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A new insight of cadmium-induced cellular evidence of autophagic-associated spermiophagy during spermatogenesis

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Abstract

Autophagy plays a dynamic role in spermatozoa development during spermatogenesis. However, the disruption of autophagic flux induces cell death under metal toxicity and severe oxidative stress. Therefore, we hypothesized that cadmium-induced autophagy might be involved in this mechanism. To verify this hypothesis, we studied cadmium-induced cellular evidence of autophagic-associated spermiophagy within the testis. In the present study, treatment with cadmium caused nuclear depressive disorders and vacuolated mitochondrial damage of Sertoli cells. In addition, spermiophagy through the cellular evidence of spermatozoa phagocytosis, the high lysosomal activity (lysosome engulfment and phagolysosome), and autophagy activity (autolysosome and autophagosome) were observed in the Sertoli cells. The immunohistochemistry of lysosomal membrane protein (LAMP2) to target the phagocytosis of spermatozoa revealed that the immunoreactivity of LAMP2 was overstimulated in the luminal compartment of testis’s seminiferous tubules. In addition, the immunohistochemistry and immunofluorescence of autophagy-related protein and microtubule-associated light chain (LC3) results showed the strong immunoreactivity and immunosignaling of LC3 in the Sertoli cells of the testis. Moreover, cadmium caused the overactivation of the expression level of autophagy-related proteins, autophagy-related gene (ATG7), (ATG5), beclin1, LC3, sequestosome 1 (P62), and LAMP2 which were confirmed by western blotting. In summary, this study demonstrated that hazards related to cadmium-induced autophagic-associated spermiophagy with the disruption of autophagic flux, providing new insights into the toxicity of cadmium in mammals and representing a high risk to male fertility.

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The data sets collated or analyzed during the current study are accessible upon adequate request from the corresponding author.

References

  • Ahmed N, Yang P, Chen H, Ujjan IA, Haseeb A, Wang L, Soomro F, Faraz S, Sahito B, Ali W (2018) Characterization of inter-Sertoli cell tight and gap junctions in the testis of turtle: protect the developing germ cells from an immune response. Microb Pathog 123:60–67

    CAS  Google Scholar 

  • Akbarsha MA, Kadalmani B, Tamilarasan V (2007) Efferent ductules of the fan-throated lizard Sitana ponticeriana Cuvier: light and transmission electron microscopy study. Acta Zool 88:265–274

    Google Scholar 

  • Ali W, Bian Y, Ali H, Sun J, Zhu J, Ma Y, Liu Z, Zou H (2023a) Cadmium-induced impairment of spermatozoa development by reducing exosomal-MVBs secretion: a novel pathway. Aging (Albany NY) 15:4096

    Google Scholar 

  • Ali W, Bian Y, Zhang H, Qazi IH, Zou H, Zhu J, Liu Z (2023b) Effect of cadmium exposure during and after pregnancy of female. Environ Pollut Bioavailab 35:2181124

    Google Scholar 

  • Ali W, Ma Y, Zhu J, Zou H, Liu Z (2022) Mechanisms of cadmium-induced testicular injury: a risk to male fertility. Cells 11:3601

    CAS  Google Scholar 

  • Aparicio IM, Rojo-Domínguez P, Castillejo-Rufo A, Peña FJ, Tapia JA (2023) The autophagy marker LC3 is processed during the sperm capacitation and the acrosome reaction and translocates to the acrosome where it colocalizes with the acrosomal membranes in horse spermatozoa. Int J Mol Sci 24:937

    CAS  Google Scholar 

  • Barratt CL, Björndahl L, De Jonge CJ, Lamb DJ, Osorio Martini F, McLachlan R, Oates RD, van der Poel S, St John B, Sigman M (2017) The diagnosis of male infertility: an analysis of the evidence to support the development of global WHO guidance—challenges and future research opportunities. Hum Reprod Update 23:660–680

    Google Scholar 

  • Chen H, Huang Y, Yang P, Shi Y, Ahmed N, Liu T, Bai X, Haseeb A, Chen Q (2020) Autophagy enhances lipid droplet development during spermiogenesis in Chinese soft-shelled turtle, Pelodiscus sinensis. Theriogenology 147:154–165

    CAS  Google Scholar 

  • Chiba A, Nakamura M, Morimoto G (2011) Spermiophagy in the male reproductive tract of some passerine birds. Zool Sci 28:689–693

    Google Scholar 

  • Duan Y, Zhao Y, Wang T, Sun J, Ali W, Ma Y, Yuan Y, Gu J, Bian J, Liu Z (2023) Taurine alleviates cadmium-induced hepatotoxicity by regulating autophagy flux. Int J Mol Sci 24:1205

    CAS  Google Scholar 

  • Elblehi SS, El Euony OI, El-Nahas AF (2019) Partial ameliorative effect of Moringa leaf ethanolic extract on the reproductive toxicity and the expression of steroidogenic genes induced by subchronic cadmium in male rats. Environ Sci Pollut Res 26:23306–23318

    CAS  Google Scholar 

  • Fouad AA, Albuali WH, Jresat I (2013) Simvastatin treatment ameliorates injury of rat testes induced by cadmium toxicity. Biol Trace Elem Res 153:269–278

    CAS  Google Scholar 

  • Grijmans BJ, van der Kooij SB, Varela M, Meijer AH (2022) LAPped in proof: LC3-associated phagocytosis and the arms race against bacterial pathogens. Front Cell Infect Microbiol 11:809121

    Google Scholar 

  • Haseeb A, Bai X, Tarique I, Chen H, Yang P, Gandahi N, Iqbal A, Huang Y, Chen Q (2019) Characterization of in vivo autophagy during avian spermatogenesis. Poult Sci 98:5089–5099

    CAS  Google Scholar 

  • Hassine MBH, Venditti M, Rhouma MB, Minucci S, Messaoudi I (2023) Combined effect of polystyrene microplastics and cadmium on rat blood-testis barrier integrity and sperm quality. Environ Sci Pollut Res 30:56700–56712

    CAS  Google Scholar 

  • Heckmann BL, Green DR (2019) LC3-associated phagocytosis at a glance. J Cell Sci 132:jcs222984

    CAS  Google Scholar 

  • Herpin A, Englberger E, Zehner M, Wacker R, Gessler M, Schard M (2015) Defective autophagy through epg5 mutation results in failure to reduce germ plasm and mitochondria. FASEB J 29:4145–4161

    CAS  Google Scholar 

  • Huang Q, Liu Y, Zhang S, Yap YT, Li W, Zhang D, Gardner A, Zhang L, Song S, Hess RA (2021) Autophagy core protein ATG5 is required for elongating spermatid development, sperm individualization and normal fertility in male mice. Autophagy 17:1753–1767

    CAS  Google Scholar 

  • Huang Y, Yang P, Chen H, Bai X, Wang X, Vistro WA, Haseeb A, Shi Y, Chen Q (2018) A “Lamellar structure” contributes to autophagosome biogenesis and mitophagy in zebrafish hepatocytes. Fish Shellfish Immunol 81:83–91

    CAS  Google Scholar 

  • Ji Y-L, Wang H, Zhao X-F, Wang Q, Zhang C, Zhang Y, Zhao M, Chen Y-H, Meng X-H, Xu D-X (2011) Crosstalk between endoplasmic reticulum stress and mitochondrial pathway mediates cadmium-induced germ cell apoptosis in testes. Toxicol Sci 124:446–459

    CAS  Google Scholar 

  • Klionsky DJ, Abdel-Aziz AK, Abdelfatah S, Abdellatif M, Abdoli A, Abel S, Abeliovich H, Abildgaard MH, Abudu YP, Acevedo-Arozena A (2021) Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy 17:1–382

    Google Scholar 

  • Lei Y, Zhang X, Xu Q, Liu S, Li C, Jiang H, Lin H, Kong E, Liu J, Qi S (2021) Autophagic elimination of ribosomes during spermiogenesis provides energy for flagellar motility. Dev Cell 56(2313-2328):e7

    Google Scholar 

  • Levine H, Jørgensen N, Martino-Andrade A, Mendiola J, Weksler-Derri D, Jolles M, Pinotti R, Swan SH (2023) Temporal trends in sperm count: a systematic review and meta-regression analysis of samples collected globally in the 20th and 21st centuries. Hum Reprod Update 29:157–176

    Google Scholar 

  • Lu Y, Agarwal A (2022) Myo-inositol oxygenase in cadmium-induced kidney injury. American Physiological Society Rockville, MD, pp F470–F472

    Google Scholar 

  • Lv C, Wang X, Guo Y, Yuan S (2020) Role of selective autophagy in spermatogenesis and male fertility. Cells 9:2523

    CAS  Google Scholar 

  • Ma Y, Su Q, Yue C, Zou H, Zhu J, Zhao H, Song R, Liu Z (2022) The effect of oxidative stress-induced autophagy by cadmium exposure in kidney, liver, and bone damage, and neurotoxicity. Int J Mol Sci 23:13491

    CAS  Google Scholar 

  • Niture S, Lin M, Qi Q, Moore JT, Levine KE, Fernando RA, Kumar D (2021) Role of autophagy in cadmium-induced hepatotoxicity and liver diseases. J Toxicol 2021

  • Nogueira KOPC, Araújo VA, Sartori SSR, Neves CA (2011) Phagocytosis of spermatozoa by epithelial cells in the vagina of the lizard Hemidactylus mabouia (Reptilia, Squamata). Micron 42:377–380

    Google Scholar 

  • Ozturk N, Steger K, Schagdarsurengin U (2017) The impact of autophagy in spermiogenesis. Asian J Androl 19:617

    CAS  Google Scholar 

  • Pierron F, Daffe G, Daramy F, Heroin D, Barré A, Bouchez O, Clérendeau C, Romero-Ramirez A, Nikolski M (2023) Transgenerational endocrine disruptor effects of cadmium in zebrafish and contribution of standing epigenetic variation to adaptation. J Hazard Mater 455:131579

    CAS  Google Scholar 

  • Porawski M, Wassermann G, Achaval M (2004) Localization of acid phosphatase activity in the testis of two teleostean species (Oreochromis niloticus and Odonthestes perugiae). Braz J Biol 64:853–858

    CAS  Google Scholar 

  • Ran D, Zhou D, Liu G, Ma Y, Ali W, Yu R, Wang Q, Zhao H, Zhu J, Zou H (2023) Reactive oxygen species control osteoblast apoptosis through SIRT1/PGC-1α/P53Lys382 signaling, mediating the onset of Cd-induced osteoporosis. J Agric Food Chem 71:5991–6002

    CAS  Google Scholar 

  • Roan NR, Sandi-Monroy N, Kohgadai N, Usmani SM, Hamil KG, Neidleman J, Montano M, Ständker L, Röcker A, Cavrois M (2017) Semen amyloids participate in spermatozoa selection and clearance. Elife 6:e24888

    Google Scholar 

  • Sanchez-Vera V, Kenchappa CS, Landberg K, Bressendorff S, Schwarzbach S, Martin T, Mundy J, Petersen M, Thelander M, Sundberg E (2017) Autophagy is required for gamete differentiation in the moss Physcomitrella patens. Autophagy 13:1939–1951

    CAS  Google Scholar 

  • Shang Y, Wang H, Jia P, Zhao H, Liu C, Liu W, Song Z, Xu Z, Yang L, Wang Y (2016) Autophagy regulates spermatid differentiation via degradation of PDLIM1. Autophagy 12:1575–1592

    CAS  Google Scholar 

  • Sharma A, Minhas S, Dhillo WS, Jayasena CN (2021) Male infertility due to testicular disorders. J Clin Endocrinol Metabol 106:e442–e459

    Google Scholar 

  • Sharma A, Mollier J, Brocklesby RW, Caves C, Jayasena CN, Minhas S (2020) Endocrine-disrupting chemicals and male reproductive health. Reprod Med Biol 19:243–253

    Google Scholar 

  • She J, Feng N, Zheng W, Zheng H, Cai P, Zou H, Yuan Y, Gu J, Liu Z, Bian J (2021) Zearalenone exposure disrupts blood–testis barrier integrity through excessive Ca2+-mediated autophagy. Toxins 13:875

    CAS  Google Scholar 

  • Shi Y, Vistro WA, Bai X, Wu R, Chen C, Huang Y, Fazlani SA, Tarique I, Yang P, Chen Q (2020) Effect of seasonal variance on intestinal epithelial barriers and the associated innate immune response of the small intestine of the Chinese soft-shelled turtles. Fish Shellfish Immunol 97:173–181

    CAS  Google Scholar 

  • Song W-H, Yi Y-J, Sutovsky M, Meyers S, Sutovsky P (2016) Autophagy and ubiquitin–proteasome system contribute to sperm mitophagy after mammalian fertilization. Proc Natl Acad Sci 113:E5261–E5270

    CAS  Google Scholar 

  • Sun J, Bian Y, Ma Y, Ali W, Wang T, Yuan Y, Gu J, Bian J, Liu Z, Zou H (2023) Melatonin alleviates cadmium-induced nonalcoholic fatty liver disease in ducks by alleviating autophagic flow arrest via PPAR-α and reducing oxidative stress. Poult Sci 102835

  • Sutovsky P, Moreno R, Ramalho-Santos J, Dominko T, Thompson WE, Schatten G (2001) A putative, ubiquitin-dependent mechanism for the recognition and elimination of defective spermatozoa in the mammalian epididymis. J Cell Sci 114:1665–1675

    CAS  Google Scholar 

  • Tarique I, Shi Y, Gandahi NS, Ding B, Yang P, Chen C, Vistro WA, Chen Q (2020) In vivo cellular evidence of autophagic associated spermiophagy within the principal cells during sperm storage in epididymis of the turtle. Aging (Albany NY) 12:8987

    CAS  Google Scholar 

  • Tian Y, Li Z, Hu W, Ren H, Tian E, Zhao Y, Lu Q, Huang X, Yang P, Li X (2010) C. elegans screen identifies autophagy genes specific to multicellular organisms. Cell 141:1042–1055

    CAS  Google Scholar 

  • Varga VB, Schuller D, Szikszai F, Szinyákovics J, Puska G, Vellai T, Kovács T (2022) Autophagy is required for spermatogonial differentiation in the Drosophila testis. Biologia Futura, pp 1–18

    Google Scholar 

  • Vistro WA, Zhang Y, Bai X, Yang P, Huang Y, Qu W, Baloch AS, Wu R, Tarique I, Chen Q (2019) In vivo autophagy up-regulation of small intestine enterocytes in chinese soft-shelled turtles during hibernation. Biomolecules 9:682

    Google Scholar 

  • Wang F, Han D (2019) Sertoli cell phagocytosis: an essential event for spermatogenesis. Male Reprod Health 11:126–319

  • Wang H, Wang H, Xiong W, Chen Y, Ma Q, Ma J, Ge Y, Han D (2006) Evaluation on the phagocytosis of apoptotic spermatogenic cells by Sertoli cells in vitro through detecting lipid droplet formation by Oil Red O staining. Reproduction 132:485–492

    CAS  Google Scholar 

  • Wang L, Zhang R, Wu B, Yu Y, Li W, Li S, Liu C (2022a) Autophagy mediated tubulobulbar complex components degradation is required for spermiation. Fundamental Research

    Google Scholar 

  • Wang M, Zeng L, Su P, Ma L, Zhang M, Yz Z (2022b) Autophagy: a multifaceted player in the fate of sperm. Hum Reprod Update 28:200–231

    CAS  Google Scholar 

  • Wang M-C, Wu A-G, Huang Y-Z, Shao G-L, Ji S-F, Wang R-W, Yuan H-J, Fan X-L, Zheng L-H, Jiao Q-L (2015) Autophagic regulation of cell growth by altered expression of Beclin 1 in triple-negative breast cancer. Int J Clin Exp Med 8:7049

    Google Scholar 

  • Wang T, Zhu Q, Cao B, Cai Y, Wen S, Bian J, Zou H, Song R, Gu J, Liu X (2022c) Ca2+ transfer via the ER-mitochondria tethering complex in neuronal cells contribute to cadmium-induced autophagy. Cell Biol Toxicol 38:469–485

    CAS  Google Scholar 

  • Wang Y, Yan J, Yin F, Li L, Qin Y, Meng C, Lu R, Guo L (2017) Role of autophagy in cadmium-induced testicular injury. Hum Exp Toxicol 36:1039–1048

    CAS  Google Scholar 

  • Yang P, Ahmed N, Wang L, Chen H, Waqas Y, Liu T, Haseeb A, Bangulzai N, Huang Y, Chen Q (2017) In vivo autophagy and biogenesis of autophagosomes within male haploid cells during spermiogenesis. Oncotarget 8:56791

    Google Scholar 

  • Yang S-H, He J-B, Yu L-H, Li L, Long M, Liu M-D, Li P (2019) Protective role of curcumin in cadmium-induced testicular injury in mice by attenuating oxidative stress via Nrf2/ARE pathway. Environ Sci Pollut Res 26:34575–34583

    CAS  Google Scholar 

  • Yao Y, Wan Y, Shi X, Guo L, Jiang H, Zhang X, Xu B, Hua J (2022) Letrozole protects against cadmium-induced inhibition of spermatogenesis via LHCGR and Hsd3b6 to activate testosterone synthesis in mice. Reprod Biol Endocrinol 20:43

    CAS  Google Scholar 

  • Yuan J, Zhao Y, Bai Y, Gu J, Yuan Y, Liu X, Liu Z, Zou H, Bian J (2021) Cadmium induces endosomal/lysosomal enlargement and blocks autophagy flux in rat hepatocytes by damaging microtubules. Ecotoxicol Environ Saf 228:112993

    CAS  Google Scholar 

  • Zhang J, Xiong Y-W, Tan L-L, Zheng X-M, Zhang Y-F, Ling Q, Zhang C, Zhu H-L, Chang W, Wang H (2023a) Sperm Rhoa m6A modification mediates intergenerational transmission of paternally acquired hippocampal neuronal senescence and cognitive deficits after combined exposure to environmental cadmium and high-fat diet in mice. J Hazard Mater 131891

  • Zhang L, Wang Y, Yi L, Huang D, Zhang Y, Mirqami KA, Cheng Y, Ren Q (2023b) Effect of autophagy in cadmium chloride induced apoptosis of mouse spermatogenic cells. Wei Sheng Yan Jiu= J Hyg Res 52:489–496

    Google Scholar 

  • Zhu Q, Li X, Ge R-S (2020) Toxicological effects of cadmium on mammalian testis. Front Genet 11:527

    CAS  Google Scholar 

  • Zhu Y, Yin Q, Wei D, Yang Z, Du Y, Ma Y (2019) Autophagy in male reproduction. Syst Biol Reprod Med 65:265–272

    Google Scholar 

  • Zou H, Sun J, Wu B, Yuan Y, Gu J, Bian J, Liu X, Liu Z (2020a) Effects of cadmium and/or lead on autophagy and liver injury in rats. Biol Trace Elem Res 198:206–215

    CAS  Google Scholar 

  • Zou H, Wang T, Yuan J, Sun J, Yuan Y, Gu J, Liu X, Bian J, Liu Z (2020b) Cadmium-induced cytotoxicity in mouse liver cells is associated with the disruption of autophagic flux via inhibiting the fusion of autophagosomes and lysosomes. Toxicol Lett 321:32–43

    CAS  Google Scholar 

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Funding

The National Natural Science Foundation (grant numbers 31702305, 31872533, and 32072933) and Jiangsu Higher Education Institutions Priority Academic Program Development (PAPD) supported this study.

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Authors and Affiliations

  1. College of Veterinary Medicine, Yangzhou University Yangzhou, Yangzhou, Jiangsu, 225009, People’s Republic of China

    Waseem Ali, Kai Deng, Jian Sun, Yonggang Ma, Zongping Liu & Hui Zou

  2. Joint International Research Laboratory of Agriculture and Agri-Product Safety of the Ministry of Education of China, Yangzhou University, Yangzhou, Jiangsu, 225009, People’s Republic of China

    Waseem Ali, Kai Deng, Jian Sun, Yonggang Ma, Zongping Liu & Hui Zou

  3. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu, 225009, People’s Republic of China

    Waseem Ali, Kai Deng, Jian Sun, Yonggang Ma, Zongping Liu & Hui Zou

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  1. Waseem Ali
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  2. Kai Deng
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  3. Jian Sun
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  4. Yonggang Ma
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Contributions

Zongping Liu and Hui Zou: data curation, original draft writing, review, and editing. Waseem Ali: original draft writing. Kai Deng and Yonggang Ma: validation and software. Jian Sun: investigation and visualization. Zongping Liu is responsible for conceptualization, methodology, and software development.

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Correspondence to Hui Zou.

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Ali, W., Deng, K., Sun, J. et al. A new insight of cadmium-induced cellular evidence of autophagic-associated spermiophagy during spermatogenesis. Environ Sci Pollut Res 30, 101064–101074 (2023). https://doi.org/10.1007/s11356-023-29548-9

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