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SUMMARYThe external cell layer that surrounds the fish primary myotome provides the myogenic precursors necessary for muscle growth, suggesting that this epithelium is equivalent to the amniote dermomyotome. In this study we report the identification of a trout orthologue of the dermal marker Dermo-1, and show that trout somitic external cells, which are all potentially myogenic as indicated by the transcription of Pax7 gene,express Dermo-1. This finding and our previous observation that external cells express collagen I show that these cells have dermis-related characteristics in addition to exhibiting myogenic features. In an effort to identify novel genes expressed in the external cell epithelium we performed an in situ hybridisation screen and found both collectin sub-family member 12,a transmembrane C-type lectin, and Seraf, an EGF-like repeat autocrine factor. In situ hybridisation of staged trout embryos revealed that the expression of Dermo-1, collectin sub-family member 12 and Seraf within the external cell layer epithelium was preceded by a complex temporal and spatial expression pattern in the early somite.

SUMMARY Myostatin (GDF8) has been shown to be a major genetic determinant of skeletal muscle growth in mammals. In this study, we report the cloning of two trout cDNAs that encode two distinct myostatin-related proteins. The presence in this fish species of two myostatin genes (Tmyostatin 1 and Tmyostatin 2) probably results from the recent tetraploïdisation of the salmonid genome. A comparative reverse-transcriptase-linked polymerase chain reaction assay revealed that Tmyostatin 1 mRNA was present ubiquitously in trout tissues, while Tmyostatin 2 mRNA expression was restricted to muscle and brain. In developing muscle, Tmyostatin 1 expression was observed in eyed-stage embryos well before hatching, whereas Tmyostatin 2 was expressed only in free-swimming larvae. In myotomal muscle from adult animals, Tmyostatin 1 mRNA accumulation was similar in both slow- and fast-twitch fibres, and its concentration did not change during the muscle wasting associated with sexual maturation. In contrast, Tmyostatin 2 mRNA accumulated predominantly in slow-twitch fibres, and its concentration decreased dramatically in wasting muscles from maturing animals. This work shows that two distinct myostatin genes are present in the trout genome. Furthermore, it indicates that these two trout myostatin genes (i) exhibit a distinct expression pattern in muscle and non-muscle tissues and (ii) are not upregulated during the muscle wasting that accompanies sexual maturation.

SUMMARYThe axial muscle of most teleost species consists of a deep bulk of fast-contracting white fibres and a superficial strip of slow-contracting red fibres. To investigate the embryological development of fast and slow muscle in trout embryos, we carried out single and double in situ hybridisation with fast and slow myosin heavy chain (MyHC)-isoform-specific riboprobes. This showed that the slow-MyHC-positive cells originate in a region of the somite close to the notochord. As the somite matures in a rostrocaudal progression, the slow-MyHC-positive cells appear to migrate radially away from the notochord to the lateral surface of the myotome, where they form the superficial strip of slow muscle. Surprisingly, the expression pattern of the fast MyHC showed that the differentiation of fast muscle commences in the medial domain of the somite before the differentiation and migration of the slow muscle precursors. Later, as the differentiation of fast muscle progressively spreads from the inside to the outside of the myotome, slow-MyHC-expressing cells become visible medially. Our observations that the initial differentiation of fast muscle takes place in proximity to axial structures and occurs before the differentiation and migration of slow muscle progenitors are not in accord with the pattern of muscle formation in teleosts previously described in the zebrafish Danio rerio, which is often used as the model organism in fishes.