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First published online June 15, 2006
Journal of Experimental Biology 209, 2432-2441 (2006)
Published by The Company of Biologists 2006
doi: 10.1242/jeb.02269

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Somite formation and expression of MyoD, myogenin and myosin in Atlantic halibut (Hippoglossus hippoglossus L.) embryos incubated at different temperatures: transient asymmetric expression of MyoD

Trina F. Galloway^1,*, Tora Bardal¹, Sylvia N. Kvam¹, Stine W. Dahle¹, Gaute Nesse², Markus Randøl², Elin Kjørsvik¹ and Øivind Andersen²

¹ Department of Biology, Brattøra Research Centre, Norwegian University of Science and Technology (NTNU), N-7491 Trondheim, Norway
² Institute of Aquaculture Research (AKVAFORSK), PO Box 5010, N-1430 Ås, Norway

View larger version (30K): [in a new window]   Fig. 1. Atlantic halibut MyoD cDNA and predicted amino acid sequences, which are numbered on the right. The bHLH domain is shaded. The locations of the two introns in the genomic sequence are indicated.

View larger version (28K): [in a new window]   Fig. 2. Atlantic halibut myogenin cDNA and predicted amino acid sequences, which are numbered on the right. The bHLH domain is shaded. The locations of the two introns in the genomic sequence are indicated. Alternative splicing by skipping exon 2 (underlined) generated a shorter transcript because of a premature stop codon (bold) in the changed reading frame.

View larger version (46K): [in a new window]   Fig. 3. Alignment of the conserved Cys-rich motif and the boxed bHLH and HIII of Atlantic halibut MyoD and myogenin and their orthologues from gilthead seabream (GenBank accession no. AAL85338), zebrafish (AAK06755, AAM82616), Atlantic cod (AAK17969), pufferfish (AAV70941), common carp (BAA33565, BAA33564), rainbow trout (CAA87010), Xenopus (CAA34232, AAL05070) and mouse (NP_112466, NP_034996). Conserved residues of both MyoD and myogenin are shaded in dark grey; distinct residues of the two factors are shaded in light grey. The major determinant of the differential functions of MyoD and myogenin is indicated by an arrow (Bergstrom and Tapscott, 2001).

View larger version (18K): [in a new window]   Fig. 4. Somite formation at different egg incubation temperatures. Each point represents one embryo/larva. Sigmoid curves were fitted for each incubation temperature [y=a/(1+e–(x–x0/b)), where y refers to number of somites and x to number of d° after fertilisation].

View larger version (71K): [in a new window]   Fig. 5. Spatial and temporal expression of MyoD, myogenin and MyHC mRNA in halibut embryos and larvae with similar numbers of somites. All specimens are dorsal views with the head towards the upper right corner of the picture. Arrowheads indicate the first somite with expression in lateral cells; arrows, the first somite with expression in adaxial cells; dot, asymmetric expression. Scale bar, 0.5 mm for all images.

View larger version (43K): [in a new window]   Fig. 6. Temporal expression of MyLC2a and MyLC2b during somitogenesis in Atlantic halibut. The presence of reverse-transcribed cDNAs was verified by amplifying ß-actin cDNA.