Cytoskeleton, a mechano-sensing apparatus of the cells, is sensitive to gravity conditions1. In our previous study, we investigated the effects of simulated microgravity (SM) on the cytoskeleton of human male germ cells (TCam-2). We observed that SM for 24 h altered microtubule and actin organization2. To determine whether cytoskeletal changes occur during the early stages of SM exposure, we analysed the effects after a short 3 h period. Quantitative analyses were performed using Reverse Phase Protein Array (RPPA). The distribution pattern of cytoskeletal filaments was studied by indirect immunofluorescence (IIF) and Confocal microscopy, using maximum projections obtained from 1μm optical sections. RPPA analyses showed an increase in total cofilin and tubulin as early as 3h of exposure. Cellular cofilin exists in two forms: cofilin and phospho-cofilin (the latter represents the inactive form), both regulating actin polymerization and dynamics in the cytoplasm. However, both forms are also present in the nucleus, where they transiently influence gene regulation in response to specific stimuli. This dual localization highlights the dynamic regulatory role of cofilin phosphorylation in cellular processes3. Indirect IIF showed that, in SM samples, total cofilin levels increased while nuclear phospho-cofilin levels decreased. Consistent with RPPA data, tubulin levels also increased according to IIF. These observations suggest a possible perturbation of actin cytoskeleton caused by SM exposure, which in turn also affects tubulin polymerization and organization confirming the already known interaction between microtubules and microfilaments4. Future studies are necessary to better understand the biological significance of these modifications. Nonetheless, these results indicate that cytoskeleton remodelling occurs in TCam-2 cells as early as 3 h after SM exposure, providing further insight into how TCam-2 cells adapt to SM condition.
Microgravity-induced changes in cytoskeleton structure of TCam-2 cells. 17th International Congress of Histochemistry and Cytochemistry / Berardini, M., Gesualdi, L., Di Pauli, A., Signore, M., Broccolucci, S., Ferranti, F., Bizzarri, M., Tafani, M., Ricci, G., Catizone, A.. - (2025). (17th International Congress of Histochemistry and Cytochemistry (ICHC) Rimini ).
Microgravity-induced changes in cytoskeleton structure of TCam-2 cells. 17th International Congress of Histochemistry and Cytochemistry
M. Berardini;L. Gesualdi;A. Di Pauli;M. Bizzarri;M. Tafani;A. Catizone
2025
Abstract
Cytoskeleton, a mechano-sensing apparatus of the cells, is sensitive to gravity conditions1. In our previous study, we investigated the effects of simulated microgravity (SM) on the cytoskeleton of human male germ cells (TCam-2). We observed that SM for 24 h altered microtubule and actin organization2. To determine whether cytoskeletal changes occur during the early stages of SM exposure, we analysed the effects after a short 3 h period. Quantitative analyses were performed using Reverse Phase Protein Array (RPPA). The distribution pattern of cytoskeletal filaments was studied by indirect immunofluorescence (IIF) and Confocal microscopy, using maximum projections obtained from 1μm optical sections. RPPA analyses showed an increase in total cofilin and tubulin as early as 3h of exposure. Cellular cofilin exists in two forms: cofilin and phospho-cofilin (the latter represents the inactive form), both regulating actin polymerization and dynamics in the cytoplasm. However, both forms are also present in the nucleus, where they transiently influence gene regulation in response to specific stimuli. This dual localization highlights the dynamic regulatory role of cofilin phosphorylation in cellular processes3. Indirect IIF showed that, in SM samples, total cofilin levels increased while nuclear phospho-cofilin levels decreased. Consistent with RPPA data, tubulin levels also increased according to IIF. These observations suggest a possible perturbation of actin cytoskeleton caused by SM exposure, which in turn also affects tubulin polymerization and organization confirming the already known interaction between microtubules and microfilaments4. Future studies are necessary to better understand the biological significance of these modifications. Nonetheless, these results indicate that cytoskeleton remodelling occurs in TCam-2 cells as early as 3 h after SM exposure, providing further insight into how TCam-2 cells adapt to SM condition.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
