Nuclear DNA content variation associated with muscle fiber hypertrophic growth in decapod crustaceans

Abstract

We tested the hypothesis that hypertrophic muscle growth in decapod crustaceans is associated with increases in both the number of nuclei per fiber and nuclear DNA content. The DNA-localizing fluorochrome DAPI (4',6-diamidino-2-phenylindole) and chicken erythrocyte standards were used with static microspectrophotometry and image analysis to estimate nuclear DNA content in hemocytes and muscle fibers from eight decapod crustacean species: Farfantepenaeus aztecus, Palaemonetes pugio, Panulirus argus, Homarus americanus, Procambarus clarkii, Cambarus bartonii, Callinectes sapidus, and Menippe mercenaria. Mean diploid (2C) values in hemocytes ranged from 3.6 to 11.7 pg. Hemocyte 2C estimates were used to extrapolate ploidy level in the multinucleated skeletal muscle tissue of juvenile and adult animals. Across all species, mean muscle fiber diameters from adult animals were significantly larger than those in juveniles, and nuclear domains were greater in larger fibers. The number of nuclei per fiber increased with increasing fiber size, as hypothesized. Maximum nuclear DNA content per species in muscle ranged from 4C to 32C, consistent with endopolyploidy. Two patterns of body- and fiber-size-dependent shifts in ploidy were observed: four species had a significantly higher ploidy in the larger fibers of adults, while three species exhibited a significantly lower ploidy in adults than in juveniles. Thus, across species, there was no systematic relationship between nuclear domain size and nuclear DNA content.