Muscle cellularity was studied in Pagellus bogaraveo juveniles fed on diets with different protein contents. Measured in transversal body sections, at both post-opercular and post-anal locations, the morphometric variables estimated were: total muscle area (A), total number of fibres (N), number of fibres per unit area of muscle (N A ) and cross-sectional fibre area ð" aÞ; of the two main muscle fibre types. At the end of the experiment fish fed on diets having more than 40% of protein displayed significantly higher body weight. Fish fed on protein-rich diets exhibited greater " a and N. For fish fed on 30 and 50% protein diets the morphometric parameters measured grew linearly with the fish weight. High-protein diets favoured muscle hyperplasia. When comparing rostral and caudal locations, a greater N and a smaller " a of posterior red fibres were the consistent differences found-a fact, to our knowledge, so far unreported for fish.

Sex differences exist in fish hepatocytes, but studies for characterizing their cytology throughout the breeding cycle are still scarce; suggesting changes, but most lacking quantitative data. To address this limitation, to complement baseline data generated from the brown trout model, and to prove that sex-specific seasonal changes exist, we made an unbiased stereological evaluation of the hepatocytic cytoplasm. Unprecedentedly for fish liver, the stereological design was exempt from model (biased) assumptions. Five (3 years old) animals per sex were studied in endogenous vitellogenesis, exogenous vitellogenesis, and spawning season end. Liver pieces for analysis were systematically sampled. Stereology was done in transmission electron microscopy (TEM) micrographs. Primary data generated relative volume estimates of the major cytoplasmic components. Such values were used for deriving absolute volumes (per cell and per liver). Lipid droplets did not show changes. As to other targets, trends at cell and liver levels were not always equal. If the hepatocyte was the reference space, the contents in mitochondria, dense bodies, glycogen, and cytosol changed seasonally, in both sexes. If taking the liver as the reference, changes attained the Golgi apparatus and rough endoplasmic reticulum (RER), besides dense bodies, glycogen (in females), and cytosol. The components volumes (namely per liver) were often positively (negatively for glycogen) correlated with the ovary weight, disclosing new associations and implications in fish. While also offering gold-standard data for backing morphofunctional correlations and pathology, we revealed a new process by which females increase the amount of RER and Golgi throughout vitellogenesis, breaking from the idea on how this event happens in fish.

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To understand better the growth mechanisms in the economically important fish Pagellus bogaraveo, in terms of muscle fibre hyperplasia v. hypertrophy, the lateral muscle of this fish was studied morphometrically from hatching to juvenile comparing rostral and caudal locations. Fish were sampled at 0, 5, 23, 40, 70, 100, 140 and 180 days. Fibre types were first identified by succinate dehydrogenase (SDH) and immunostaining with a polyclonal antibody against fish slow myosin (4-96). Morphometric variables were then measured in transverse body sections, at both post-opercular and post-anal locations, to estimate the following variables: total muscle area [A (muscle)], total fibre number [N (fibres)], fibre number per unit area of muscle [N(A)(fibres, muscle)] and cross-sectional fibre area [a (fibres)] of the two main muscle fibre types (white and red). Overall, growth throughout the various stages resulted from increases both in the number and in the size of muscle fibres, paralleled by an expansion of the [A (muscle)]. Nonetheless, that increase was not significant between 0-5 days on one hand and 100-140 days, on the other hand. On the contrary, the [N(A)(fibres, muscle)] declined as the body length increased. Analysis of the muscle growth kinetics suggested that, within the important time frame studied, hyperplasia gave the main relative contribution to the increase of white muscle [A (white muscle)], whereas red muscle [A (red muscle)] mainly grew by hypertrophy, with both phenomena occurring at a faster pace posteriorly in the body. Finally, when comparing rostral and caudal locations, a greater [N (fibres)] and [A (muscle)] of the posterior white and red fibres were the consistent features. It was also observed that the proportion of the cross-sectional area of the myotomal muscle comprised of white muscle was greater in the anterior part of the fish.

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