Histochemical and morphometric analyses of the musculature of the forelimb of the subterranean rodent Ctenomys talarum (Octodontoidea).

Histochemical and morphometric analyses were performed to characterize the fibre-type composition of two forelimb muscles of the South American subterranean rodent Ctenomys talarum. The studied muscles were the triceps lateralis, an extensor of the elbow, and the teres major, a flexor of the shoulder. It was found that these muscles had an elevated proportion of fast oxidative-glycolytic (FOG) fibres, and lower proportions of slow oxidative (SO) and fast glycolytic (FG) fibres. This composition probably qualifies the teres major and triceps muscles to perform the sustained effort required in tunnelling excavation. The results were discussed considering published data on fibre-type composition of mammals having different modes of life and digging behaviour. We here suggest that C. talarum has the potential of generating forces linked to rapid, powerful movements during sustained periods by means of an elevated proportion of FOG fibres, together with osteological changes that result in a great mechanical advantage of the forelimb muscles.

Morphology, morphometry, histochemistry and lectin histochemistry of the vagina of the plains viscacha (Lagostomus maximus).

The aim of the present work was to describe the morphology of the vagina in Lagostomus maximus and to characterize its epithelial cells using morphometric and histochemical techniques (variations of PAS, Alcian blue and lectin histochemistry). Thirty-five sexually mature adult females were captured in their natural environment during four periods of the year and their genital organs were dissected. The vaginal wall of the viscacha has three tunics: mucosa, muscularis and adventitia or serosa according to the region. The epithelium is stratified in both cranial and caudal regions, but its characteristics vary depending on the physiological state. In anestrous, nonpregnant females have a stratified epithelium of two to three cellular layers with columnar PAS-positive superficial mucous cells. During the follicular phase, the epithelium of the vagina is stratified squamous and cornified. Females at early, middle and term pregnancy have a columnar stratified epithelium with mucous cells. Glycoproteins in the mucous cells were detected using PAS, PA*S, KOH/PA*/BH/PAS; and Alcian blue, pH 0.5, pH 1, pH 2.5 and 0.006 M). Lectin histochemistry showed that UEA-I and RCA-1 lectins reacted strongly or moderately with epithelial cells in all stages analyzed. These results indicate the presence of L-fucose and ß-galactose. Binding with other lectins was variable.

Glycoproteins histochemistry of the gills of Odontesthes bonariensis (Teleostei, Atherinopsidae).

The histochemistry of glycoproteins (GP) in the mucous cells of the gills of the silverside Odontesthes bonariensis was identified with: (1) oxidizable vicinal diols; (2) sialic acid and some of their chain variants, carbon 7 ((7) C), carbon 8 ((8) C) or carbon 9 ((9) C); (3) sialic acid residues without O-acyl substitution and with O-acyl substitution at (7) C, (8) C or (9) C; (4) carboxyl groups and (5) sulphate groups. A battery of seven biotinylated lectins allowed GPs sugar residues to be distinguished. Mucous cells showed the presence of neutral, sulphated and sialylated GPs. Dolichos biflorus agglutinin (DBA) and Glycine max agglutinin (SBA) showed strong positive staining; Arachis hypogaea agglutinin (PNA), Ricinus communis agglutinin-I (RCA-I) and Triticum vulgaris agglutinin (WGA) showed moderate staining, while Ulex europaeus agglutinin-I (UEA-I) was completely negative.

Pectoral fins of Micropogonias furnieri: a histochemical and ultrastructural study.

The myotomal fibres of the pectoral fins of white croaker (Micropogonias furnieri) have been studied using histochemical techniques and transmission electron microscopy (TEM). Succinic dehydrogenase (SDH) for mitochondria, periodic acid Schiff (PAS) for glycogen, Sudan Black for lipids and myosin-adenosintriphosphatase (mATPase) pre-incubated at alkaline and acid pHs were used to visualize the contraction velocity. Three zones were determined: superficial (SZ), medium (MZ) and deep (DZ). Staining for SDH, PAS and Sudan Black was positive only in the SZ. The level of alkaline mATPase was the highest in fibres from the DZ, intermediate in the MZ and low in the SZ; at an acid pH, the reverse was obtained. Fibres from the SZ were small with large quantities of subsarcolemmal mitochondria, scarce intermyofibrilar mitochondria and a well-developed sarcoplasmic reticulum; the myofibrils displayed a polygonal distribution along the entire length of the fibre. Fibres in the MZ were larger than those in the SZ, the myofibrils were densely packed, mitochondria prevailed under the sarcolemma and the sarcoplasmic reticulum was not abundant. Fibres from the DZ were the largest, with ribbon-shaped myofibrils and scarce mitochondria. The intercellular space was abundant and nervous endings were frequently observed.

Morphological and histochemical characteristics of the epithelium of ovarian lamellae of Genypterus blacodes (Schneider, 1801).

The physiological significance of the glycoproteins (GPs) secreted by the epithelium of ovarian lamellae is discussed in reference to the reproductive biology of G. blacodes. Histochemical procedures for localising and characterising GPs were used to determine the cytoplasmic components of cells of the epithelium that covers the ovarian lamellae of pink cuskeel, Genypterus blacodes (Schneider, 1801) (Pisces, Ophidiidae), during spawning. This species is one of the most valuable demersal fish resources in the Argentine Sea, mainly due its large size and flesh quality. GPs with oxidizable vicinal diol groups, sialic acid with or without O-acyl substituents, O-acyl sugars, neutral sugars and GPs with carboxyl and sulphate groups were detected. Light microscope examination showed morphological changes in the epithelium of ovarian lamellae during the spawning season, associated with a secretory activity of mucus. Optical density studies revealed the presence of polyploid cells encompassing those morphological changes. Results of the present study suggest that the epithelium of ovarian lamellae of G. blacodes performs a secretory role, which is intensified during ovarian maturity, suggesting that G. blacodes could release masses of eggs enveloped in mucus.

The mucosa of the digestive tract in Micropogonias furnieri: a light and electron microscope approach.

The histomorphological aspects as well as the histochemical content and distribution of glycoproteins (GPs) in the mucosa of the digestive tract of the white croaker Micropogonias furnieri were studied. The buccopharyngeal cavity and the esophagous showed a squamous stratified epithelium with mucous cells. The stomach presented three portions: cardias, fundus and pylorus. Tubular glands formed by a single type of gland cell were located along the cardias and fundus. Histochemical tests showed that the buccopharyngeal cavity and the esophagous presented the largest amount of the different types of mucosubstances. Both organs showed abundant secretory mucous cells that synthesize large quantities of neutral, sulphated and sialylated GPs. The surface epithelium in the cardias and fundus synthesized and secreted scarce sialylated and neutral GPs whereas the secretions of the apical surface were abundant. The pylorus secreted large amounts of neutral as well as sulphated and sialylated GPs. Gland cells secreted neutral GPs. The ultrastructural features of the gut cells were quite similar to those of other teleosts. The buccopharyngeal cavity and the esophagous surface epithelial cells, identified by their superficial localization, were characterized by cytoplasmic vesicles of different size. Abundant goblet cells with secretory mucous granules were also present. Gastric glands in the stomach contained just one form of cell with a fine structure similar to cells that secrete pepsinogen.

Histology and histochemistry of the pharyngeal cavity and oesophagus of the silverside Odontesthes bonariensis (Cuvier and Valenciennes).

The histology of the pharyngeal cavity and oesophagus of the freshwater 'silverside'Odontesthes bonariensis (Cuvier and Valenciennes) and the characteristics of their mucous cells were investigated. The histological characterization of its digestive wall revealed that the mucosa is thrown with longitudinal folds. The epithelium covering the folds was stratified with abundant mucous cells and gustative corpuscles, which are lacking in the oesophagus. The muscularis mucosa was absent. The submucosa presented the compactum stratum. The muscularis was organized in longitudinal and circular layers of muscular striated fibres. The serosa with a flat epithelium was located only in the oesophagus. Using histochemical procedures including methods for localization and characterization of glycoproteins (GPs), no differences were detected between the mucous cells contents of the pharyngeal cavity and those of the oesophagus. The mucous cells showed a weak periodic acid-Schiff (PAS) positive reaction in their content. The reactions for the differential analysis of sialic acids from GPs are feeble for periodic acid-Schiff at low temperature and low pH (PA*S) and KOH/PA*S and strong for periodic acid/borohydride/KOH/PAS (PA/Bh/KOH/PAS) and KOH/PA*/Bh/PAS revealing the scarce presence of C7 or C9 substituted and non-substituted sialic acids and the abundance of C7, C8 substituted sialic acids, O-acyl sugars and neutral sugars respectively. The results suggest that the pharyngeal cavity with the gustative corpuscles would induce the gastric secretion whereas the oesophagus is mainly involved in the transport of the food bolus to the stomach with the aid of abundant secretion of mucus. GPs secreted on the surface of the mucous cells, likely related to environmental conditions, would be involved in the lubrication, protection against abrasion and inhibition of microorganism proliferation.

Ultrastructure and histochemical study of glycoconjugates in the gills of the white croaker (Micropogonias furnieri).

The ultrastructure of the primary and secondary lamellae of gills was investigated in a marine teleost, the white croaker. The following cells were identified and briefly described: pavement cells, mucous cells, mitochondria-rich cells and rodlet cells. These cell types are present throughout the length of the lamellae. They are studied by means of a series of carbohydrate histochemical methods, including lectin procedures. Neutral sugars and substituted sialic acid were detected by means of periodic acid-borohydride reduction-saponification-periodic acid Schiff reaction (PA/Bh/KOH/PAS), saponification-selective periodic acid Schiff reaction (KOH/PA*S) and saponification-selective periodic acid-borohydride reduction-periodic acid Schiff reaction (KOH/PA*/Bh/PAS) histochemical techniques. A battery of seven lectins was used to study binding on tissue sections at the light microscopic level to characterize glycoconjugates in gills. The reaction to Canavalia ensiformis agglutinin (Con-A), Triticum vulgaris agglutinin (WGA), and Ricinus cummunis agglutinin-1 (RCA-1) was weak in pavement cells; unlike Con-A, the reaction to WGA and RCA-1 was more intense in mucous cells. Arachis hypogaea agglutinin (PNA) lectin showed a strong reaction in mucous cells. Ulex europaens agglutinin-1 (UEA-1) lectin was negative in all cell types. The lectin pattern was similar for both primary and secondary lamellae, except for PNA reaction, which was weak in the pavement cells of the secondary lamella and negative in the pavement cells of the primary lamella.

Morphological and histochemical characterization of the mucosa of the digestive tract in Engraulis anchoita.

The histomorphological aspects as well as the histochemical content and distribution of glycoproteins (GPs) in the mucosa of the digestive tract of the anchovy Engraulis anchoita were studied. The buccopharyngeal cavity is lined by a squamous stratified epithelium with mucous superficial cells; the oesophagus shows two zones, cranial with a squamous stratified epithelium with mucous superficial cells and caudal with a columnar secretory epithelium. Finally, the stomach presents both the cranial and pyloric portion lined with a simple columnar epithelium. Tubular branched glands, formed by a single type of glandular cell, located along the stomach, are more numerous in the cranial portion. The GPs were identified with (1) oxidizable vicinal diols; (2) sialic acids and some of their chain variants, C7 or C9; (3) sialic acid residues with O-acyl substitution at C7 or C8; (4) carboxyl groups and (5) sulphate groups. Histochemical tests showed that the buccopharyngeal cavity presented the largest amount of the different types of mucosubstances. Epithelial secretory cells were found in the oesophagus, which synthesized a large quantity of sialosulphoglycoproteins likely to be related to a protective role. The surface epithelium of the stomach synthesizes and secretes acid and neutral GPs, probably related to the movement of fluids and to the absorption of easily digested substrates, respectively. Although great differences exist between different species, in E. anchoita as in other fish species, the wall of the digestive tract is composed of the four layers classically described for vertebrates. The GPs secreted by the epithelial cells are suggested to be important for the protection and inhibition of microorganisms. In addition, they are involved in enzymatic digestion of food, absorptive functions and lubrication of the alimentary tract.

Mucous cells in micropogonias furnieri gills: histochemistry and ultrastructure.

The characteristics of the mucous cells located in the gills of the fish Micropogonias furnieri were investigated. Using histochemical procedures that included methods for localization and characterization of glycoproteins (GPs), no differences were detected between the mucous cell contents of the primary and secondary lamellae. The GPs were identified with (a) oxidizable vecinal diols; (b) sialic acids and some of their chain variants, C7 or C9; (c) carboxyl groups and (d) sulphate groups. The electron microscope showed large mucous globules of different electro densities from mucous cells located deep in the epithelium between the other epithelial cells; the release of mucus by exocytosis was observed. GPs secreted on the surface of the mucous cells was suggested to be important for the lubrication, protection and inhibition of microorganisms. It is possibility that GPs could have similar roles in Micropogonias furnieri gills.

Characterization of the swimming muscle of the anchovy Engraulis anchoita (Hubbs and Martini 1935).

The lateral muscle characteristics of individual female anchovies (Engraulis anchoita) were studied by histochemical and ultrastructural techniques. Red, pink and white fibres were discerned on the basis of the activity of the metabolic enzymes succinic dehydrogenase, phosphorylase and myofibrilar ATPase, and the usage of Sudan and periodic-acid Schiff staining techniques. Red fibres were located at the surface, white fibres deep inside and pink fibres in between. The red fibres appeared flattened or ribbon-shaped in transverse section and they were located in rows separated by myosepta of connective tissue. The red and pink fibres were multi-innervated whereas the white fibres were terminally innervated. The relative proportion of the red muscle increased toward the caudal region, the white muscle diminished toward that region, whereas the proportion of pink muscle did not vary according to region, being scarce throughout. The distribution of capillaries was also investigated, and as a result, the vascular supply proved to be scarce in both white and pink muscle.

Lateral musculature in the whitemouth croaker (Micropogonias furnieri): its characterization with respect to different gonadal conditions.

Histochemical and ultrastructural studies were performed on the lateral musculature from individual female whitemouth croaker, Micropogonias furnieri, at the anterior, medium and posterior regions. Based upon histochemical myosin-ATPase (m-ATPase) determination, diverse types of red, pink and white fibres were discerned. Red muscle had abundant mitochondria and stained intensely for aerobic enzymes, white muscle scarcely stained for the same enzymes and pink muscle responded in an intermediate manner. Both white and pink muscle had few mitochondria. The relative proportion of red muscle increased towards the caudal region; pink muscle diminished towards this region and white muscle modified its proportion only in the anterior region. m-ATPase activity showed differences in relation to the gonadal condition along the body, particularly in the white fibres at the anterior and medium regions.

Characterization of lateral musculature in the striped weakfish (Cynoscion striatus Cuvier).

Histochemical and ultrastructural studies were performed on lateral musculature from individual female weakfish (Cynoscion striatus Cuvier). Based upon SDH, PhR, Sudan and myosin-ATPase determinations, different kinds of red, pink and white fibres were discerned at the anterior, medium and posterior regions. Levels of glycolytic activity increased from red to white fibres, being intermediate in the pink ones. Contrarily, oxidative activity decreased from red to pink and white fibres. Histochemical stain showed a complex distribution of fibre types in each layer. A considerable variation in size and distribution was also found among fibres.

Lipid accumulation inRhodotorula glutinis on sugar cane molasses in single-stage continuous culture.

Microbial lipids produced byRhodotorula glutinis grown in continuous culture with molasses under nitrogen-limiting conditions were evaluated and the effects of growth rate on fatty acid composition were studied. As the growth rate decreased, cell biomass, lipid content and lipid yield gradually increased. The maximum lipid content recorded was 39% (w/w) of dry cell biomass at a dilution rate of 0.04 h(-1). The growth rate also affected fatty acid composition: oleic acid decreased with decreasing growth rate while stearic acid increased.