Muscle fibre number varies with haemoglobin phenotype in Atlantic cod as predicted by the optimal fibre number hypothesis.

Atlantic cod (Gadus morhua L.) with the HbI-(2/2) haemoglobin phenotype have a higher blood oxygen affinity at low temperatures and a lower routine metabolic rate than individuals with the HbI-(1/1) phenotype. In the present study, muscle structure was found to be related to haemoglobin phenotype in a coastal population of Atlantic cod from the Saltenfjord region of Northern Norway. The maximum number of fast muscle fibres (FNmax) was reached at approximately 39 cm fork length and was 15% greater in the HbI-(1/1) than in the HbI-(2/2) phenotypes whereas the average fibre diameter for fish of the same fork length was significantly lower. Theoretically, the higher oxygen affinity of the HbI-(2/2) phenotype in the cold water of northern latitudes could have resulted in a relaxation of diffusional constraints at the level of individual muscle fibres, permitting the observed increase in fibre diameter. The results support the optimal fibre number hypothesis which envisages a trade-off between diffusional constraints and the energy cost of maintaining ionic homeostasis with fewer larger diameter muscle fibres in the HbI-(2/2) phenotype contributing to a lower routine metabolic rate.

Loss of muscle fibres in a landlocked dwarf Atlantic salmon population.

Growth of fast myotomal muscle in teleosts involves the continuous production of muscle fibres until some genetically pre-determined length. The dwarf landlocked (Bleke) population of Atlantic salmon (Salmo salar L.) from Byglands-fjord, Southern Norway mature at about 25 cm fork length and reach a maximum size of only 30 cm in the wild. The maximum diameter (D(max)) of fast muscle fibres in 4-year-old Bleke salmon (25-28 cm fork length) was 118 microm and not significantly different from that found in immature migratory salmon of a similar size. In contrast no evidence for active fibre recruitment was found in the Bleke salmon, such that the maximum fibre number, FN(max), was only 21-30% of that reported in typical farmed and wild migratory populations, respectively. We hypothesise that, once established, the physiological consequences of the dwarf condition led to rapid selection for reduced fibre number, possibly to reduce the maintenance costs associated with ionic homeostasis.

Rapid evolution of muscle fibre number in post-glacial populations of Arctic charr Salvelinus alpinus.

Thingvallavatn, the largest and one of the oldest lakes in Iceland, contains four morphs of Arctic charr Salvelinus alpinus. Dwarf benthic (DB), large benthic (LB), planktivorous (PL) and piscivorous (PI) morphs can be distinguished and differ markedly in head morphology, colouration and maximum fork length (FL(max)), reflecting their different resource specialisations within the lake. The four morphs in Thingvallavatn are thought to have been isolated for approximately 10 000 years, since shortly after the end of the last Ice Age. We tested the null hypothesis that the pattern of muscle fibre recruitment was the same in all morphs, reflecting their recent diversification. The cross-sectional areas of fast and slow muscle fibres were measured at 0.7 FL in 46 DB morphs, 23 LB morphs, 24 PL morphs and 22 PI morphs, and the ages of the charr were estimated using sacculus otoliths. In fish larger than 10 g, the maximum fibre diameter scaled with body mass (M(b))(0.18) for both fibre types in all morphs. The number of myonuclei per cm fibre length increased with fibre diameter, but was similar between morphs. On average, at 60 mum diameter, there were 2264 nuclei cm(-1) in slow fibres and 1126 nuclei cm(-1) in fast fibres. The absence of fibres of diameter 4-10 mum was used to determine the FL at which muscle fibre recruitment stopped. Slow fibre number increased with body length in all morphs, scaling with M(b)(0.45). In contrast, the recruitment of fast muscle fibres continued until a clearly identifiable FL, corresponding to 18-19 cm in the dwarf morph, 24-26 cm in the pelagic morph, 32-33 cm in the large benthic morph and 34-35 cm in the piscivorous morph. The maximum fast fibre number (FN(max)) in the dwarf morph (6.97x10(4)) was 56.5% of that found in the LB and PI morphs combined (1.23x10(5)) (P<0.001). Muscle fibre recruitment continued until a threshold body size and occurred at a range of ages, starting at 4+ years in the DB morph and 7+ years in the LB and PI morphs. Our null hypothesis was therefore rejected for fast muscle and it was concluded that the dwarf condition was associated with a reduction in fibre number. We then investigated whether variations in development temperature associated with different spawning sites and periods were responsible for the observed differences in muscle cellularity between morphs. Embryos from the DB, LB and PL morphs were incubated at temperature regimes simulating cold subterranean spring-fed sites (2.2-3.2 degrees C) and the general lakebed (4-7 degrees C). Myogenic progenitor cells (MPCs) were identified using specific antibodies to Paired box protein 7 (Pax 7), Forkhead box protein K1-alpha (FoxK1-alpha), MyoD and Myf-5. The progeny showed no evidence of developmental plasticity in the numbers of either MPCs or muscle fibres. Juveniles and adult stages of the DB and LB morphs coexist and have a similar diet. We therefore conclude that the reduction in FN(max) in the dwarf morph probably has a genetic basis and that gene networks regulating myotube production are under high selection pressure. To explain these findings we propose that there is an optimal fibre size, and hence number, which varies with maximum body size and reflects a trade-off between diffusional constraints on fibre diameter and the energy costs of maintaining ionic gradients. The predictions of the optimal fibre size hypothesis and its consequences for the adaptive evolution of muscle architecture in fishes are briefly discussed.

Reduction in muscle fibre number during the adaptive radiation of notothenioid fishes: a phylogenetic perspective.

The fish fauna of the continental shelf of the Southern Ocean is dominated by a single sub-order of Perciformes, the Notothenioidei, which have unusually large diameter skeletal muscle fibres. We tested the hypothesis that in fast myotomal muscle a high maximum fibre diameter (FD(max)) was related to a reduction in the number of muscle fibres present at the end of the recruitment phase of growth. We also hypothesized that the maximum fibre number (FN(max)) would be negatively related to body size, and that both body size and size-corrected FN(max) would show phylogenetic signal (tendency for related species to resemble each other). Finally, we estimated ancestral values for body size and FN(max). A molecular phylogeny was constructed using 12S mitochondrial rRNA sequences. A total of 16 species were studied from the Beagle Channel, Tierra del Fuego (5-11 degrees C), Shag Rocks, South Georgia (0.5-4 degrees C), and Adelaide Island, Antarctic Peninsula (-1.5 to 0.5 degrees C). The absence of muscle fibres of less than 10 micro m diameter was used as the criterion for the cessation of fibre recruitment. FD(max) increased linearly with standard length (SL), reaching 500-650 micro m in most species. Maximum body size was a highly significant predictor of species variation in FN(max), and both body size and size-corrected FN(max) showed highly significant phylogenetic signal (P<0.001). Estimates of trait values at nodes of the maximum likelihood phylogenetic tree were consistent with a progressive reduction in fibre number during part of the notothenioid radiation, perhaps serving to reduce basal energy requirements to compensate for the additional energetic costs of antifreeze production. For example, FN(max) in Chaenocephalus aceratus (12 700+/-300, mean +/- S.E.M., N=18) was only 7.7% of the value found in Eleginops maclovinus (164 000+/-4100, N=17), which reaches a similar maximum length (85 cm). Postembryonic muscle fibre recruitment in teleost fish normally involves stratified followed by mosaic hyperplasia. No evidence for this final phase of growth was found in two of the most derived families (Channichthyidae and Harpagiferidae). The divergence of the notothenioids in Antarctica after the formation of the Antarctic Polar Front and more recent dispersal north would explain the high maximum diameter and low fibre number in the derived sub-Antarctic notothenioids. These characteristics of notothenioids may well restrict their upper thermal tolerance, particularly for Champsocephalus esox and similar Channichthyids that lack respiratory pigments.