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Microgravity Exposure Alterations of Cellular Junctions Proteins in TCam-2 Cells: Localization and Interaction †

Department of Anatomy, Histology, Forensic-Medicine and Orthopedics, Section of Histology and Embryology, “Sapienza” University of Rome, 00161 Rome, Italy
Human Spaceflight and Scientific Research Unit, Italian Space Agency, 00161 Rome, Italy
Department of Neuroscience, Imaging and Clinical Sciences-CAST, “G. d’Annunzio” University of Chieti-Pescara, 66100 Chieti, Italy
Department of Experimental Medicine, Università degli Studi della Campania “Luigi Vanvitelli”, 80138 Naples, Italy
Author to whom correspondence should be addressed.
Presented at Cells, Cells and Nothing but Cells: Discoveries, Challenges and Directions, 6–8 March 2023; Available online:
Biol. Life Sci. Forum 2023, 21(1), 2;
Published: 16 March 2023


One of the most important hazards of the space environment is microgravity, which causes an alteration in the physiology of different systems, including the reproductive one. It is widely accepted that cytoskeleton is the microgravity-sensitive apparatus of the cells, and that cytoskeletal modifications are responsible for microgravity-triggered cell alterations. We established a 3D free-floating culture system from TCam-2 cell, a human seminoma cell line, and then exposed the obtained TCam-2 spheroids for 24 h at unitary gravity (UG), or under a simulated microgravity condition (SM), using the random position machine (RPM). We tested the cytoskeletal and junctional features of these samples using Western blot and confocal microscopy analysis to elucidate the impact of microgravity on the adherent and occluding junctions of TCam-2 spheroids. The junctional ultrastructure was studied using transmission electron microscopy (TEM). TEM analysis revealed the presence of occluding junctions both in UG or SM samples. Even if Western blot revealed no quantitative difference in actin and occludin proteins both in UG and SM exposed samples, fluorescence colocalization analysis showed a significative increase in the colocalization area of occludin and actin proteins in the superficial layer of TCam-2 spheroids grown in RPM conditions. This result let us speculate that tight junction functionality is different in UG and SM exposed spheroids. As far as adherent junctions are concerned, TEM analysis revealed adherent junctions both in UG or SM samples. Moreover, we observed by Western blot a trend in terms of the increase in the vimentin expression in SM exposed spheroids. Confocal microscopy analyses confirmed this significant increase. All together, these data suggest that simulated microgravity conditions in TCam-2 spheroids alter the tight junction assembly, while the increase in the intermediate filament’s structures can in part be associated with an enrichment in the adherent junctions. A functional investigation is needed to more deeply clarify this hypothesis.

Author Contributions

Conceptualization, A.C., G.R., L.G. and M.B.; methodology, M.B., L.G., M.A.M., C.M. and S.G.; software, A.C., G.R., M.B. and L.G.; validation, A.C., G.R., M.B. and L.G.; formal analysis, A.C., G.R., M.B. and L.G.; investigation, A.C., G.R., M.B. and L.G.; resources, A.C., G.R., M.B. and L.G.; data curation, A.C., G.R., M.B. and L.G.; writing—original draft preparation, A.C. and G.R.; writing—review and editing, M.B., L.G., M.A.M., C.M. and S.G.; visualization, A.C. and G.R.; supervision, F.F.; project administration, A.C. and F.F.; funding acquisition, A.C. and G.R. All authors have read and agreed to the published version of the manuscript.


This research was funded by grants from ASI–Italian Space Agency, number 2020-24-HH.0.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Not applicable.

Conflicts of Interest

The authors declare no conflict of interest.
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MDPI and ACS Style

Berardini, M.; Gesualdi, L.; Ferranti, F.; Mariggiò, M.A.; Morabito, C.; Guarnieri, S.; Ricci, G.; Catizone, A. Microgravity Exposure Alterations of Cellular Junctions Proteins in TCam-2 Cells: Localization and Interaction. Biol. Life Sci. Forum 2023, 21, 2.

AMA Style

Berardini M, Gesualdi L, Ferranti F, Mariggiò MA, Morabito C, Guarnieri S, Ricci G, Catizone A. Microgravity Exposure Alterations of Cellular Junctions Proteins in TCam-2 Cells: Localization and Interaction. Biology and Life Sciences Forum. 2023; 21(1):2.

Chicago/Turabian Style

Berardini, Marika, Luisa Gesualdi, Francesca Ferranti, Maria Addolorata Mariggiò, Caterina Morabito, Simone Guarnieri, Giulia Ricci, and Angela Catizone. 2023. "Microgravity Exposure Alterations of Cellular Junctions Proteins in TCam-2 Cells: Localization and Interaction" Biology and Life Sciences Forum 21, no. 1: 2.

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