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See detailEffects of microgravity simulation on zebrafish transcriptomes and bone physiology; exposure starting at 5 days post-fertilization.
Aceto, Jessica ULg; Nourizadeh-Lillabadi, Rasoul; Bradamante, Silvia et al

in NPJ Microgravity (2016), 2

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See detailHormones, Simulated Microgravity and Hypergravity affect Bone and other Physiological Systems in Zebrafish Larvae
Aceto, Jessica ULg

Doctoral thesis (2015)

Zebrafish (Danio rerio) are increasingly used for physiological, genetic and developmental studies. Understanding the consequences of altered gravity on bone development and on general physiology in an ... [more ▼]

Zebrafish (Danio rerio) are increasingly used for physiological, genetic and developmental studies. Understanding the consequences of altered gravity on bone development and on general physiology in an entire organism remains to date incomplete. We used altered drug treatment and gravity experiments to evaluate their effects specifically on bone formation and more generally on whole genome gene expression. We started treatments at 5 days post-fertilization (dpf) and analyze early modifications in gene expression after 1 day using microarrays and the consequences on bone formation after 5 days using specific staining. We performed chemical treatments (Parathyroid Hormone, Vitamin D3), exposure to three different microgravity simulation devices (Clinostat, Random Positioning Machine and Rotating Wall Vessel) and finally exposure to hypergravity and "relative microgravity" in the Large Diameter Centrifuge. By combining morphometric tools with an objective scoring system for the state of development for each element in the head skeleton, and specific gene expression analysis, we confirmed and characterized in detail the decrease or increase of bone formation caused by a 5 day treatment (from 5dpf to 10dpf) with, respectively parathyroid hormone (PTH) or vitamin D3 (VitD3). Microarray transcriptome analysis after 24 hours treatment reveals a general effect on physiology upon VitD3 treatment, while PTH causes more specifically developmental effects. The microgravity simulators used were the 2D clinostat, random positioning machine and rotating wall vessel. Only clinorotation caused a significant decrease of bone formation when applied between 5 to 10dpf. This effect was not due to stress, as assessed by measuring cortisol levels in treated larvae. The two other devices caused no effect, or a slight acceleration of ossification. Gene expression results after one day in simulated microgravity indicate that musculo-skeletal, cardiavascular, and nuclear receptor systems are affected, however often in opposite directions in clinorotation compared to the two other devices. Based on the effects on bone formation and on the biological functions found to be affected, we conclude that clinorotation is the most appropriate method to simulate microgravity on ground when using free-swimming organisms such as zebrafish larvae. Hypergravity (3g from 5dpf to 9 dpf) exposure results in a significantly larger head and a significant increase in bone formation for a subset of the cranial bones. Gene expression analysis after 24hrs at 3g revealed differential expression of genes involved in the development and function of the skeletal, muscular, nervous, endocrine and cardiovascular systems. Finally, we propose a novel type of experimental approach, the "Reduced Gravity Paradigm", by keeping the developing larvae at 3g hypergravity for the first 5 days before returning them to 1g for one additional day. 5 days exposure to 3g during these early stages also caused increased bone formation, while gene expression analysis revealed a central network of regulatory genes (hes5, sox10, lgals3bp, egr1, edn1, fos, fosb, klf2, gadd45ba and socs3a) whose expression was consistently affected by the transition from hyper- to normal gravity. [less ▲]

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See detailZebrafish bone and general physiology are differently affected by hormones or changes in gravity.
Aceto, Jessica ULg; Nourizadeh-Lillabadi, Rasoul; Marée, Raphaël ULg et al

in PLoS ONE (2015), 10(6), 1-42

Teleost fish such as zebrafish (Danio rerio) are increasingly used for physiological, genetic and developmental studies. Our understanding of the physiological consequences of altered gravity in an entire ... [more ▼]

Teleost fish such as zebrafish (Danio rerio) are increasingly used for physiological, genetic and developmental studies. Our understanding of the physiological consequences of altered gravity in an entire organism is still incomplete. We used altered gravity and drug treatment experiments to evaluate their effects specifically on bone formation and more generally on whole genome gene expression. By combining morphometric tools with an objective scoring system for the state of development for each element in the head skeleton and specific gene expression analysis, we confirmed and characterized in detail the decrease or increase of bone formation caused by a 5 day treatment (from 5dpf to 10 dpf) of, respectively parathyroid hormone (PTH) or vitamin D3 (VitD3). Microarray transcriptome analysis after 24 hours treatment reveals a general effect on physiology upon VitD3 treatment, while PTH causes more specifically developmental effects. Hypergravity (3g from 5dpf to 9 dpf) exposure results in a significantly larger head and a significant increase in bone formation for a subset of the cranial bones. Gene expression analysis after 24 hrs at 3g revealed differential expression of genes involved in the development and function of the skeletal, muscular, nervous, endocrine and cardiovascular systems. Finally, we propose a novel type of experimental approach, the "Reduced Gravity Paradigm", by keeping the developing larvae at 3g hypergravity for the first 5 days before returning them to 1g for one additional day. 5 days exposure to 3g during these early stages also caused increased bone formation, while gene expression analysis revealed a central network of regulatory genes (hes5, sox10, lgals3bp, egr1, edn1, fos, fosb, klf2, gadd45ba and socs3a) whose expression was consistently affected by the transition from hyper- to normal gravity. [less ▲]

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See detailZebrafish Skeleton Measurements using Image Analysis and Machine Learning Methods
Stern, Olivier ULg; Marée, Raphaël ULg; Aceto, Jessica ULg et al

Poster (2011, May 20)

The zebrafish is a model organism for biological studies on development and gene function. Our work aims at automating the detection of the cartilage skeleton and measuring several distances and angles to ... [more ▼]

The zebrafish is a model organism for biological studies on development and gene function. Our work aims at automating the detection of the cartilage skeleton and measuring several distances and angles to quantify its development following different experimental conditions. [less ▲]

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See detailAutomatic localization of interest points in zebrafish images with tree-based methods
Stern, Olivier ULg; Marée, Raphaël ULg; Aceto, Jessica ULg et al

in Proceedings of the 6th IAPR International Conference on Pattern Recognition in Bioinformatics (2011)

In many biological studies, scientists assess effects of experimental conditions by visual inspection of microscopy images. They are able to observe whether a protein is expressed or not, if cells are ... [more ▼]

In many biological studies, scientists assess effects of experimental conditions by visual inspection of microscopy images. They are able to observe whether a protein is expressed or not, if cells are going through normal cell cycles, how organisms evolve in different experimental conditions, etc. But, with the large number of images acquired in high-throughput experiments, this manual inspection becomes lengthy, tedious and error-prone. In this paper, we propose to automatically detect specific interest points in microscopy images using machine learning methods with the aim of performing automatic morphometric measurements in the context of Zebrafish studies. We systematically evaluate variants of ensembles of classification and regression trees on four datasets corresponding to different imaging modalities and experimental conditions. Our results show that all variants are effective, with a slight advantage for multiple output methods, which are more robust to parameter choices. [less ▲]

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See detailAreas of Research:Life sciences:Animal Physiology
Horn, Eberhard; van Loon, Jack; Aceto, Jessica ULg et al

in Beyens, Daniel; Carotenuto, Luigi; van Loon, Jack (Eds.) et al Laboratory Science with Space Data: Accessing and using space-experiment data (2011)

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See detailTHE FUNCTION OF HMG-BOX TRANSCRIPTION FACTORS Sox4a AND Sox4b IN ZEBRAFISH BONE DEVELOPMENT AND HOMEOSTASIS.
Aceto, Jessica ULg; Motte, Patrick ULg; Martial, Joseph ULg et al

in Journal of Gravitational Physiology : A Journal of the International Society for Gravitational Physiology (2008), 15

In mammals, the Sox4 gene is involved in development of endocardial crests, the brain, the lung, teeth, gonads and lymphocytes. Recently, Sox4 was shown to control bone mass and mineralization in mice. In ... [more ▼]

In mammals, the Sox4 gene is involved in development of endocardial crests, the brain, the lung, teeth, gonads and lymphocytes. Recently, Sox4 was shown to control bone mass and mineralization in mice. In zebrafish, two homologs for the mammalian Sox4 are present, sox4a and sox4b. Here we investigate the function of the sox4a and sox4b genes in cartilage and bone development in zebrafish. Therefore, we focus our attention on the first bone structures to be formed, the head skeleton and more precisely the pharyngeal cartilage. We show that both genes are expressed in the pharyngeal region, albeit at different time points during development. Double in situ hybridization experiments are used to exactly define the particular tissues where they are expressed. Furthermore, microinjection experiments of antisense oligonucleotides are used to block translation of these specific genes and to define their precise function during cartilage and bone development. [less ▲]

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