https://doi.org/10.4081/jbr.2021.9875
Morphofunctional, viability and antioxidant system alterations on rat primary testicular cells exposed to simulated microgravity
Submitted: May 24, 2021
Accepted: October 16, 2021
Published: November 11, 2021
Accepted: October 16, 2021
Abstract Views: 1515
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All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.
Authors
This study focused on effects induced by Short-term Simulated Microgravity (SMG) condition on primary cell culture from pre-pubertal Wistar rats testis. Cells were analyzed for cytoskeletal and Sex Hormone Binding Globulin (SHBG/ABP) changes by immunofluorescence technique, for antioxidant system exploiting RT-PCR and cell viability. Cells were cultured for 6 and 24h on a three-dimensional clinostat, Random Positioning Machine (RPM). At the end of each experiment, once stopped the RPM rotation, cells were either fixed in paraformaldehyde or lysed and RNA extracted. In cells exposed to SMG the cytoskeleton became disorganized, microtubules fragmented and SHBG was already undetectable after 6h of treatment. Moreover, various antioxidant systems significantly increased after 24h of SMG exposure. Initially, SMG seemed to disturb antioxidant protection strategies allowing the testes to support sperm production, thus generating an aging-like state of oxidative stress. Studies on changes induced by short-term altered gravity conditions, carried out in real microgravity, could give more information on steroidogenesis and germ cell differentiation within the testis exposed to this condition and confirm the validity of simulation approach.
Mishra B, Luderer U. Reproductive hazards of space travel in women and men. Nat Rev Endocrinol 2019;15:713–30.
DOI: https://doi.org/10.1038/s41574-019-0267-6
Tou J, Ronca A, Grindeland R, Wade C. Models to study gravitational biology of mammalian reproduction. Biol Reprod 2002;67:1681–87.
DOI: https://doi.org/10.1095/biolreprod.102.007252
Zhu H, Wang H, Liu Z. Effects of real and simulated weightlessness on the cardiac and peripheral vascular functions of humans: a review. Int J Occup Med Environ Health 2015;28:793–802.
DOI: https://doi.org/10.13075/ijomeh.1896.00301
Tanaka K, Nishimura N, Kawai Y. Adaptation to microgravity deconditioning and countermeasures. J Physiol Sci 2017;67:271–81.
DOI: https://doi.org/10.1007/s12576-016-0514-8
Sominsky L, Hodgson DM, McLaughlin EA, et al. Linking stress and infertility: a novel role for ghrelin. Endocr Rev 2017;38:432–67.
DOI: https://doi.org/10.1210/er.2016-1133
Nargund VH. Effects of psychological stress on male fertility. Nat Rev Urol 2015;12:373–82.
DOI: https://doi.org/10.1038/nrurol.2015.112
Herranz R, Anken R, Boonstra J, et al. Ground-based facilities for simulation of microgravity: organism-specific recommendations for their use and recommended terminology. Astrobiology 2013;13:1–17.
DOI: https://doi.org/10.1089/ast.2012.0876
Grindeland RE, Popova IA, Vasques M, Arnaud SB. Cosmos 1887 mission overview: effects of microgravity on rat body and adrenal weights and plasma constituents. FASEB J 1990;4:105–9.
DOI: https://doi.org/10.1096/fasebj.4.1.2295371
Hadley JA, Hall JC, O'Brien A, Ball R. Effects of a simulated microgravity model on cell structure and function in rat testis and epididymis. J Appl Physiol 1992;72:748–59
DOI: https://doi.org/10.1152/jappl.1992.72.2.748
Vasques M, Lang C, Grindeland RE, et al. Comparison of hyper- and microgravity on rat muscle organ weights and selected plasma constituents. Aerosp Med Hum Perform 1998;69:2–8.
Strollo F, Barger L, Fuller C. Testosterone urinary excretion rate increases during hypergravity in male monkeys. J Gravit Physiol 2000;7:181–82.
Masini MA, Albi E, Barmo C, et al. Impact of long-term exposure to space environment on adult mammalian organisms: a study on mouse thyroid and testis. Plos one 2012;7:e35418.
DOI: https://doi.org/10.1371/journal.pone.0035418
Smith SM, Heer M, Wang Z, et al. Long-duration space flight and bed rest effect on testosterone and other steroids. J Clin Endocrinol Metab 2012;97:270–78.
DOI: https://doi.org/10.1210/jc.2011-2233
Uva BM, Masini MA, Sturla M, et al. Clinorotation-induced weightlessness influences the cytoskeleton of glial cells in culture. Brain Res 2002;934:132–39.
DOI: https://doi.org/10.1016/S0006-8993(02)02415-0
Sciola L, Cogoli-Greuter M, Cogoli A, et al. Influence of microgravity on mitogen binding and cytoskeleton in Jurkat cells. Adv Space Res 1999;24:801–5.
DOI: https://doi.org/10.1016/S0273-1177(99)00078-2
Infanger M, Kossmehl P, Shakibaei M, et al. Simulated weightlessness changes the cytoskeleton and extracellular matrix proteins in papillary thyroid carcinoma cells. Cell Tissue Res 2006;324:267–77.
DOI: https://doi.org/10.1007/s00441-005-0142-8
Tauber S, Hauschild S, Paulsen K, et al. Signal transduction in primary human T lymphocytes in altered gravity during parabolic flight and clinostat experiments. Cell Physiol Biochem 2015;35:1034–51.
DOI: https://doi.org/10.1159/000373930
Gmunder FH, Kiess M, Sonnefeld G, et al. A ground-based model to study the effects of weightlessness in lymphocytes. Biol Cell 1990;70:33–38.
DOI: https://doi.org/10.1016/0248-4900(90)90358-A
Uva BM, Strollo F, Ricci F, et al. Effect of conditions of three dimensional clinostating on testicular cell machinery. Acta astronautica 2007;60:391–96.
DOI: https://doi.org/10.1016/j.actaastro.2006.09.007
Masini M, Prato P. Effects of a simulated microgravity model on cell structure and function in mouse testis. J Biol Res 2010;83:29–32.
DOI: https://doi.org/10.4081/jbr.2010.4437
Aleshcheva G, Sahana J, Ma X, et al. Changes in morphology, gene expression and protein content in chondrocytes cultured on a random positioning machine. Plos One 2013;8:e79057.
DOI: https://doi.org/10.1371/journal.pone.0079057
Van Borst AG, van Loon JJWA. Technology and development for the random positioning machine, rpm. Micorgravity Sci Technol 2009;21:287.
DOI: https://doi.org/10.1007/s12217-008-9043-2
Coons AH, Leduc EH, Connolly JM. Studies on antibody I A method for the histochemical demonstration of specific antibody and its application to a study of the hyperimmune rabbit. J Exper Med 1955;102:49–59.
DOI: https://doi.org/10.1084/jem.102.1.49
Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction. Anal Biochem 1978;162:156–59.
DOI: https://doi.org/10.1016/0003-2697(87)90021-2
Pfaffl MW, Horgan GW, Dempfle L. Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res 2002;30-6.
DOI: https://doi.org/10.1093/nar/30.9.e36
Vandesompele J, De Preter K, Pattyn F, et al. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control. Genome Biol 2002;3:1–12.
DOI: https://doi.org/10.1186/gb-2002-3-7-research0034
Morabito C, Guarnieri S, Catizone A, et al. Transient increases in intracellular calcium and reactive oxygen species levels in TCam-2 cells exposed to microgravity. Sci Rep 2017; 7:148-56.
DOI: https://doi.org/10.1038/s41598-017-15935-z
Aitken RJ, Roman SD. Antioxidant systems and oxidative stress in the testes. Oxid Med Cell Longev 2008;1:15–24.
DOI: https://doi.org/10.4161/oxim.1.1.6843
Elkholi R, Chipuk JE. How do I kill thee? Let me count the ways: p53 regulates PARP‐1 dependent necrosis. Bio Essays 2014;36:46-51.
DOI: https://doi.org/10.1002/bies.201300117
Venditti P, Di Meo S. The role of reactive oxygen species in the life cycle of the mitochondrion. Int J Mol Sci 2020;21:21-73.
DOI: https://doi.org/10.3390/ijms21062173
How to Cite
Bonetto, V., Scarabelli, L., & Masini, M. A. (2021). Morphofunctional, viability and antioxidant system alterations on rat primary testicular cells exposed to simulated microgravity. Journal of Biological Research - Bollettino Della Società Italiana Di Biologia Sperimentale, 94(2). https://doi.org/10.4081/jbr.2021.9875
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