Histochemistry and morphometric analysis of muscle fibers from patients with duchenne muscular dystrophy (DMD) / Histoquímica y análisis morfométrico de las fibras musculares de p0acientes con distrofia muscular de duchenne (DMD)

The aim of the study was to analyze the muscle fibers by histochemistry and morphometric methods from patients with Duchenne muscular dystrophy (DMD). Muscle biopsies were taken from the vastus lateralis muscle of five boys between 13 and 15-years of age, with clinical diagnosis of DMD. The histochemistry was performed using myofibrillar ATPases (9.6, 4.6 and 4.3). To morphometrical analysis a computerized semiautomatic system and software Image-Lab was used. ATPase staining showed atrophy of muscle fibers. Fibrosis and adipose deposition occurred in variable degree depending of muscular involvement. The morphometrical analysis showed an increase of size (percentage) to type I fiber than other types in all patients. Furthermore, the type I fiber had a larger cross-sectional area and mean diameter than type IIa and IIb fibers. Both histochemistry and morphometric analysis could be important tools for qualitative and quantitative diagnostics of muscle fibers attacked in this type of disease.

El objetivo del estudio fue analizar las fibras musculares mediante histoquímica y métodos morfométricos en pacientes con distrofia muscular de Duchenne (DMD). Se tomaron biopsias musculares del músculo vasto lateral de cinco niños entre 13 y 15 años de edad, con diagnóstico clínico de DMD. La histoquímica se realizó mediante ATPasa miofibrilar (9.6, 4.6 y 4.3). Para el análisis morfométrico se utilizó un sistema semiautomático computarizado y software de imagen de laboratorio. La tinción de ATPasa mostró una atrofia de las fibras musculares. La fibrosis y depósito adiposo se observó en grado variable dependiendo del compromiso muscular. El análisis morfométrico mostró un aumento de tamaño (porcentaje) de fibras tipo I en todos los pacientes. Además, la fibra tipo I tuvo un área de sección transversal y diámetro medio mayor que las fibras tipos IIa y IIb. Tanto la histoquímica y el análisis morfométrico pueden ser herramientas importantes para el diagnóstico cualitativo y cuantitativo de las fibras musculares comprometidas en este tipo de enfermedad.

Morfologia e morfometria dos tipos de fibras do músculo reto abdominal de cadelas (Canis familiaris) durante a gestação / Morphologic and morphometry of rectus abdominis muscle fiber types of the during the gestation dog (Canis familioris)

Neste trabalho foram avaliadas as alterações morfológicas e o grau de hipertrofia das fibras de contração lentas (tipo I) e rápidas (tipo II) do músculo reto abdominal de cadelas adultas nas fases: pré-gestacional, 30 e 60 dias de gestação e 30 dias pós-parto (pp). Em todas essas fases, sob anestesia, foram removidas biópsias musculares, as quais foram congeladas. Cortes histológicos foram obtidos em um micrótomo e corados com hematoxilina e eosina (HE) para a avaliação da morfologia. Outros cortes foram submetidos à ATPase miofibrilar (m-ATPase), após pré-incubação em meio alcalino (pH = 10,4) e em meio ácido (pH = 4,4), visando à identificação das fibras tipo I e II. Nas cadelas virgens, o tecido muscular revelou-se constituído por fibras com diferentes diâmetros, com contornos poligonais, tendo um ou mais núcleos periféricos. Estas características foram similares nos animais com 30 dias de gestação. Aos 60 dias, ao lado de fibras com aparência normal, observaram-se fibras com contornos mais polimórficos e outras fibras de diâmetros muito pequenos. Aos 30 dias pp, as características do tecido foram similares às dos animais em fase pré-gestacional. Em todas as fases estudadas, os diâmetros fibras tipo II foram maiores do que as do tipo I. Aos 30 dias observou-se a redução significativa dos diâmetros de ambos os tipos de fibras e aumento significativo aos 60 dias de gestação. Durante a gestação, o estímulo crônico, representado pela expansão da parede abdominal, promoveu alterações da morfologia e morfometria das fibras.

The present paper deals with the evaluation of morphological and morphometric alterations of slow-twist (I) and fast-twitch (II) fibers of rectus abdominis muscle of adult female dog during pre-gestational phase, at 30 and 60 days of pregnancy and at 30 days after the porturition. At the every phases, using the open biopsy technique, muscle samples were collected. The samples were plunged. Histological sections were cut in a microtome. For general morphology, some sections were stained with HE. Subsequent sections were reacted for myofibrillor ATPase (m-ATPase), after alkaline (pH 10.4) and acid (pH 4.4) pre-incubations, in order to identificate type I and II fibers. In the pre-gestational phase, muscle tissue revealed to be composed by fibers with different diameters, presenting polygonal outlines and one or more periphery nuclei. At 30 days of pregnancy, muscle fiber characteristics were similar. At 60 days, in addition to the existence of normal fibers, polymorphic and small diameter fibers were frequent. At 30 days after the parturation, the morphology of muscle fiber were similar to that observed in the pre-gestational phase. In the four phases, type II fiber diameters were lager than type I. The diameters of both fiber types showed a significant reduction in the 30 days phase and a significative increasing at 60 days. The expansion of the abdominal wall during the pregnancy represents a chronic stimulus, induced changes in the morphology and in the fiber type diameters.