New findings on 3-D microanatomy of cellular structures in human tissues and organs. An HRSEM study.

We present here findings obtained on a large number of human tissues over a period of more than ten years, by our modification of the Osmium maceration method for high resolution scanning electron microscopy (HRSEM). Data are documented by original pictures which illustrate both some 3-D intracellular features not previously shown in human tissues, and results obtained in our current studies on mitochondrial morphology and on the secretory process of salivary glands. We have demonstrated that mitochondria of cells of practically all human tissues and organs have usually tubular cristae, and that even the cristae that look lamellar are joined to the inner mitochondrial membrane by tubular connexions similar to the crista junctions later seen by electron tomography.

Cytomorphological study on human submandibular gland following treatment with secretagogue drugs.

Using specimens of human submandibular glands, we have investigated in vitro the morphological modifications induced by clozapine, a dibenzodiazepine derivative that is used in psychotic patients and that provokes hypersalivation, a side-effect of therapy. The effects of the drug, used alone or in combination with carbachol, have been compared with those observed after treatment with drugs acting on specific receptors. To quantify the response to stimulation, we have calculated (with statistical methods) the number of microvilli and microbuds (corresponding to pits seen in images obtained by transmission electron microscopy) per square micrometre of the cytoplasmic surface of the intercellular canaliculi luminal membrane in images obtained by high-resolution scanning electron microscopy.

Morphological alterations induced by cytochalasin D on serous cells of human submandibular gland in basal and stimulated conditions.

Cytochalasin D (CD) is a fungal toxin which binds to the faster growing end of actin microfilament and inhibits actin polymerization. By an in vitro incubation system of slices of human submandibular glands obtained at surgery, we investigated by light microscope (LM), transmission electron microscope (TEM), and high resolution scanning electron microscope (HRSEM) the morphological changes caused by CD on serous cells. We studied the effects of the drug on secretory events induced by isoproterenol (I) and carbachol (C).

Serous and mucous cells of human submandibular salivary gland stimulated in vitro by isoproterenol, carbachol and clozapine: an LM, TEM, and HRSEM Study.

We have investigated by LM, TEM, and HRSEM the effects of D,L-isoproterenol (beta-adrenergic agent), carbachol (muscarinic agent) and clozapine on biopsy specimens of human submandibular gland stimulated in vitro in an inorganic oxygenated medium. Clozapine is a dibenzodiazepine derivative used in psychotic patients that provokes hypersalivation, a displeasing side effect that often causes discontinuance of therapy. Our findings demonstrate that clozapine acts on salivary mucous and seromucous (serous) cells of the gland as a muscarinic agonist.

Morphofunctional studies on human labial salivary glands.

In this study, the first experimental investigation carried out at the ultrastructural level on mucous cells of human salivary glands, we have examined by light microscopy (LM), transmission electron microscopy (TEM), high resolution scanning electron microscopy (HRSEM), the secretory response of labial glands stimulated in vitro by the beta-adrenergic agent, D,L isoproterenol, and by the muscarinic agent carbachol. For comparison we have used identical methods to study samples of mixed portions of human submandibular glands. Morphological findings obtained here on both submandibular and labial glands mucous cells demonstrate that mucous droplets are released solely by muscarinic stimulation, and that cytological events occurring during secretory discharge are similar to those described by others, using TEM, on stimulated mucous cells of rat sublingual glands.

Junctional complex revisited by high-resolution scanning electron microscopy.

The present study correlates the ultrastructural morphology of junctional complexes as revealed by transmission electron microscopy (TEM) with that observed by high-resolution scanning electron microscopy (HRSEM), thanks to a new modification of the osmium tetroxide maceration technique. The removal of all cytoplasmic organelles by this technique allows the inspection of the inner side of the plasmalemma. With this treatment, a continuous band of tightly packed particles is observed at the most apical portion of lateral membranes.

Fluid and amylase secretion by perfused parotid gland: physio-morphological approach.

Whole gland perfusion technique was applied to rat parotid glands to assess whether amylase affects fluid secretion. Control perfusion without any secretagogue evoked no spontaneous secretion. Carbachol (CCh 1 microM) induced both amylase and fluid secretion with distinctive kinetics.

Cell biology of human salivary secretion.

We treated surgical specimens of human parotid and submandibular glands in vitro to manipulate the receptor-signaling cascade pharmacologically and analyzed cellular responses by light microscopy on epoxy embedded sections. Treatment of specimens with the b-agonist, isoproterenol, and with the second messenger analog, dibutyryl cyclic AMP, stimulated serous acinar cells to engage in exocytosis and degranulation. The muscarinic agonist, carbachol, and the calcium ionophore, A23187, on the other hand, elicited formation of "vacuoles" in the cytoplasm of serous acinar cells.

A high resolution sem study of human minor salivary glands.

By SEM we have investigated the human minor salivary glands using the NaOH method for the visualization of endpieces and myoepithelial cells, and the osmium maceration technique that reveals membranous intracellular structures. With the former method all minor glands, including the posterior deep (Ebner's) lingual glands, consist of tubules sometimes dilated into alveoli, while true acini of the kind observed in human major salivary glands, are absent. Tubules of the posterior deep lingual gland exhibit stellate myoepitelial cells that leave a substantial part of the secretory cells uncovered.

A scanning and transmission electron microscope study of the human minor salivary glands.

All human minor salivary glands, apart from the posterior deep lingual (von Ebner's) glands which were serous, contained a minor population of seromucous cells that increased from palatine and posterior superficial lingual (Weber's) to labial, anterior lingual (Blandin and Nuhn's) and buccal glands, in that order. Unlike the predominant mucous cells, whose structure was uniform, serous and seromucous cells exhibited, in each gland, peculiar cytological and cytoarchitectural characters.

Cytoskeletal regulation of human salivary secretion studied by high resolution electron microscopy and confocal laser microscopy.

To study the cell regulation mechanisms of human salivary secretion, surgical specimens of human parotid and submandibular glands were treated in vitro with isoproterenol (beta-agonist), carbachol (muscarinic agonist), and cytochalasin D (microfilament disruptive agent), and morphological changes occurring in serous acinar cells were observed. Control acinar cells treated without secretagogues exhibited only occasional examples of exocytosis. Microfilaments, revealed by transmission electron microscopy (TEM) and confocal laser scanning microscopy (CLSM) of F-actin fluorescence stained by rhodamine-phalloidin, were localized underneath the luminal membrane to separate the secretory granules from the luminal membrane.

Human salivary gland parenchymal cells seen by SEM from the cytoplasmic side using a new osmium maceration method.

By removing all or most organelles, we have exposed the cytoplasmic side of the plasmalemma and its specializations in serous cells and in cells of striated and excretory ducts of human major salivary glands. The areas of plasmalemma located beneath the lumen and those bordering the intercellular canaliculi are covered by evenly distributed particles arranged in a continuous band and, below it, in regularly spaced clusters. A similar pattern of particles is seen on the internal aspects of the juxtaluminal plasmalemma of cells of both striated and excretory ducts.

Historical note: Giuseppe Oronzo Giannuzzi and the discovery of the serous demilunes and of the intercellular canaliculi.

The tragic life and scientific achievements of Giuseppe Oronzo Giannuzzi are briefly outlined. Particular attention is focused on the discovery, made in Karl Ludwig's laboratory, of the serous demilunes and of the intercellular canaliculi of salivary glands.

Exocytosis in human salivary glands visualized by high-resolution scanning electron microscopy.

The luminal membrane of salivary acinar cells creates a specialized cell surface area that accepts exocytosis and undergoes dynamic changes during secretion. These changes were visualized three-dimensionally from both the inside and outside of the cell in human parotid and submandibular glands, by application of in vitro secretory stimulation and then of OsO4 maceration to remove cytoplasmic organelles by varying degrees. In control glands treated without secretagogues, the luminal surface of serous acinar cells bore well-developed microvilli with only an occasional incidence of exocytotic profiles.

Further data on intracellular structures of human salivary glands. A SEM study.

Specimens of human salivary glands have been studied by our modification of the AODO maceration method which, carried out on sections of controlled thickness, allows the analytical study of human bioptical material. Lately, our technique has been further improved and simplified by omitting the treatment with dimethylsulfoxide and by using osmium-ferrocyanide as secondary fixative. Following maceration with diluted OsO4, some of the sections also were shaken for 10-15 min with a rotating agitator.

Niels Stensen (Niccolò Stenone) and his first scientific offspring: the salivary glands.

Niels Stensen's (Niccolò Stenone) life and scientific achievements are briefly outlined and discussed. Particular attention is focused on his contribution to the understanding of salivary and exocrine secretions.

The main excretory duct (Stensen's) of the human parotid gland: a transmission and scanning electron microscope study.

The epithelial cells of the human parotid main excretory duct (Stensen) were studied by transmission (TEM) and scanning (SEM) electron microscopy through a variety of procedures that allowed the visualization of their three-dimensional microanatomy. Stensen's duct in humans is lined, in its distal portion, with a pseudostratified epithelium with tall principal cells and smaller basal cells, while the epithelium becomes progressively stratified cylindrically toward the oral stoma. Goblet cells are scattered among the other epithelial cells.

3D-structure of cells of human salivary glands as seen by SEM.

To demonstrate by SEM the topography and cytoarchitecture of the different parenchymal components of human salivary glands, we have employed a number of techniques that allow either the exposure of internal and lateral cell surfaces or, following the removal of connective tissue, the visualization of endpieces, ducts, and myoepithelial cells. Serous glands consist of indented acini attached to the ducts in a grape-like fashion, whereas mucous and mixed glands are made up of smooth tubuli. Myoepithelial cells (mecs), which are abundant on the surfaces of acini, tubuli, and intercalated ducts, are sparse on striated ducts.

3D microanatomy of human parotid gland.

We have performed a scanning electron microscope study on human parotid gland. By using a variety of techniques of maceration and digestion we have shown the 3D morphology of cells and of isolated endpieces.

Fine structure of excretory ducts of human salivary glands.

Excretory ducts of human major salivary glands are lined by an epithelium consisting of principal cells and by a discontinuous row of basal cells. The principal cells are tall and columnar with mitochondria, large lipofuscin granules and a central nucleus. Just beneath the plasmalemma bordering the lumen, their cytoplasm contains a number of small granules and vesicles similar to those observed in cells of striated ducts.

Ultrastructure of human bulbourethral glands and of their main excretory ducts.

The endpieces of human bulbourethral (BU) glands, studied with SEM after removal of the connective tissue, consist of short, coiled tubules often dilated into alveoli. Immunohistochemical studies at the EM level have shown that the mucous cells of these glands have mucous droplets, which react to blood group antigens, suggesting that BU glands participate in the secretion of these antigens into the seminal plasma. The main excretory duct is lined by a stratified columnar epithelium consisting of six-seven cellular layers.

Ultrastructural observations on human sublingual gland.

That part of the human sublingual gland that corresponds in morphology to the conventional description of this organ presented in most histology texts (probably the major sublingual gland, in contradistinction to the aggregated small glands that compose the minor sublingual glands) was studied by electron microscopy. The gland is mixed, with slightly more mucous elements than seromucous ones. The mucous cells are arranged in tubules that usually are capped by seromucous demilunes.

The human bulbo-urethral glands. A transmission electron microscopy and scanning electron microscopy study.

This paper investigates the ultrastructure of human bulbo-urethral glands using specimens obtained at surgery. The tubulo-alveolar endpieces of these glands are lined by typical mucous cells in different stages of the secretory cycle. The most interesting features of their cytoplasm are membrane-bounded bodies with a filamentous texture that usually fuse with the mucous droplets before they discharge into the lumen.

Ciliated cells in the main excretory duct of the submandibular gland in obstructive sialadenitis: a SEM and TEM study.

The epithelium of the main excretory duct of the submandibular gland in five cases of obstructive sialadenitis was studied by TEM and SEM. In three cases we found a considerable increase in goblet and ciliated cells. As well as normal cilia, different kinds of ciliary anomalies, especially compound cilia, were noticed.