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1.
NPJ Microgravity ; 10(1): 50, 2024 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-38693246

RESUMO

Periodically, the European Space Agency (ESA) updates scientific roadmaps in consultation with the scientific community. The ESA SciSpacE Science Community White Paper (SSCWP) 9, "Biology in Space and Analogue Environments", focusses in 5 main topic areas, aiming to address key community-identified knowledge gaps in Space Biology. Here we present one of the identified topic areas, which is also an unanswered question of life science research in Space: "How to Obtain an Integrated Picture of the Molecular Networks Involved in Adaptation to Microgravity in Different Biological Systems?" The manuscript reports the main gaps of knowledge which have been identified by the community in the above topic area as well as the approach the community indicates to address the gaps not yet bridged. Moreover, the relevance that these research activities might have for the space exploration programs and also for application in industrial and technological fields on Earth is briefly discussed.

2.
NPJ Microgravity ; 10(1): 16, 2024 Feb 10.
Artigo em Inglês | MEDLINE | ID: mdl-38341423

RESUMO

Progress in mechanobiology allowed us to better understand the important role of mechanical forces in the regulation of biological processes. Space research in the field of life sciences clearly showed that gravity plays a crucial role in biological processes. The space environment offers the unique opportunity to carry out experiments without gravity, helping us not only to understand the effects of gravitational alterations on biological systems but also the mechanisms underlying mechanoperception and cell/tissue response to mechanical and gravitational stresses. Despite the progress made so far, for future space exploration programs it is necessary to increase our knowledge on the mechanotransduction processes as well as on the molecular mechanisms underlying microgravity-induced cell and tissue alterations. This white paper reports the suggestions and recommendations of the SciSpacE Science Community for the elaboration of the section of the European Space Agency roadmap "Biology in Space and Analogue Environments" focusing on "How are cells and tissues influenced by gravity and what are the gravity perception mechanisms?" The knowledge gaps that prevent the Science Community from fully answering this question and the activities proposed to fill them are discussed.

3.
NPJ Microgravity ; 9(1): 84, 2023 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-37865644

RESUMO

The present white paper concerns the indications and recommendations of the SciSpacE Science Community to make progress in filling the gaps of knowledge that prevent us from answering the question: "How Do Gravity Alterations Affect Animal and Human Systems at a Cellular/Tissue Level?" This is one of the five major scientific issues of the ESA roadmap "Biology in Space and Analogue Environments". Despite the many studies conducted so far on spaceflight adaptation mechanisms and related pathophysiological alterations observed in astronauts, we are not yet able to elaborate a synthetic integrated model of the many changes occurring at different system and functional levels. Consequently, it is difficult to develop credible models for predicting long-term consequences of human adaptation to the space environment, as well as to implement medical support plans for long-term missions and a strategy for preventing the possible health risks due to prolonged exposure to spaceflight beyond the low Earth orbit (LEO). The research activities suggested by the scientific community have the aim to overcome these problems by striving to connect biological and physiological aspects in a more holistic view of space adaptation effects.

4.
Rev Sci Instrum ; 85(10): 101101, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25362364

RESUMO

As commercial space flights have become feasible and long-term extraterrestrial missions are planned, it is imperative that the impact of space travel and the space environment on human physiology be thoroughly characterized. Scrutinizing the effects of potentially detrimental factors such as ionizing radiation and microgravity at the cellular and tissue level demands adequate visualization technology. Advanced light microscopy (ALM) is the leading tool for non-destructive structural and functional investigation of static as well as dynamic biological systems. In recent years, technological developments and advances in photochemistry and genetic engineering have boosted all aspects of resolution, readout and throughput, rendering ALM ideally suited for biological space research. While various microscopy-based studies have addressed cellular response to space-related environmental stressors, biological endpoints have typically been determined only after the mission, leaving an experimental gap that is prone to bias results. An on-board, real-time microscopical monitoring device can bridge this gap. Breadboards and even fully operational microscope setups have been conceived, but they need to be rendered more compact and versatile. Most importantly, they must allow addressing the impact of gravity, or the lack thereof, on physiologically relevant biological systems in space and in ground-based simulations. In order to delineate the essential functionalities for such a system, we have reviewed the pending questions in space science, the relevant biological model systems, and the state-of-the art in ALM. Based on a rigorous trade-off, in which we recognize the relevance of multi-cellular systems and the cellular microenvironment, we propose a compact, but flexible concept for space-related cell biological research that is based on light sheet microscopy.


Assuntos
Biologia/métodos , Meio Ambiente Extraterreno , Microscopia/métodos , Pesquisa , Animais , Humanos , Imagem Molecular
5.
J Struct Biol ; 183(3): 455-466, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23792165

RESUMO

Combining the use of cells with sparse cristae marked with IMP-EGFP and short pulsed sub-saturating fluorescence excitation (non-saturation fluorescence microscopy/NSFM) revealed inhomogeneous fluorescence distribution along mitochondria in living cells. Also the matrix located TMRE was distributed non-uniformly and at least in part filling the gaps between the IMP-EGFP fluorescence: fluorescence intensities are modulated in space and time in part in an antidromic manner. The spatial modulations can be interpreted to represent cristae/matrix distributions. The temporal fluctuations of fluorescence vary within 0.3-3s. Because most peak positions of IMP fluorescence remain stationary up to at least several minutes, temporal intensity modulations may result from varying emissions related to the degree of excitation and/or represent wobbling of cristae, i.e. lateral movements, bending or size changes. Modulations by noise and non-saturated excitation have been reduced by 3 steps of deconvolution followed by averaging 4 images. This allowed a final temporal resolution of 150ms. Disappearance of cristae or formation of new ones takes place within a few seconds, but these are rare events. Thus position of cristae seems to be rather stable, but they regularly disassemble close to fission sites. Treatment with oligomycin strongly reduces "wobbling" activity.


Assuntos
Células Endoteliais da Veia Umbilical Humana/ultraestrutura , Membranas Mitocondriais/ultraestrutura , Corantes Fluorescentes/química , Corantes Fluorescentes/metabolismo , Proteínas de Fluorescência Verde/metabolismo , Células HeLa , Humanos , Microscopia de Fluorescência , Dinâmica Mitocondrial , Membranas Mitocondriais/efeitos dos fármacos , Oligomicinas/farmacologia , Forma das Organelas , Compostos Organometálicos/química , Compostos Organometálicos/metabolismo
6.
Scanning ; 31(3): 102-6, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-19444860

RESUMO

G3S1 cells are a new line derived from EM-G3 breast cancer cells by chronic nutritional stress and treatments with 12-O-tetradecanoylphorbol-13-acetate. These cells are capable of growing in standard medium. G3S1 cells exhibited elevated invasiveness in Matrigel invasion chambers as compared with parental EM-G3 cells. Elevated invasiveness of G3S1 cells was accompanied by higher incidence of myzitiras morphotype (sucker-like) and newly observed vthela morphotype (leech-like) both inducible in Hanks' Balanced Salt Solution test. Time-lapse phase contrast microscopy showed a capacity of G3S1 cells to form lobopodial protrusions already 20 min after seeding on gelatin. These protrusions could make contact with the dish and possibly produce the vthela shape. The possible relationship of mysitiras and vthela morphotypes to an increase in malignant potential marked by enhanced invasiveness was thus indicated.


Assuntos
Neoplasias da Mama/patologia , Animais , Linhagem Celular Tumoral , Feminino , Humanos , Camundongos , Microscopia Confocal , Microscopia de Contraste de Fase , Microscopia de Vídeo
7.
Mol Membr Biol ; 23(6): 509-20, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-17127623

RESUMO

Highly dynamic mitochondrial morphology is a prerequisite for fusion and fission. Mitochondrial fusion may represent a rescue mechanism for impaired mitochondria by exchanging constituents (proteins, lipids and mitochondrial DNA) and thus maintaining functionality. Here we followed for the first time the dynamics of a protein complex of the respiratory chain during fusion and fission. HeLa cells with differently labelled respiratory Complex I were fused and the dynamics of Complex I were investigated. The mitochondrial proteins spread throughout the whole mitochondrial population within 3 to 6 h after induction of cell fusion. Mitochondria of fused cells displayed a patchy substructure where the differently labelled proteins occupied separate and distinct spaces. This patchy appearance was already--although less pronounced--observed within single mitochondria before fusion, indicating a specific localization of Complex I with restricted diffusion within the inner membrane. These findings substantiate the view of a homogenous mitochondrial population due to constantly rearranging mitochondria, but also indicate the existence of distinct inner mitochondrial sub-compartments for respiratory chain complexes.


Assuntos
Complexo I de Transporte de Elétrons/metabolismo , Mitocôndrias/metabolismo , Membranas Mitocondriais/metabolismo , Animais , Células Cultivadas , Fluorescência , Células HeLa , Humanos , Fusão de Membrana , Mitocôndrias/fisiologia , Mitocôndrias/ultraestrutura , Transfecção , Xenopus laevis/metabolismo
8.
J Cell Sci ; 118(Pt 11): 2435-40, 2005 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-15923656

RESUMO

Tight regulation of the cell volume is important for the maintenance of cellular homeostasis. In a hypotonic environment, cells swell owing to osmosis. With many vertebrate cells, swelling is followed by an active reduction of volume, a process called regulatory volume decrease (RVD). A possible participant in RVD is the non-selective cation channel TRPV4, a member of the TRP superfamily that has been shown to react to hypotonic stimuli with a conductance for Ca2+. As a model for cell-volume regulation, we used a human keratinocyte cell line (HaCaT) that produces TRPV4 endogenously. When HaCaT cells were exposed to a hypotonic solution (200 mOsm) maximal swelling was followed by RVD. During swelling and volume regulation, a strong Ca2+ influx was measured. Gd3+, an inhibitor of TRPV4, blocked RVD of HaCaT cells and the accompanying rise of cytosolic Ca2+. To define the role of TRPV4 in volume regulation, a TRPV4-EGFP fusion protein was produced in CHO cells. CHO cells are unable to undergo RVD under hypotonic conditions and do not produce TRPV4 endogenously. Fluorescence imaging revealed that recombinant TRPV4 was localized to the cell membrane. Production of TRPV4 enabled CHO cells to undergo typical RVD after hypo-osmolarity-induced cell swelling. RVD of TRPV4-transfected CHO cells was significantly reduced by Gd3+ treatment or in Ca2+-free solution. Taken together, these results show a direct participation of TRPV4 in RVD.


Assuntos
Proteínas de Transporte de Cátions/metabolismo , Tamanho Celular , Canais Iônicos/metabolismo , Animais , Células CHO , Cálcio/metabolismo , Proteínas de Transporte de Cátions/antagonistas & inibidores , Proteínas de Transporte de Cátions/genética , Tamanho Celular/efeitos dos fármacos , Cricetinae , Citosol/metabolismo , Gadolínio/farmacologia , Expressão Gênica , Humanos , Soluções Hipotônicas , Canais Iônicos/antagonistas & inibidores , Canais Iônicos/genética , Osmose , Canais de Cátion TRPV
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