A lectin histochemical study of gustatory (von Ebner's) glands of the horse tongue.

In the present work, gustatory glands (von Ebner's glands) of the horse tongue were examined by means of five peroxidase-conjugated lectins (PNA, DBA, SBA, UEA I, WGA), with and without prior sialidase digestion, in order to investigate the presence and distribution of carbohydrate residues in secretory cells and duct cells. The most intense staining of secretory cells was observed with PNA after pre-treatment with neuraminidase. This indicates that the terminal trisaccharide sequence sialic acid- (alpha 2-->3, 6) galactosyl (beta 1-->3) N-acetylgalactosamine is the most frequent oligosaccharide chain present in glycoproteins secreted by horse gustatory glands. Secretory cells also contained oligosaccharides with terminal alpha-N-acetylgalactosamine and N-acetylglucosamine, whereas fucose was found in only a few glandular cells. The apical cytoplasm of duct lining cells reacted with all the lectins except WGA.

Purification and some properties of a Mg(2+)-activated acid phosphatase from rat testis.

1. The acid phosphatase (AcPase, EC 3.1.3.2) IV from rat testicular tissue was purified to apparent homogeneity. 2. The enzyme displays a native molecular weight of 70 kDa determined on gel permeation chromatography on a Sephadex G-100 column and 68 kDa using linear 5-20% sucrose density gradient centrifugation. The subunit molecular weight on SDS-PAGE analysis is 67 kDa, suggesting that the enzyme is a monomeric protein. 3. The enzyme does not bind to Concanavaline A-Sepharose 4B column, indicating that it is not a glycoprotein. 4. The rat testis AcPase IV is a metal activated enzyme in which Mg2+ is the metal activating agent with a Ka = 0.88 x 10(-3) M. The Michaelis constant for p-nitrophenylphosphate, in the presence of saturating concentrations of Mg2+ ions, is 0.23 x 10(-3) M. 5. The enzyme preferentially hydrolyzes p-nitrophenylphosphate, phenylphosphate and ATP.

Characterization and fine-structural localization of actin- and fibronectin-like proteins in planaria (Dugesia lugubris s.l.).

Actin- and fibronectin-like proteins were characterized in the planarian, Dugesia lugubris s.l., by sodium dodecyl sulphate polyacrylamide gel electrophoresis and immunoblotting analysis using antisera to vertebrate actin and fibronectin. These antisera recognized protein bands of 42 kDa and 220 kDa, respectively. In addition, the immunohistochemical distribution of both actin- and fibronectin-like material was examined by using immuno-electron microscopy. Actin-like protein was localized in myofibrils in various differentiation stages, and in the peripheral cytoplasm and lamellipodia of cells that were migrating. The fibronectin-like component was associated with the extracellular matrix in the fibrillar structures and with the surface of the migrating cells. Our data suggest that similar cellular and molecular mechanisms are involved in cell-matrix interactions and in the morphogenesis of living organisms at different evolutionary levels.

Histological and histochemical studies on the chicken lingual glands.

1. Morphological and histochemical observations were done on the chicken anterior and posterior lingual glands. Histology, ultrastructure and glycoconjugate histochemistry were investigated by means of light and electron microscopy using staining specific for complex carbohydrates. 2. In the anterior lingual glands there are lateral and medial zones showing different morphological and tinctorial features. The secretory cells are typical mucous cells. 3. Histochemical reactions revealed the presence of acidic glycoconjugates with terminal sialic acid residues, and glycoconjugates vicinal diol and sulphate groupings in the secretory granules. 4. It is suggested that the main functions of lingual glands are the lubrication of boli and protection from micro-organisms.

The phorbol ester TPA dramatically inhibits planarian regeneration.

The effect of phorbol ester TPA (12-O-tetradecanoylphorbol-13-acetate) on head regeneration in decapitated planarians (Dugesia lugubris s.1.) has been studied. TPA-treatment soon after head amputation dramatically inhibited the regenerative process. Ultrastructural analysis revealed that the migration of fixed parenchymal cells (FPCs) to the wound area was strongly activated by TPA. FPCs interacted with various types of cells inducing lysis and phagocyting cell debris. The resulting fluid was removed through diaphanous protrusions appearing at the level of the wound zone. Moreover the close association of FPCs with neoblastlike cell clusters in the parenchyma indicated their possible role in the modulation of neoblast migration.

Immunoelectron microscopic localization of actin in migrating cells during planarian wound healing.

Wound repair in planarians is mainly characterized by two cell-migratory events involving the epidermis adjacent to the wound and its basement membrane. The first event is the migration of epidermal cells to cover the wound surface; the second one is the migration of newly differentiating replacement epidermal cells from the parenchyma to the epidermis. In addition to these events, migration of fixed parenchymal cells is observed during wound healing. All migrating cells were characterized by the presence of actin, as shown by the results obtained by means of indirect immunolocalization with fluorescent and electron microscopy. Migrating cells were heavily labeled with gold particles, which clustered at the level of cell-matrix and cell-cell contacts.

Ultracytochemical and biochemical evidence for guanylate cyclase in guinea pig testis.

Guanylate cyclase activity has been studied biochemically and cytochemically in guinea pig testis. The results of the biochemical assays indicate an equal distribution of this enzyme between the soluble and particulate fractions, which have a different sensitivity to adenosine triphosphate. The cytochemical results demonstrate that the reaction product of guanylate cyclase is detectable in the interstitial capillary endothelial cells and, in the seminiferous epithelium, mainly at the level of the adjacent surfaces of Sertoli and germ cells of the intermediate and adluminal compartments. Guanylate cyclase activity appears at the level of pachytene spermatocytes and persists throughout subsequent stages of development. The distribution in the seminiferous epithelium seems to indicate that guanylate cyclase is involved in the interrelationships between Sertoli and germ cells during gamete differentiation.

Immunohistochemical localization of cyclic AMP and ultrastructural demonstration of adenylate cyclase activity in the testis of Esox lucius at time of spermiation.

In the testis of Esox lucius at the time of spermiation, activity of cyclic adenosine 3',5'-monophosphate (cAMP) was immunocytochemically localized at the level of the Sertoli cells. In these cells adenylate cyclase activity was also ultracytochemically demonstrated by using adenylyl imidodiphosphate as a substrate. Reaction products of adenylate cyclase were primarily detectable on the basal and adluminal plasma membranes and on the surface of protrusions of the cell body into the lumen.

Cytochemical study on the distribution of adenylate cyclase in guinea pig testis.

The distribution of adenylate cyclase in testis, by means of a specific substrate adenylyl-imidodiphosphate (AMP-PNP), has been determined. Membrane-associated reaction products, indicative of adenylate cyclase activity, are localized by a complete cytochemical medium (containing 10 mM NaF) at the level of the basal compartment of the seminiferous epithelium, on the basal surface of Sertoli cells, and on adjacent plasma membranes of Sertoli cells and spermatogonial cells. At the level of the adluminal compartment, reaction products were found on adjacent plasma membranes of Sertoli cells and early or elongated spermatids. Adenylate cyclase reaction products are detectable by a basal incubation medium (without 10 mM NaF) only in the adluminal compartment on the spermatid plasma membranes.

Identification of lymphocytes extracted from bovine hemal nodes.

Lymphocyte populations extracted and separated from hemal nodes (HN) and peripheral blood of 20 male healthy bovines were characterized by surface markers and the rosette-forming test. After treatment with FITC- or peroxidase-conjugated antibovine IgG, about 22% of cells from HN and peripheral blood showed superficial fluorescence (B cells) and about 13% were able to form "E rosettes" (T cells) with sheep erythrocytes pre-treated with neuraminidase. Nearly equal percentages were obtained from peripheral blood lymphocytes. On the basis of the data shown, we can conclude that the function of HN is, at least in healthy animals, analogous with that of lymph nodes. In this study we also used the SEM to evaluate the possibility of classifying the lymphoid cells as B and T lymphocytes on the basis of their surface morphology. Some authors have stated that B and T lymphocytes have a rough and smooth surface, respectively. In our experience, however, because of cells with a moderate number of surface microvilli exist, this method used on its own is not suitable for lymphocyte identification.