. Rev. 2015, 35, 126–151. [Google Scholar] [CrossRef] [PubMed]
- Rato, L.; Meneses, M.J.; Silva, B.M.; Sousa, M.; Alves, M.G.; Oliveira, P.F. New insights on hormones and factors that modulate Sertoli cell metabolism. Histol. Histopathol. 2016, 31, 499–513. [Google Scholar]
- Zhang, J. Teaching the basics of autophagy and mitophagy to redox biologists--mechanisms and experimental approaches. Redox Biol. 2015, 4, 242–459. [Google Scholar] [CrossRef]
- Xiao, B.; Goh, J.Y.; Xiao, L.; Xian, H.; Lim, K.L.; Liou, Y.C. Reactive oxygen species trigger Parkin/PINK1 pathway-dependent mitophagy by inducing mitochondrial recruitment of Parkin. J. Biol. Chem. 2017, 292, 16697–16708. [Google Scholar] [CrossRef] [PubMed]
- Kim, S.J.; Syed, G.H.; Siddiqui, A.; Hepatitis, C. Virus Induces the Mitochondrial Translocation of Parkin and Subsequent Mitophagy. PLoS Pathog. 2013, 9, e1003285. [Google Scholar] [CrossRef] [PubMed]
- Yamano, K.; Matsuda, N.; Tanaka, K. The ubiquitin signal and autophagy: An orchestrated dance leading to mitochondrial degradation. EMBO Rep. 2016, 3, 300–316. [Google Scholar] [CrossRef]
- Palikaras, K.; Lionaki, E.; Tavernarakis, N. Mechanisms of mitophagy in cellular homeostasis, physiology and pathology. Nat. Cell Biol. 2018, 20, 1013–1022. [Google Scholar] [CrossRef]
- Raimundo, N.; Fernández-Mosquera, L.; Yambire, K.F.; Diogo, C.V. Mechanisms of communication between mitochondria and lysosomes. Int. J. Biochem. Cell Biol. 2016, 79, 345–349. [Google Scholar] [CrossRef]
- Richburg, J.H.; Murphy, C.; Myers, J.L. The Sertoli cell as a target for toxicants. In Comprehensive Toxicology, 3rd ed.; McQueen, C.A., Ed.; Elsevier: Amsterdam, The Netherlands, 2014; pp. 64–82. [Google Scholar]
- Mohanty, A.; Tiwari-Pandey, R.; Pandey, N.R. Mitochondria: The indispensable players in innate immunity and guardians of the inflammatory response. J. Cell Commun. Signal. 2019. [Google Scholar] [CrossRef]
- Sanchez-Alvarez, R.; Martinez-Outschoorn, U.E.; Lin, Z.; Lamb, R.; Hulit, J.; Howell, A.; Sotgia, F.; Rubin, E.; Lisanti, M.P. Ethanol exposure induces the cancer-associated fibroblast phenotype and lethal tumor metabolism: Implications for breast cancer prevention. Cell Cycle 2013, 12, 289–301. [Google Scholar] [CrossRef] [PubMed]
- Khandelwal, P.J.; Herman, A.M.; Hoe, H.S.; Rebeck, G.W.; Moussa, C.E. Parkin mediates beclin-dependent autophagic clearance of defective mitochondria and ubiquitinated Abeta in AD models. Hum. Mol. Genet. 2011, 20, 2091–2102. [Google Scholar] [CrossRef] [PubMed]
- Fang, E.F.; Hou, Y.; Palikaras, K.; Adriaanse, B.A.; Kerr, J.S.; Yang, B.; Lautrup, S.; Hasan-Olive, M.M.; Caponio, D.; Dan, X.; et al. Mitophagy inhibits amyloid-β and tau pathology and reverses cognitive deficits in models of Alzheimer’s disease. Nat. Neurosci. 2019. [Epub ahead of print]. [Google Scholar] [CrossRef] [PubMed]
- Zhang, S.X.; Zhuang, L.L.; Liu, J.; Jing, Y.Y.; Sun, J.; Gong, L.; Liu, X.Y. The role of Parkin protein in cardiac function and ventricular remodeling in myocardial infarction rats. Eur. Rev. Med. Pharmacol. Sci. 2018, 22, 5004–5013. [Google Scholar] [PubMed]
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Eid, N.; Ito, Y.; Horibe, A.; Otsuki, Y.; Kondo, Y. Ethanol-Induced Mitochondrial Damage in Sertoli Cells is Associated with Parkin Overexpression and Activation of Mitophagy. Cells 2019, 8, 283. https://doi.org/10.3390/cells8030283
Eid N, Ito Y, Horibe A, Otsuki Y, Kondo Y. Ethanol-Induced Mitochondrial Damage in Sertoli Cells is Associated with Parkin Overexpression and Activation of Mitophagy. Cells. 2019; 8(3):283. https://doi.org/10.3390/cells8030283
Chicago/Turabian StyleEid, Nabil, Yuko Ito, Akio Horibe, Yoshinori Otsuki, and Yoichi Kondo. 2019. "Ethanol-Induced Mitochondrial Damage in Sertoli Cells is Associated with Parkin Overexpression and Activation of Mitophagy" Cells 8, no. 3: 283. https://doi.org/10.3390/cells8030283