The general objective of the study was to determine modulation of gene expression by environmental factors with a special emphasis on bone formation. For this reason the specific period of treatment was chosen between 5-6 days post-fertilization (dpf) when bone formation and calcification are taking place. Zebrafish larvae were placed at 5 dpf into a Large Diameter Centrifuge and brought to a gravitational force of 3g or 5g for 24 hours. We show that this treatment causes a clear increase of bone formation as illustrated by cranial skeleton staining of the bone matrix by Alizarin Red by morphometric analysis of the resulting images and by gene expression studies of selected genes. Thus a whole genome micro-array experiment was conducted to identify genes that may be involved in the observed effect on bone formation.

The general objective of the study was to determine modulation of gene expression by environmental factors with a special emphasis on bone formation. For this reason the specific period of treatment was chosen between 5-6 days post-fertilization (dpf) when bone formation and calcification are taking place. This experiment was designed as a new type of gravitational experiment which we like to call relative microgravity referring to the fact that the larvae first grow in hyper gravity for 5 days and are then returned to 1g normal gravity for 1 day. Zebrafish embryos were placed on a Large Diameter Centrifuge at 3 hpf brought to a gravitational force of 3 g until 5 dpf. Reference embryos were kept in parallel at 1g (Inc). At 5dpf one batch was left at 3g (3g) one batch was returned to 1g (3g>1g) while a third batch was returned to 1g but left on the axis of the centrifuge (Axe; 3g>Axe). The experiment was repeated 4 times each time with 4 batches of 60 larvae.

The general objective of the study was to determine modulation of gene expression by environmental factors with a special emphasis on bone formation. For this reason the specific period of treatment was chosen between 5-6 days post-fertilization (dpf) when bone formation and calcification are taking place. This experiment was designed as a new type of gravitational experiment which we like to call relative microgravity referring to the fact that the larvae first grow in hyper gravity for 5 days and are then returned to 1g normal gravity for 1 day. Zebrafish embryos were placed on a Large Diameter Centrifuge at 3 hpf brought to a gravitational force of 3 g until 5 dpf. Reference embryos were kept in parallel at 1g (Inc). At 5dpf one batch was left at 3g (3g) one batch was returned to 1g (3g>1g) while a third batch was returned to 1g but left on the axis of the centrifuge (Axe; 3g>Axe). The experiment was repeated 4 times each time with 4 batches of 60 larvae.

The general objective of the study was to determine modulation of gene expression by environmental factors specifically simulation of microgravity with a special emphasis on bone formation. For this reason the specific period of treatment was chosen between 5-6 days post-fertilization (dpf) when bone formation and calcification are taking place. Zebrafish larvae were placed at 5 dpf into a clinostat (CLINO) for 24 hours which was shown to simulate microgravity by the specific rotational movements it generates. We show that CLINO exposure causes a clear decrease of bone formation as illustrated by cranial skeleton staining of the bone matrix by Alizarin Red by morphometric analysis of the resulting images. Thus a whole genome micro-array experiment was conducted to identify genes that may be involved in the observed effect on bone formation.

The general objective of the study was to determine modulation of gene expression by environmental factors specifically simulation of microgravity with a special emphasis on bone formation. For this reason the specific period of treatment was chosen between 5-6 days post-fertilization (dpf) when bone formation and calcification are taking place. Zebrafish larvae were placed at 5 dpf into a clinostat (CLINO) for 24 hours which was shown to simulate microgravity by the specific rotational movements it generates. We show that CLINO exposure causes a clear decrease of bone formation as illustrated by cranial skeleton staining of the bone matrix by Alizarin Red by morphometric analysis of the resulting images. Thus a whole genome micro-array experiment was conducted to identify genes that may be involved in the observed effect on bone formation.

['To reveal the potential mechanisms involved in the dysfunction of antiviral immune responses under simulated microgravity conditions, we investigated the transcriptional changes related to the status of innate immune responses by RNA-seq with poly I:C or mock PBS treatment under Normal gravity or simulated microgravity conditions. Our results indicate that the retinoic acid inducible gene (RIG)-I-like receptor (RLR) and Toll-like receptor (TLR) signal pathways, which are both involved in the type-I interferon induction, are significantly inhibited by simulated microgravity effects.']

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/toxics10050256/s1. Figure S1, Effect of DMSO on Light/Dark Locomotor Activity; Figure S2, Time Course Behavioral Graph, Activity Box Plots and Developmental Toxicity for each Chemical; Table S1, Raw Data. This dataset is associated with the following publication: Jarema, K., D. Hunter, B. Hill, J. Olin, K. Britton, M. Waalkes, and S. Padilla. Developmental Neurotoxicity and Behavioral Screening in Larval Zebrafish with a Comparison to Other Published Results. Toxics. MDPI, Basel, SWITZERLAND, 10(5): 256, (2022).

Data used to generate figures shown in Phelps et al. This dataset is associated with the following publication: Phelps, D., N. Brinkman, S. Keely, E. Anneken, T. Catron, D. Betancourt, C. Wood, S. Espenschied, J. Rawls, and T. Tal. Microbial colonization is required for normal neurobehavioral development in zebrafish. Scientific Reports. Nature Publishing Group, London, UK, 11(7): 11244, (2017).

Larvae-Pupae transition flies (Drosophila) were recovered and transport for 3 days at 12-14C to arrest development until the launch site then exposed to RT (18-20C) for some hours including the launch and trip to the International Space Station then pupae were exposed to microgravity in the ISS for 4 days and a half at 22C. Finally pupae were fixed on acetone and frozen until recovery on Earth. Four groups of samples: 1 ISS (+ground control) as described 2 RPM (microgravity simulator on Earth) as described 3 RPM without constrains (No MAMBA container and only 5 days exposure without cold transport) and 4 centrifuge 10g without constrains control.

Supporting Information for "Gust KA, Mylroie JE, Kimble AN, Wilbanks MS, Steward CSC, Chapman KA, Jensen KM, Kennedy AJ, Krupa PM, Waisner SA, Pandelides Z, Garcia-Reyero N, Erickson RJ, Ankley GT, Conder J, Moore DW. Survival, Growth, and Reproduction Responses in a Three-Generation Exposure of the Zebrafish (Danio rerio) to Perfluorooctane Sulfonate. Environ Toxicol Chem. 2024 Jan;43(1):115-131. doi: 10.1002/etc.5770. Epub 2023 Nov 29. PMID: 38018867.". This dataset is associated with the following publication: Gust, K., J. Mylroie, A. Kimble, M. Wilbanks, C. Steward, K. Chapman, K. Jensen, A. Kennedy, P. Krupa, S. Waisner, Z. Pandelides, N. Garcia-Reyero, R. Erickson, G. Ankley, J. Conder, and D. Moore. Survival, Growth, and Reproduction Responses in a Three-Generation Exposure of the Zebrafish (Danio rerio) to Perfluorooctane Sulfonate. ENVIRONMENTAL TOXICOLOGY AND CHEMISTRY. Society of Environmental Toxicology and Chemistry, Pensacola, FL, USA, 43(1): 115-131, (2024).