Functional morphology and fiber types of the masseter muscles of two caviomorph rodents with contrasting lifestyles, Ctenomys talarum (Ctenomyidae) and Cavia aperea (Caviidae).

The aim of this work is the analysis of histochemical and morphometric properties of the masseter muscles of Ctenomys talarum and Cavia aperea. The former belongs to a subterranean rodent clade, Ctenomyidae, which has evolved a robust masticatory apparatus adapted to chisel-tooth digging and processing of abrasive grasses; C. aperea belongs to the family Caviidae, with relatively graceful jaws and mandibular musculatures, consistent with less mechanically challenging diets. Adult males were captured, immediately transported to the laboratory, and euthanized in a CO2 chamber. The musculus masseter superficialis and musculus masseter profundus on the left side of the animals were used to analyze the histochemical composition of the fiber types treated with myosin adenosine triphosphatase, succinate dehydrogenase and periodic acid Schiff. The mean fiber diameters, relative areas, and frequencies of each muscle fiber type were calculated. The mm. masseter superficialis and masseter profundus on the right side were used to measure the physiological cross-sectional area (PCSA). Based on this measurement, the internal force (F) was estimated. In the m. masseter profundus of both species and in the m. masseter superficialis of C. aperea intermediate fast oxidative-glycolytic fibers (FOGi) predominated. In the mm. masseter superficialis and masseter profundus of C. talarum the relative area of fast glycolytic (FG) fibers was greater than that of the muscles of C. aperea, whose main muscle fiber component is FOGi fibers. When corrected for body mass differences, PCSA was higher for the mm. masseter superficialis of C. talarum. This and the larger relative area of FG fibers, probably contributes to the exertion of large bite forces in C. talarum, as measured in previous studies.

Functional morphology and identity of the thenar pad in the subterranean genus Ctenomys (Rodentia, Caviomorpha).

As in many other fossorial tetrapods, the most obvious adaptations to scratch-digging in the subterranean tuco-tuco (Rodentia, Ctenomyidae, Ctenomys) are found in the hands, which among other adaptations, present the mesaxonic condition; i.e. the central digits are more developed, and also their claws, which are curved and elongated. The thumb is atrophied and aligned with the rest of the digits, showing a small and flat claw. This configuration of digits and claws seems to be in accordance with what it is expected for rodents: rudimentary movements when handling food items. However, on the palmar side of the hand, tuco-tucos have several pads, the thenar (located under the thumb) being the most developed. In this study, we investigated the functional morphology of the thenar pad through different approaches: musculoskeletal anatomy, histology and functionality. The analysis of radiographs and clarified and double-stained hand samples of Ctenomys talarum and C. australis showed that the thenar pad is supported by a paddle-shaped bone that articulates with a protrusion in the scapholunate bone. This bone, flat and long, continues in a flat cartilaginous structure, with a shape similar to a claw. Dissections showed that the thenar pad has several associated muscles: the m. palmaris longus, the m. abductor pollicis longus, and a massive muscular complex located between the thumb and the thenar pad. By topology it might be inferred that this complex is formed by the m. abductor pollicis brevis, the m. flexor pollicis brevis and the m. adductor pollicis brevis. Longitudinal histological sections of the thenar pad stained with hematoxylin-eosin showed a thick layer of keratin at the distal end, external face. The observation of live specimens of C. talarum foraging on two food items of different size and filmed at 300 fps showed that the thenar pad acts as an opposable thumb, with digit-like movements. Tuco-tucos are able to perform more precise movements than expected, and to grasp and manipulate the food with one hand. In previous studies, it was suggested that the thenar pad was supported by a 'palmar ossicle', or 'prepollex' (= radial sesamoid bone). Our results suggest that this sesamoid underwent a radical change on its morphology, making the thenar pad a part of the food handling system in Ctenomys, so the thenar pad might be considered a 'false thumb', rather than a palmar pad. It is suggested to advance on the description and functional analysis of the thenar pad, redefining the structure, since the terms used so far to define it would not be accurate.

Influence of reduced salinity on the histochemical fiber type composition of the claw closer muscle of two estuarine crab species, Cyrtograpsus angulatus and Neohelice granulata (Decapoda: Varunidae) / Influencia de la salinidad reducida sobre la composición histoquímica de tipos de fibras del músculo de cierre de las quelas de dos especies de cangrejos estuariales, Cyrtograpsus angulatus y Neohelice granulata (Decapoda: Varunidae)

The estuarine crustaceans are exposed to frequent and abrupt environmental salinity changes that involve adjustments at different levels in muscle tissue. The histochemical composition of fiber types of the claw closer muscle of Cyrtograpsus angulatus and Neohelice granulata (Dana, 1851) under hyper regulatory conditions was analyzed. Cryosections of the muscle were treated with myosin-adenosine triphosphatase (m-ATPase), succinic dehydrogenase (SDH), periodic acid Schiff (PAS) and Sudan Black B. The mean diameters, the relative area and the proportion of each muscle fiber type were calculated. Types I and IV would belong to 'extreme' groups, whereas types II and III would be considered 'intermediate'. Type I fibers were large and exhibited a weak reaction to all techniques; type IV fibers were small and reacted strongly to histochemical tests. Types II and III prevailed in C. angulatus whereas type II predominated in N. granulata. Type IV fibers were absent in C. angulatus and scarce in N. granulata. The claw closer muscle of N. granulata and C. angulatus exhibited differential responses under reduced salinity at a histochemical level. Therefore, the existence of different adjustment mechanisms facing salinity stress is suggested.

Los crustáceos estuariales están expuestos a cambios frecuentes y abruptos en la salinidad ambiental que requieren ajustes a diferentes niveles en el tejido muscular. Se analizó la composición histoquímica de tipos de fibras del músculo de cierre de las quelas de Cyrtograpsus angulatus y Neohelice granulata (Dana, 1851) en condiciones de hiperregulación. Secciones de crióstato fueron tratadas con miosín-adenosín trifosfatasa (m-ATPasa), succinato deshidrogenasa (SDH), ácido periódico Schiff (PAS) y Sudan Black B. Se calcularon el diámetro promedio, el área y la proporción relativa de cada tipo de fibra. Los tipos I y IV serían grupos "extremos", mientras que los tipos II y III "intermedios". Las fibras del tipo I, de gran tamaño, exhibieron una reacción débil con todas las técnicas; las fibras del tipo IV, pequeñas, reaccionaron intensamente frente a los tests histoquímicos. Los tipos II y III predominaron en C. angulatus, mientras que el tipo II fue el más abundante en N. granulata. Las fibras del tipo IV fueron escasas en N. granulata y no se observaron en C. angulatus. El músculo de cierre de las quelas de N. granulata y C. angulatus exhibió respuestas diferenciales a nivel histoquímico en condiciones de salinidad reducida. De este modo, se sugiere la existencia de diferentes mecanismos de ajuste frente al estrés salino.

Histological and Histochemical Study of the Hepatopancreas of Two Estuarine Crab Species, Cyrtograpsus angulatus and Neohelice granulata (Grapsoidea, Varunidae): Influence of Environmental Salinity.

The aim of this study was to analyze the histology and the histochemical distribution of glycoproteins (GPs) and lipids of the hepatopancreas of Cyrtograpsus angulatus and Neohelice granulata acclimated to salinities of 10 psu (hyperregulation) and 35 psu (osmoconformation). Sections of the hepatopancreas of adult male crabs were treated with hematoxylin-eosin; Masson trichrome; Alcian Blue (pHs 2.8, 1.0, 0.5); Toluidine Blue (pHs 5.6, 4.2); periodic acid Schiff; Sudan Black and Red. At salinity 35 psu, the hepatopancreas of both species exhibited typical histological features, whereas at salinity 10 psu, detachment of the basal lamina, desquamated epithelium, disrupted brush border, loss of intercellular cohesion, hypertrophied tubular lumen, and hemolymph infiltration between cells were observed in some zones. Resorptive cells (R-cells) and vacuoles of blister-like cells (B-cells) of both species show a higher glycogen content at 35 psu than at 10 psu. At lower salinities, the cytoplasm of the different cell types evidence higher contents of carboxylated GPs in N. granulata and of su If at ed GPs in C. angulatus. At both salinities, and at the two pHs in N. granulata and at pH 5.6 in C. angulatus, the brush border, the vacuoles of B-cells and the peritrophic membrane show metachromasia. R-cell vacuoles and the cytoplasm of all cell types--except for the E-cells--at all salinities in both species show abundant lipid droplets. The results of the present study contribute significant data to the histophysiology of crustacean decapods, favoring the comprehension of the complex adjustment mechanisms facing saline stress in euryhaline crabs.

Surface ultrastructure of the gills of Odontesthes bonariensis (Valenciennes, 1835)(Teleostei: Atheriniformes) from a temperate shallow lake / Estudio ultraestructural de las branquias de Odontesthes bonariensis (Valenciennes, 1835) (Teleostei: Atheriniformes) de un lago somero templado

We studied the gill epithelium of juveniles of Odontesthes bonariensis using scanning electron microscopy with a particular interest in the relationship between cell morphology and the different functional aspects of gills. The specimens used were collected in Laguna de los Padres, Buenos Aires Province, Argentina. The gill filaments or primary lamellae are arranged in two rows along the convex outer face of the gill arch forming the anterior and posterior hemibranchiae. On its concave inner face, each arch supports two rows of gill rakers forming the pharyngeal region wall. The primary lamellae are lined by a stratified epithelium made up of polygonal cells with well-defined outlines and cell membrane microfolds. Several mucous and chloride cells were observed in the primary lamellae. The epithelial cells of the secondary lamellae have ill-defined outlines and few microfolds. Both the arches and gill rakers are covered by a stratified epithelium of polygonal cells, among which mucous cells and taste corpuscles are abundant. The ultrastructural analysis of gills of O. bonariensis contributes to the advancement of knowledge of the organ histophysiology and of future histopathological studies.

Se estudió el epitelio branquial de juveniles de Odontesthes bonariensis utilizando microscopía electrónica de barrido con un interés particular en la relación entre la morfología celular y los distintos aspectos funcionales de las branquias. Los ejemplares utilizados fueron colectados en la Laguna de los Padres, Provincia de Buenos Aires, Argentina. Los filamentos branquiales o laminillas primarias están dispuestos en dos filas a lo largo de la cara exterior convexa del arco branquial formando las hemibranquias anterior y posterior. En su cara interior cóncava, cada arco sostiene dos hileras de branquiespinas que forman la pared de la región faríngea. Las laminillas primarias están revestidas por un epitelio estratificado constituido por células poligonales con contornos bien definidos y micropliegues de la membrana celular. Se observaron numerosas células clorhídricas y mucosas en las laminillas primarias. Las células epiteliales de las laminillas secundarias tienen contornos poco definidos y escasos micropliegues. Tanto los arcos como las branquiespinas están cubiertos por un epitelio estratificado de células poligonales, entre las cuales hay abundantes células mucosas y corpúsculos gustativos. El análisis ultraestructural de las branquias de O. bonariensis proporciona información para avanzar en el conocimiento de la histofisiología del órgano y para realizar futuros estudios histopatológicos.

Morphology of the claw closer muscle in two estuarine crab species (Crustacea, Varunidae): an ultrastructural study.

We analyzed the ultrastructural features of the claw closer muscles in two estuarine crabs, Cyrtograpsus angulatus and Neohelice granulata, by transmission electron microscopy. Adult male crabs at intermolt stage were collected in the Mar Chiquita Coastal Lagoon (Buenos Aires, Argentina). The muscle fibers of both species showed evident striations, peripheral and intermyofibrillar nuclei, clefts in continuity with T and Z tubules, sarcoplasmic reticulum and T tubules forming dyads and triads usually located between the A and I bands, and mitochondria located mainly beneath the sarcolemma. Glycogen was observed as diffuse, small particles among myofilaments. The claw closer muscle of C. angulatus exhibited two fiber types: one with relatively fast-contracting fibers (shorter sarcomeres, myofilaments with an ordered arrangement, lineal Z discs, a well-developed sarcotubular system) and fatigue-resistant (numerous large mitochondria); and the other type, with slower-contracting fibers (longer sarcomeres, less orderly arranged myofilaments, wavy Z discs, a less developed sarcotubular system) and less resistant to fatigue (lower mitochondrial density). N. granulata showed only the slow, less resistant to fatigue type. The fibers less resistant to fatigue and more slowly contracting would presumably be used primarily for displays and agonistic interactions, whereas fast fibers with abundant mitochondria would be associated with continuous movements during feeding and grooming.

Preliminary Study of the Sonic Muscle of Micropogonias furnieri (Actinopterygii, Sciaenidae): Morfología e Histoquímica / Morphology and Histochemistry / Estudio Preliminar del Músculo del Sonido de Micropogonias furnieri (Actinopterygii, Sciaenidae)

The present study highlights the morphology of the Micropogonias furnieri sonic muscle by means of histochemical techniques, relating it to previous histochemical studies of the pectoral fin and myotomal muscles. In order to classify the muscle fibres, succinic dehydrogenase (SDH) for mitochondria, periodic acid Schiff (PAS) for glycogen, Sudan Black and Red for lipids and myosin adenosine triphosphatase (m-ATPase) pre-incubated at alkaline and acid pHs to visualize the contraction velocity, were used. The sonic fibres were smaller than the white myotomal and pectoral fibres, showed homogenous size and distribution and had features common to white fibres: negative to SDH and lipids, weakly positive to PAS and m-ATPase following acid pre-incubation, and positive to m-ATPase at alkaline pre-incubation. The morphology of the sonic muscle of M. furnieri showed no differences between sexes, probably due to the fact that the individuals were at the post spawning maturity stage. This similarity would indicate a similar sound production in both sexes, related to the "disturbance calls" of this species.

El presente trabajo estudia la morfología del músculo del sonido de Micropogonias furnieri por medio de técnicas histoquímicas y lo compara con trabajos previos de la histoquímica del músculo pectoral y miotomal. Con el fin de clasificar las fibras musculares se utilizaron distintas técnicas: succinato deshidrogenasa (SDH) para mitocondrias, ácido periódico Schiff (PAS) para evidenciar glucógeno, Sudan Black y Red para lípidos y miosin adenosina trifosfatasa (m-ATPasa) preincubada a pH alcalinos y ácidos para determinar la velocidad de contracción muscular. Las fibras del músculo del sonido son más pequeñas que las fibras blancas del músculo miotomal y pectoral, presentan un tamaño y distribución homogéneos, y tienen características histoquímicas de fibras blancas: son negativas para SDH y lípidos, débilmente positivas al PAS y a la técnica de m-ATPasa luego de preincubación ácida, y positivas a esta última técnica luego de preincubaciones alcalinas. La morfología del músculo del sonido de M. furnieri no mostró diferencias entre sexos, probablemente debido a que los individuos utilizados se encontraban en el estadio de post-desove. Esta similitud estaría relacionada con el tipo de sonido de "disturbio" que es emitido tanto por machos como por hembras durante todo el año.