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Zebrafish Bone and General Physiology Are Differently Affected by Hormones or Changes in Gravity.

Aceto J, Nourizadeh-Lillabadi R, Marée R, Dardenne N, Jeanray N, Wehenkel L, Aleström P, van Loon JJ, Muller M - PLoS ONE (2015)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Laboratory for Organogenesis and Regeneration, GIGA- Research, University of Liège, B-4000, Liège, Sart-Tilman, Belgium.

ABSTRACT
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.

No MeSH data available.


Related in: MedlinePlus

Network of genes affected in "relative microgravity" experiments.A network was constructed using the genes common to all three experiments, or the genes common only to 3g>1g and 3g>axe. Color overlay indicates the fold change relative to the 3g sample taken as control. Genes up-regulated (red), down-regulated (green), (*) indicates that the gene is represented by two or more probes on the microarray.
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pone.0126928.g011: Network of genes affected in "relative microgravity" experiments.A network was constructed using the genes common to all three experiments, or the genes common only to 3g>1g and 3g>axe. Color overlay indicates the fold change relative to the 3g sample taken as control. Genes up-regulated (red), down-regulated (green), (*) indicates that the gene is represented by two or more probes on the microarray.

Mentions: When comparing the affected genes in the three conditions, it appears that 16 genes are common to all three (Fig 10), while 20 genes are common only to the 1g samples between days 5 and 6 (3g>1g and 3g>axe). Respectively, 69 and 20 genes are common between the static 1g for 1 day (3g>1g) or rotating 1g (3g>axe) for 1 day and the larvae having spent all 6 days at 1g (1g). Several genes, mostly common to all three conditions, were selected and the modulation of their expression was confirmed by RT-qPCR (S16 Table). Regulatory networks were constructed using the genes common to all three conditions, but also using those common to the 1g for one day condition (3g>1g and 3g>1axe) (Fig 11). Strikingly, a network composed of 7 genes (FOS, FOSB, EGR1, EDN1, SOCS3, GADD45B, KLF2) that were affected in exactly the same manner in all three conditions could be constructed, indicating that they represent a central network that is affected by gravitational conditions. Most importantly, these central genes were affected to the same extent, relative to the 3g for 6 days control, whether the larvae were kept at 1g during the entire experiment or only for the last day, suggesting that their expression levels are specific to this gravitational condition and are rapidly (within one day) adapted to new conditions. Five additional genes (MVP, HBE1, HES5, SOX10, LGALS3BP) were only affected after 1 day at lower gravity (both 3g>1g and 3g>1axe), indicating that they may be actually involved in the mechanism for rapid adaptation to lower gravity. Further analyses were performed using all the genes common to any two of the conditions (S5 Fig), also analyzed according to their potential function in individual organ systems (S6 Fig). By extending the network that way, other nodes become apparent, such as the nuclear receptor PPARG, the protein chaperone HSP90AA1 and the regulatory peptide endothelin (EDN1) (S5 Fig). Expression of NFKBIA, a target gene for the NFkB pathway coding for an inhibitor of this pathway, was decreased in two conditions, potentially causing the decreased expression of the antiproliferative factor BTG2 [80] observed in all three conditions.


Zebrafish Bone and General Physiology Are Differently Affected by Hormones or Changes in Gravity.

Aceto J, Nourizadeh-Lillabadi R, Marée R, Dardenne N, Jeanray N, Wehenkel L, Aleström P, van Loon JJ, Muller M - PLoS ONE (2015)

Network of genes affected in "relative microgravity" experiments.A network was constructed using the genes common to all three experiments, or the genes common only to 3g>1g and 3g>axe. Color overlay indicates the fold change relative to the 3g sample taken as control. Genes up-regulated (red), down-regulated (green), (*) indicates that the gene is represented by two or more probes on the microarray.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4465622&req=5

pone.0126928.g011: Network of genes affected in "relative microgravity" experiments.A network was constructed using the genes common to all three experiments, or the genes common only to 3g>1g and 3g>axe. Color overlay indicates the fold change relative to the 3g sample taken as control. Genes up-regulated (red), down-regulated (green), (*) indicates that the gene is represented by two or more probes on the microarray.
Mentions: When comparing the affected genes in the three conditions, it appears that 16 genes are common to all three (Fig 10), while 20 genes are common only to the 1g samples between days 5 and 6 (3g>1g and 3g>axe). Respectively, 69 and 20 genes are common between the static 1g for 1 day (3g>1g) or rotating 1g (3g>axe) for 1 day and the larvae having spent all 6 days at 1g (1g). Several genes, mostly common to all three conditions, were selected and the modulation of their expression was confirmed by RT-qPCR (S16 Table). Regulatory networks were constructed using the genes common to all three conditions, but also using those common to the 1g for one day condition (3g>1g and 3g>1axe) (Fig 11). Strikingly, a network composed of 7 genes (FOS, FOSB, EGR1, EDN1, SOCS3, GADD45B, KLF2) that were affected in exactly the same manner in all three conditions could be constructed, indicating that they represent a central network that is affected by gravitational conditions. Most importantly, these central genes were affected to the same extent, relative to the 3g for 6 days control, whether the larvae were kept at 1g during the entire experiment or only for the last day, suggesting that their expression levels are specific to this gravitational condition and are rapidly (within one day) adapted to new conditions. Five additional genes (MVP, HBE1, HES5, SOX10, LGALS3BP) were only affected after 1 day at lower gravity (both 3g>1g and 3g>1axe), indicating that they may be actually involved in the mechanism for rapid adaptation to lower gravity. Further analyses were performed using all the genes common to any two of the conditions (S5 Fig), also analyzed according to their potential function in individual organ systems (S6 Fig). By extending the network that way, other nodes become apparent, such as the nuclear receptor PPARG, the protein chaperone HSP90AA1 and the regulatory peptide endothelin (EDN1) (S5 Fig). Expression of NFKBIA, a target gene for the NFkB pathway coding for an inhibitor of this pathway, was decreased in two conditions, potentially causing the decreased expression of the antiproliferative factor BTG2 [80] observed in all three conditions.

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Laboratory for Organogenesis and Regeneration, GIGA- Research, University of Liège, B-4000, Liège, Sart-Tilman, Belgium.

ABSTRACT
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.

No MeSH data available.


Related in: MedlinePlus