Bone without minerals and its secondary mineralization in Atlantic salmon (Salmo salar): the recovery from phosphorus deficiency.

Calcium and phosphorus (P) are the main bone minerals, and P deficiency can cause hypomineralized bones (osteomalacia) and malformations. This study used a P-deficient salmon model to falsify three hypotheses. First, an extended period of dietary P deficiency does not cause pathologies other than osteomalacia. Second, secondary mineralization of non-mineralized bone is possible. Third, secondary mineralization can restore the bones' mineral composition and mechanical properties. For 7 weeks, post-smolt Atlantic salmon (Salmo salar) received diets with regular P content (RP) or with a 50% lowered P content (LP). For additional 9 weeks, RP animals continued on the regular diet (RP-RP). LP animals continued on the LP diet (LP-LP), on a regular P diet (LP-RP) or on a high P diet (LP-HP). After 16 weeks, animals in all groups maintained a non-deformed vertebral column. LP-LP animals continued bone formation albeit without mineralization. Nine weeks of RP diet largely restored the mineral content and mechanical properties of vertebral bodies. Mineralization resumed deep inside the bone and away from osteoblasts. The history of P deficiency was traceable in LP-RP and LP-HP animals as a ring of low-mineralized bone in the vertebral body endplates, but no tissue alterations occurred that foreshadow vertebral body compression or fusion. Large quantities of non-mineralized salmon bone have the capacity to re-mineralize. If 16 weeks of P deficiency as a single factor is not causal for typical vertebral body malformations, other factors remain to be identified. This example of functional bone without minerals may explain why some teleost species can afford to have an extremely low mineralized skeleton.

Assessment of global proteome dynamics in carp: a model for investigating environmental stress.

Fish have to respond to a range of natural and man-made environmental stressors, which can lead to molecular changes within their tissues. Many studies focused on environmental stress in fish have examined the change in protein abundance or mRNA level. However, it is well-known that there is a disconnect between mRNA and protein expression. In order to bridge this gap, protein turnover must also be considered. We have developed an experimental strategy to determine the synthesis rates of individual proteins in the tissues of fish on a proteome-wide scale. This approach has been applied to the common carp ( Cyprinus carpio ), a key model species for investigating environmentally induced physiological plasticity. We have calculated the rates of protein synthesis for over a thousand individual proteins from the skeletal muscle and liver of carp. The median synthesis rate of proteins from liver was higher than that of skeletal muscle. The analysis further revealed that the same protein can have a different rate of synthesis depending on the tissue type. Our strategy permits a full investigation of proteome dynamics in fish and will have relevance to the fields of integrative biology and ecotoxicology.

Temperature sensitive regions of the Chinook salmon vertebral column: Vestiges and meristic variation.

Variation of vertebral centra numbers is common in vertebrates. Likewise, the number of associated elements such as ribs and neural and haemal arches can vary and affect all regions of the vertebral column. In mammals, only the number of cervical vertebrae is invariable. Variation of total vertebral centra numbers is well documented in teleost fish, often related to temperature. Less information is available about which part of the vertebral column and which associated elements are liable to variation. Here, variation in number of vertebral centra and associated elements is studied in Chinook salmon in six distinct anatomical regions. Animals are raised at 8 and 12°C to ask whether the vertebral centrum numbers, the pattern, and the frequency of variation in particular regions are temperature dependent. No significant difference concerning the total number of vertebrae was found, but regional differences occurred between the 8 and 12°C groups. Twelve specimens out of 60 of the 12°C group had three postcranial vertebrae compared to only one specimen in the 8°C group. The number of transitional vertebrae is significantly different in 8 and 12°C specimens. Fewer transitional vertebrae occur in more anterior positions in 8°C specimens. Most specimens of both temperature groups had two ural centra; however 17 specimens out of 60 of the 12°C group had up to five ural centra. Specimens of the 12°C group show more variation in the presence of the vestigial ribs associated with transitional vertebrae. Clearly, the postcranial, transitional, and ural regions are temperature sensitive. This study shows that nonsignificant differences in the total number of vertebrae can mask significant regional variation. Variation of vertebral numbers could be the consequence of loss or gain of vertebral centra and/or a change in the identity of the associated element on the vertebral centrum.

A proteomics strategy for determining the synthesis and degradation rates of individual proteins in fish.

In order to study the protein dynamics in the tissues of fish we have developed a proteomics-based strategy to determine the rates of synthesis and degradation of individual proteins. We have demonstrated the feasibility of this approach by measuring the turnover of multiple isoforms of parvalbumin (ß1-7) in the skeletal muscle of common carp (Cyprinus carpio). A stable isotope-labelled amino acid ([(2)H(7)] l-leucine) was administered to the carp via the diet and its incorporation into the isoforms of parvalbumin in muscle over time was monitored by LC-MS analysis of signature peptides. The relative isotope abundance was calculated and used to deconvolute the data. The ß7 parvalbumin isoform had a rate of synthesis that was greater than the rate of degradation. In contrast the rate of degradation of the ß5 isoform exceeded its rate of synthesis, whilst the analysis revealed that the other parvalbumin ß-isoforms (ß1, ß2, ß3, ß4 and ß6) had a rate of synthesis that was equal to the rate of degradation. This work has addressed a number of technical challenges and represents the first study to use proteomic approaches to measure the turnover of individual proteins in fish.