Retrograde Analysis of Calcium Signaling by CaMPARI2 Shows Cytosolic Calcium in Chondrocytes Is Unaffected by Parabolic Flights

Calcium (Ca2+) elevation is an essential secondary messenger in many cellular processes, including disease progression and adaptation to external stimuli, e.g., gravitational load. Therefore, mapping and quantifying Ca2+ signaling with a high spatiotemporal resolution is a key challenge. However, particularly on microgravity platforms, experiment time is limited, allowing only a small number of replicates. Furthermore, experiment hardware is exposed to changes in gravity levels, causing experimental artifacts unless appropriately controlled. We introduce a new experimental setup based on the fluorescent Ca2+ reporter CaMPARI2, onboard LED arrays, and subsequent microscopic analysis on the ground. This setup allows for higher throughput and accuracy due to its retrograde nature. The excellent performance of CaMPARI2 was demonstrated with human chondrocytes during the 75th ESA parabolic flight campaign. CaMPARI2 revealed a strong Ca2+ response triggered by histamine but was not affected by the alternating gravitational load of a parabolic flight.

Motor imagery helps updating internal models during microgravity exposure

Skilled movements result from a mixture of feedforward and feedback mechanisms conceptualized by internal models. These mechanisms subserve both motor execution and motor imagery. Current research suggests that imagery allows updating feedforward mechanisms, leading to better performance in familiar contexts. Does this still hold in radically new contexts? Here, we test this ability by asking participants to imagine swinging arm movements around shoulder in normal gravity condition and in microgravity in which studies showed that movements slow down. We timed several cycles of actual and imagined arm pendular movements in three groups of subjects during parabolic flight campaign. The first, control, group remained on the ground. The second group was exposed to microgravity but did not imagine movements inflight. The third group was exposed to microgravity and imagined movements inflight. All groups performed and imagined the movements before and after the flight. We predicted that a mere exposure to microgravity would induce changes in imagined movement duration. We found this held true for the group who imagined the movements, suggesting an update of internal representations of gravity. However, we did not find a similar effect in the group exposed to microgravity despite the fact participants lived the same gravitational variations as the first group. Overall, these results suggest that motor imagery contributes to update internal representations of movement in unfamiliar environments, while a mere exposure proved to be insufficient.

Proteomic Response of Deinococcus radiodurans to Short-Term Real Microgravity during Parabolic Flight Reveals Altered Abundance of Proteins Involved in Stress Response and Cell Envelope Functions

Rapidly evolving space exploration makes understanding the short- and long- term effects of microgravity on humans, plants, and microorganisms an important task. The ubiquitous presence of the gravitational force has had an influence on the development of all living entities on Earth, and short- and long-term changes in perceived gravitational force can induce notable changes within cells. Deinococcus radiodurans is the Gram-positive bacterium that is best known for its extreme resistance to UV-C and gamma radiation, oxidation stress, and desiccation. Thus increased interest has been placed on this species in the context of space research. The present study aims to elucidate the short-term proteomic response of this species to real microgravity during parabolic flight. Overnight cultures of D. radiodurans were subjected to microgravity during a single parabola, and metabolic activity was quenched using methanol. Proteins were extracted and subsequently measured using HPLC nESI MS/MS. The results, such as the enrichment of the peptidoglycan biosynthesis pathway with differentially abundant proteins and altered S-layer protein abundance, suggested molecular rearrangements in the cell envelope of D. radiodurans. Altered abundance of proteins involved in energy metabolism and DNA repair could be linked with increased endogenous ROS production that contributes to the stress response. Moreover, changes in protein abundance in response to microgravity show similarities with previously reported stress responses. Thus, the present results could be used to further investigate the complex regulation of the remarkable stress management of this bacterium.

Effects of Simulated Microgravity and Hypergravity Conditions on Arm Movements in Normogravity

The human sensorimotor control has evolved in the Earth’s environment where all movement is influenced by the gravitational force. Changes in this environmental force can severely impact the performance of arm movements which can be detrimental in completing certain tasks such as piloting or controlling complex vehicles. For this reason, subjects that are required to perform such tasks undergo extensive training procedures in order to minimize the chances of failure. We investigated whether local gravity simulation of altered gravitational conditions on the arm would lead to changes in kinematic parameters comparable to the full-body experience of microgravity and hypergravity onboard a parabolic flight. To see if this would be a feasible approach for on-ground training of arm reaching movements in altered gravity conditions we developed a robotic device that was able to apply forces at the wrist in order to simulate micro- or hypergravity conditions for the arm while subjects performed pointing movements on a touch screen. We analyzed and compared the results of several kinematic parameters along with muscle activity using this system with data of the same subjects being fully exposed to microgravity and hypergravity conditions on a parabolic flight. Both in our simulation and in-flight, we observed a significant increase in movement durations in microgravity conditions and increased velocities in hypergravity for upward movements. Additionally, we noted a reduced accuracy of pointing both in-flight and in our simulation. These promising results suggest, that locally simulated altered gravity can elicit similar changes in some movement characteristics for arm reaching movements. This could potentially be exploited as a means of developing devices such as exoskeletons to aid in training individuals prior to undertaking tasks in changed gravitational conditions.

Fidelity of a Bacterial DNA Polymerase in Microgravity, a Model for Human Health in Space

Long-term space missions will expose crew members, their cells as well as their microbiomes to prolonged periods of microgravity and ionizing radiation, environmental stressors for which almost no earth-based organisms have evolved to survive. Despite the importance of maintaining genomic integrity, the impact of these stresses on DNA polymerase-mediated replication and repair has not been fully explored. DNA polymerase fidelity and replication rates were assayed under conditions of microgravity generated by parabolic flight and compared to earth-like gravity. Upon commencement of a parabolic arc, primed synthetic single-stranded DNA was used as a template for one of two enzymes (Klenow fragment exonuclease+/−; with and without proofreading exonuclease activity, respectively) and were quenched immediately following the 20 s microgravitational period. DNA polymerase error rates were determined with an algorithm developed to identify experimental mutations. In microgravity Klenow exonuclease+ showed a median 1.1-fold per-base decrease in polymerization fidelity for base substitutions when compared to earth-like gravity (p = 0.02), but in the absence of proofreading activity, a 2.4-fold decrease was observed (p = 1.98 × 10−11). Similarly, 1.1-fold and 1.5-fold increases in deletion frequencies in the presence or absence of exonuclease activity (p = 1.51 × 10−7 and p = 8.74 × 10−13), respectively, were observed in microgravity compared to controls. The development of this flexible semi-autonomous payload system coupled with genetic and bioinformatic approaches serves as a proof-of-concept for future space health research.

The effects of real and simulated microgravity on cellular mitochondrial function

Astronauts returning from space shuttle missions or the International Space Station have been diagnosed with various health problems such as bone demineralization, muscle atrophy, cardiovascular deconditioning, and vestibular and sensory imbalance including visual acuity, altered metabolic and nutritional status, and immune system dysregulation. These health issues are associated with oxidative stress caused by a microgravity environment. Mitochondria are a source of reactive oxygen species (ROS). However, the molecular mechanisms through which mitochondria produce ROS in a microgravity environment remain unclear. Therefore, this review aimed to explore the mechanism through which microgravity induces oxidative damage in mitochondria by evaluating the expression of genes and proteins, as well as relevant metabolic pathways. In general, microgravity-induced ROS reduce mitochondrial volume by mainly affecting the efficiency of the respiratory chain and metabolic pathways. The impaired respiratory chain is thought to generate ROS through premature electron leakage in the electron transport chain. The imbalance between ROS production and antioxidant defense in mitochondria is the main cause of mitochondrial stress and damage, which leads to mitochondrial dysfunction. Moreover, we discuss the effects of antioxidants against oxidative stress caused by the microgravity environment space microgravity in together with simulated microgravity (i.e., spaceflight or ground-based spaceflight analogs: parabolic flight, centrifugal force, drop towers, etc.). Further studies should be taken to explore the effects of microgravity on mitochondrial stress-related diseases, especially for the development of new therapeutic drugs that can help increase the health of astronauts on long space missions.

Cardiopulmonary responses to centrifuge simulated parabolic flight

Introduction: Parabolic flights, by producing short periods of weightlessness, closely simulate microgravity. However, they are still expensive, incur a significant logistics support, and occurrence of any adverse events during such simulation is undesirable. The present study was formulated to explore the feasibility of using a human centrifuge for simulation of parabolic flight to study the cardiopulmonary parameters as an alternative ground-based model. Material and Methods: Twelve healthy male volunteers were subjected to simulated parabolic flight, the profile of which involved exposure to 20 repetitions of hypogravity periods (+0.5 Gz), each interposed between periods of hypergravity phases (+2 Gz), using high-performance human centrifuge. Heart rate (HR), respiratory rate (RR), and arterial oxygen saturation (SpO2) were studied during such a simulation and analyzed using one-way repeated measures ANOVA. Motion sickness assessment questionnaire was administered to the participants after the run. They were also asked to rate their subjective feeling of weightlessness experienced during the run. Results: Comparison of HR revealed a significant difference (F = 22.167, P < 0.001) across 20 loops of different gravity phases. Post hoc analysis revealed that the mean HR of hypergravity phases was significantly higher compared with pre-run 1 G values and that of hypogravity phases. Similarly, HR showed a significant difference across pre-run 1 G, 10th and 20th loops of hypogravity phases (F = 5.672, P = 0.01). Post hoc analysis revealed a significant reduction in HR at 20th loop compared to both pre-run 1 G (P = 0.023) and 10th loop (P = 0.042) values. No significant differences were observed in both RR (F = 1.789, P = 0.148) and SpO2 (F = 1.708, P =0.199) across different gravity phases. The mean overall motion sickness score was found to be 23.6%. Participants rated their subjective feeling of weightlessness between 4 and 6 (mode = 5) on a scale of 1–10. Conclusion: It can be concluded from the results that HR increased during hypergravity conditions and reduced during hypogravity conditions, an expected outcome during parabolic flight. The significant reduction in HR during the 20th loop of hypogravity phase compared to 10th loop and pre-run 1 G conditions indicate a possible association with the duration of exposure. The centrifuge simulated parabolic flight profile designed in our study was able to emanate physiological changes similar to those experienced in actual parabolic flight for HR, RR, and SpO2.