Morphological and molecular characterization of developing vertebral fusions using a teleost model

<p>Abstract</p> <p>Background</p> <p>Spinal disorders are a major cause of disability for humans and an important health problem for intensively farmed animals. Experiments have shown that vertebral deformities present a complex but comparable etiology across species. However, the underlying molecular mechanisms involved in bone deformities are still far from understood. To further explicate the mechanisms involved, we have examined the fundamental aspects of bone metabolism and pathogenesis of vertebral fusions in Atlantic salmon (<it>Salmo salar</it>).</p> <p>Results</p> <p>Experimentally, juvenile salmon were subjected to hyperthermic conditions where more than 28% developed fused vertebral bodies. To characterize the fusion process we analyzed an intermediate and a terminal stage of the pathology by using x-ray, histology, immunohistochemistry, real-time quantitative PCR and <it>in situ </it>hybridization. At early stage in the fusion process, disorganized and proliferating osteoblasts were prominent at the growth zones of the vertebral body endplates. PCNA positive cells further extended along the rims of fusing vertebral bodies. During the developing pathology, the marked border between the osteoblast growth zones and the chondrocytic areas connected to the arches became less distinct, as proliferating cells and chondrocytes blended through an intermediate zone. This cell proliferation appeared to be closely linked to fusion of opposing arch centra. During the fusion process a metaplastic shift appeared in the arch centra where cells in the intermediate zone between osteoblasts and chondrocytes co-expressed mixed signals of chondrogenic and osteogenic markers. A similar shift also occurred in the notochord where proliferating chordoblasts changed transcription profile from chondrogenic to also include osteogenic marker genes. In progressed fusions, arch centra and intervertebral space mineralized.</p> <p>Conclusion</p> <p>Loss of cell integrity through cell proliferation and metaplastic shifts seem to be key events in the fusion process. The fusion process involves molecular regulation and cellular changes similar to those found in mammalian deformities, indicating that salmon is suitable for studying general bone development and to be a comparative model for spinal deformities.</p

Molecular pathology of vertebral deformities in hyperthermic Atlantic salmon (Salmo salar)

<p>Abstract</p> <p>Background</p> <p>Hyperthermia has been shown in a number of organisms to induce developmental defects as a result of changes in cell proliferation, differentiation and gene expression. In spite of this, salmon aquaculture commonly uses high water temperature to speed up developmental rate in intensive production systems, resulting in an increased frequency of skeletal deformities. In order to study the molecular pathology of vertebral deformities, Atlantic salmon was subjected to hyperthermic conditions from fertilization until after the juvenile stage.</p> <p>Results</p> <p>Fish exposed to the high temperature regime showed a markedly higher growth rate and a significant higher percentage of deformities in the spinal column than fish reared at low temperatures. By analyzing phenotypically normal spinal columns from the two temperature regimes, we found that the increased risk of developing vertebral deformities was linked to an altered gene transcription. In particular, down-regulation of extracellular matrix (ECM) genes such as <it>col1a1</it>, <it>osteocalcin</it>, <it>osteonectin </it>and <it>decorin</it>, indicated that maturation and mineralization of osteoblasts were restrained. Moreover, histological staining and <it>in situ </it>hybridization visualized areas with distorted chondrocytes and an increased population of hypertrophic cells. These findings were further confirmed by an up-regulation of <it>mef2c </it>and <it>col10a</it>, genes involved in chondrocyte hypertrophy.</p> <p>Conclusion</p> <p>The presented data strongly indicates that temperature induced fast growth is severely affecting gene transcription in osteoblasts and chondrocytes; hence change in the vertebral tissue structure and composition. A disrupted bone and cartilage production was detected, which most likely is involved in the higher rate of deformities developed in the high intensive group. Our results are of basic interest for bone metabolism and contribute to the understanding of the mechanisms involved in development of temperature induced vertebral pathology. The findings may further conduce to future molecular tools for assessing fish welfare in practical farming.</p

The effects of carbon dioxide on growth performance, welfare, and health of Atlantic salmon post-smolt (Salmo salar) in recirculating aquaculture systems

High carbon dioxide (CO2) concentrations negatively impact fish, which makes data on its tolerance especially relevant for production systems that can accumulate CO2 such as recirculating aquaculture system (RAS). The current study evaluates the effect of CO2 on the growth performance, welfare, and health of Atlantic salmon post-smolts in RAS. This study consisted of two phases. The first was a CO2 exposure phase, where eighteen tanks were used with six treatments in triplicate: 5, 12, 19, 26, 33 and 40 mg/L of CO2 during 12 weeks in a 12 ppt salinity RAS (hereafter RAS phase). In the second phase, PIT-tagged fish were transferred to a 34 ppt salinity single flow-through tank at CO2 2 was low and not related to treatments. The mean final body weight was significantly higher in the 5 mg/L treatment compared to CO2 treatments ≥12 mg/L at the end of RAS phase and to CO2 treatments ≥33 mg/L at the end of seawater phase. Moreover, regressions showed that growth significantly decreased linearly with increasing CO2 in the water. Eye cataracts and visible external damage on skin, operculum, and fins were inexistent and similar among CO2 treatments. Kidneys showed no signs of mineral deposits in any of the structures of the tissue. However, skin analysis showed that fish exposed to high CO2 concentrations had a significantly thinner dermis layer (both at the end of RAS and seawater phase) and a significantly thinner epidermis layer and lower mucus cells count (at the end of seawater phase). In conclusion, Atlantic salmon post-smolts cultured in brackish water RAS showed a maximum growth performance at CO2 concentrations below 12 mg/L. Except skin, no major effects of health and welfare were observed, including cataracts and nephrocalcinosis. Further studies should evaluate the molecular and physiological responses to both short-term and long-term carbon dioxide exposure.</p

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