1H NMR metabolic profiling of cod (Gadus morhua) larvae: potential effects of temperature and diet composition during early developmental stages.

Marine aquaculture offers a great source of protein for the increasing human population, and farming of, for example, Atlantic salmon is a global industry. Atlantic cod farming however, is an example of a promising industry where the potential is not yet realized. Research has revealed that a major bottleneck to successful farming of cod is poor quality of the larvae and juveniles. A large research program was designed to increase our understanding of how environmental factors such as temperature and nutrition affects cod larvae development. Data on larvae growth and development were used together with nuclear magnetic resonance. The NMR data indicated that the temperature influenced the metabolome of the larvae; differences were related to osmolytes such as betaine/TMAO, the amino acid taurine, and creatine and lactate which reflect muscle activity. The larvae were fed Artemia from stage 2, and this was probably reflected in a high taurine content of older larvae. Larvae fed with copepods in the nutrition experiment also displayed a high taurine content, together with higher creatine and betaine/TMAO content. Data on the cod larvae metabolome should be coupled to data on gene expression, in order to identify events which are regulated on the genetic level versus regulation resulting from temperature or nutrition during development, to fully understand how the environment affects larval development.

Differential spatio-temporal expression and functional diversification of the myogenic regulatory factors MyoD1 and MyoD2 in Atlantic halibut (Hippoglossus hippoglossus).

Development of the vertebrate skeletal muscle is orchestrated by the myogenic regulatory factors MyoD, Myf5, myogenin and MRF4, which likely arose from the duplications of a single ancestral gene early in vertebrate evolution. We have isolated two myod genes from Atlantic halibut and examined their differential expression during embryogenesis using quantitative PCR and in situ hybridization to address their functional roles in this asymmetrically organized flatfish. myod1 was initially maternally expressed, while myod2 mRNA was first detectable during gastrulation. The myod1 mRNA levels predominated throughout somitogenesis, and both slow and fast muscle precursor cells displayed the bilateral symmetric myod1 signal during the formation of the symmetric somite pairs. In contrast, myod2 was left-right asymmetrically expressed in the fast muscle precursors. The random expression of myod2 was not associated with the right-sided eye migration and the development of thicker fast skeletal muscle on the eyed side than on the blind side. The nucleotide substitution analysis indicated that the teleost MyoDs essentially have evolved under purifying selection, but a subset of amino acid sites under strong positive selection were identified in the MyoD2 branch. Altogether, halibut MyoD1 seems to have retained the central role of MyoD in driving skeletal myogenesis, whereas the function of MyoD2 is uncertain in this flatfish species.

Molecular characterization of a PDZ-LIM protein in Atlantic salmon (Salmo salar): a fish ortholog of the alpha-actinin-associated LIM-protein (ALP).

A protein containing both PDZ and LIM protein-protein interaction motifs has for the first time been identified in a lower vertebrate species. A full-length cDNA encoding the ortholog of the alpha-actinin-associated LIM protein (ALP) was isolated from white skeletal muscle of Atlantic salmon (Salmo salar). Whereas ALP is expressed as two muscle specific isoforms in mammals and chicken as the result of alternative splicing, a single ALP transcript was found in both muscle and non-muscular tissues of Atlantic salmon. On the other hand, Western blot analysis revealed several immunoreactive ALP variants in salmon muscle tissues, including a 45 kDa protein in white and red skeletal muscle and a 37-40 kDa protein in heart and smooth muscle. Salmon ALP and alpha-actinin showed similar striated patterns in serial longitudinal sections of white and red skeletal muscle and heart muscle. Expression of ALP was initiated at the 45-somite stage of the salmon embryogenesis contemporary with the first appearance of alpha-actinin transcripts. The similarities in both the spatial and temporal expression patterns of salmon ALP and alpha-actinin strongly indicate that the two proteins are associated as in higher vertebrates, and that the assumed involvement of ALP in the organization and/or maintenance of the Z-lines in striated muscle has been conserved during vertebrate evolution. However, in contrast to the restricted expression of ALP in higher vertebrates, the ubiquitous expression of salmon ALP suggest that this factor is involved in the assembly of additional multi-protein complexes in fish.