Morphological and histochemical characterization of the secretory epithelium in the canine lacrimal gland.

In the present study, the expression of secretory components and vesicular transport proteins in the canine lacrimal gland was examined and morphometric analysis was performed. The secretory epithelium consists of two types of secretory cells with different morphological features. The secretory cells constituting acinar units (type A cells) exhibited higher levels of glycoconjugates, including ß-GlcNAc, than the other cell type constituting tubular units (type T cells). Immunoblot analysis revealed that antimicrobial proteins, such as lysozyme, lactoferrin and lactoperoxidase, Rab proteins (Rab3d, Rab27a and Rab27b) and soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor (SNARE) proteins (VAMP2, VAMP4, VAMP8, syntaxin-1, syntaxin-4 and syntaxin-6), were expressed at various levels. We immunohistochemically demonstrated that the expression patterns of lysozyme, lactoferrin, Raba27a, Rab27b, VAMP4, VAMP8 and syntaxin-6 differed depending on the secretory cell type. Additionally, in type T cells, VAMP4 was confined to a subpopulation of secretory granules, while VAMP8 was detected in almost all of them. The present study displayed the morphological and histochemical characteristics of the secretory epithelium in the canine lacrimal gland. These findings will help elucidate the species-specific properties of this gland.

Development of vasoactive intestinal polypeptide-immunoreactive nerves in the major cerebral arteries of the quail anterior circulation.

Development of cerebral perivascular nerves immunoreactive for vasoactive intestinal polypeptide (VIP) was investigated in the Japanese quails, using immunohistochemistry and quantitative analysis. VIP-immunoreactive (VIP-IR) nerves supplying the anterior circulation appeared on the cerebral carotid artery (CCA) at embryonic day 10 and on the cerebroethmoidal artery (CEA) after hatching. Nerves from the CCA increased greatly in number and spread progressively during successive embryonic stages, while those from the CEA were sparse all through the post-hatching stages, mostly remained limited to this vessel wall. The distribution of VIP-IR nerves to the respective major arteries of the anterior circulation from the two vascular routes was basically similar among post-hatching day (P) 15, P20, P30 and P50. Likewise, no clear statistical difference was observed with regard to the nerve density of the corresponding arteries in the four age groups. These findings suggest that VIP-IR innervation of the quail anterior circulation usually attains its mature pattern at the third week after hatching.

Lectin histochemistry of the endothelium of blood vessels in the mammalian integument, with remarks on the endothelial glycocalyx and blood vessel system nomenclature.

Based on sensitive light and electron microscopical lectin histochemistry, the distribution of saccharide residues is demonstrated in endothelial cells and/or the walls of integumental blood vessels of domesticated and wild mammals. In addition, the nomenclature of the blood vessel system in the skin is reviewed, and modified according to a generalized mammalian approach. Our comparative attempt demonstrated three (upper, mid-dermal, and dermal) plexus or retia in the integument of mammals of important systematic groups. The findings highlight a specific spectrum of terminal sugars in the endothelial cells and their glycocalyx and/or the blood vessel wall as related to the vessel retia and plexus present. The subepidermal blood vessel system, the capillary loops in particular, was marked by alpha-Man/alpha-Glc, alpha-D-GalNAc, beta-D-Gal/beta-D-GalNAc, and NANA-alpha(2,6)-GalNAc; the mid-dermal system by alpha-Man/alpha-Glc, and alpha-D-Gal/alpha-D-GalNAc; and the deep dermal system by alpha-Man/alpha-Glc, alpha-D-GalNAc, alpha-Gal, and beta-D-Gal/beta-D-GalNAc moieties. The results obtained are discussed from a comparative point of view, also with regard to certain basic functions of the endothelial cells and their glycocalyx.

Immunocytochemical localization of lysozyme and beta-defensin in the apocrine glands of the equine scrotum.

The present study revealed in detail the subcellular localization of lysozyme and beta-defensin in the apocrine glands of the equine scrotal skin, a specific body region. The apocrine glandular cells were equipped with a varying number of secretory granules, a well-developed Golgi apparatus and abundant cisternae of the rough endoplasmic reticulum within their cytoplasm. In these cells, reactive gold particles representing lysozyme were detectable in the secretory granules as well as the Golgi apparatus and elements of the rough endoplasmic reticulum. Additionally, the antimicrobial peptide group of beta-defensin was also localized in the above-mentioned ultrastructures of the secretory cells. The presence and secretion of such substances that may serve as a non-specific defense against microorganisms are suggestive of the protective effect of the secretory production elaborated by the apocrine glands.

Ultrastructure and immunohistochemical characterization of proteins concerned with the secretory machinery in goat ceruminous glands.

The expression of soluble N-ethyl-maleimide sensitive fusion attachment protein receptor (SNARE) proteins in apocrine glands has not been fully elucidated. In addition to performing ultrastructural observation of the ceruminous glands in goats, our study focuses on the demonstration of ß-defensins, SNARE proteins and Rab3D in these glands with the use of immunohistochemical methods. The secretory cells were equipped with two types of vesicles, Golgi apparatus and abundant rough endoplasmic reticulum (ER). Additionally, in some of them, the characteristic concentric structures composed of rough ER were observed in their circum- and infranuclear parts. The expression of phosphorylated inositol requiring enzyme 1a was also detected. These findings may indicate their ability to produce numerous secretory proteins and the maintenance of homeostasis in the glandular cells. Furthermore, ß-defensins were demonstrated as products of the ceruminous glands. The present investigation also revealed the presence of SNARE proteins and Rab3D. It is suggested that these proteins are concerned with the secretory machinery of this gland type.

Canine Salivary Glands: Analysis of Rab and SNARE Protein Expression and SNARE Complex Formation With Diverse Tissue Properties.

The comparative structure and expression of salivary components and vesicular transport proteins in the canine major salivary glands were investigated. Histochemical analysis revealed that the morphology of the five major salivary glands-parotid, submandibular, polystomatic sublingual, monostomatic sublingual, and zygomatic glands-was greatly diverse. Immunoblot analysis revealed that expression levels of α-amylase and antimicrobial proteins, such as lysozyme, lactoperoxidase, and lactoferrin, differed among the different glands. Similarly, Rab proteins (Rab3d, Rab11a, Rab11b, Rab27a, and Rab27b) and soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor (SNARE) proteins VAMP4, VAMP8, syntaxin-2, syntaxin-3, syntaxin-4, and syntaxin-6 were expressed at various levels in individual glands. mmunohistochemistry of Rab3d, Rab11b, Rab27b, VAMP4, VAMP8, syntaxin-4, and syntaxin-6 revealed their predominant expression in serous acinar cells, demilunes, and ductal cells. The VAMP4/syntaxin-6 SNARE complex, which is thought to be involved in the maturation of secretory granules in the Golgi field, was found more predominantly in the monostomatic sublingual gland than in the parotid gland. These results suggest that protein expression profiles in canine salivary glands differ among individual glands and reflect the properties of their specialized functions.

Morphology and glycoconjugate histochemistry of the eccrine glands in the snout skin of the North American raccoon (Procyon lotor).

The eccrine nasolabial glands were found in the hypodermis of the nasal plane in the North American raccoon (Procyon lotor). In addition to light and electron microscopic observations, the distribution and selectivity of complex glycoconjugates in the eccrine tubular glands of the raccoon snout skin were studied using various histochemical methods, particularly lectin staining. The secretory epithelium and the luminal secretions exhibited high amounts of glycoconjugates with various saccharide residues (alpha-D: -mannose, alpha-L: -fucose, beta-D: -galactose, beta-N-acetyl-D: -glucosamine, sialic acid). The excretory duct cells also showed positive reactions with most of the histochemical methods applied. The results are discussed with regard to possible functions of the glandular secretions. The complex glycoconjugates that are produced by the eccrine nasolabial glands may be related to moistening of the skin surface as well as protecting the epidermis against physical damage or microbial contamination. This is the first report on the glands in the snout skin of carnivores.

Histochemical localization of complex carbohydrates in the nasolabial glands of the Japanese deer (Cervus nippon yakushimae).

The localization of complex glycoconjugates in the nasolabial glands of the Japanese deer (C. nippon yakushimae) was studied using various histochemical methods, including lectin histochemistry, viewed using both light and electron microscopy. The secretory epithelium and luminal secretion of the deer nasolabial glands exhibited neutral and acidic glycoconjugates with different saccharide residues (alpha-D-mannose, alpha-N-acetyl-D-galactosamine, beta-D-galactose, beta-N-acetyl-D-glucosamine and sialic acid). Additionally, O-acetylated sialic acids were detectable in the glandular acini. The results obtained are discussed with regard to the specific functions of the glandular secretion, which may particularly improve water retention on the skin surface and protect against physical damage as well as microbial contamination. Furthermore, our results support the view of a salivary nature of this gland type.

A case of anaplastic meningioma in a dog.

A tumor sized in 2.0x2.0x2.5 cm developed in the cerebellum of a female Beagle was pathologically investigated. Histopathologically, the tumor grew by compression and partially by infiltration into the adjacent cerebellar parenchyma. There were a large number of necrotic lesions and proliferation of collagen fibers. The tumor cells had oval nucleus showing cellular atypia and a high mitotic index. The tumor cells were reacted with vimentin antibody on immunostain. Electron microscopic examination revealed the tumor cells interdigitated with cytoplasmic processus where the desmosomes developed on cell junction. This tumor was diagnosed as anaplastic meningioma, which is rarely observed in dogs.

Aspects of glycoconjugate production and lysozyme- and defensins-expression of the ceruminous glands of the horse (Equus przewalskii f. dom.).

The distribution of complex glycoconjugates and antimicrobial substances in the ceruminous glands of the horse (Equus przewalskii f. dom., type: pony) was studied using carbohydrate histochemical and immunohistochemical methods. The epithelial cells and luminal secretion of these glands exhibited considerable amounts of glycoconjugates with various saccharide residues, such as alpha-D-mannose, alpha-L-fucose, beta-D-galactose, beta-N-acetyl-D-glucosamine and sialic acid, including O-acetylated sialic acid. Several sugars (alpha-D-mannose, alpha-L-fucose, and beta-D-galactose) were also detectable in the secretion of sebaceous glands present. Additionally, lysozyme and the peptide group of beta-defensins are demonstrated as products of the apocrine ceruminous glands and sebaceous glands. The results obtained are discussed with regard to the functional significance of the glandular secretions. It is suggested that the complex carbohydrates, lysozyme and beta-defensins found in the ceruminous gland secretions are involved in the function of cerumen as a general antimicrobial protective agent in the external auditory canal.

Expression of Secretogranin III in Chicken Endocrine Cells: Its Relevance to the Secretory Granule Properties of Peptide Prohormone Processing and Bioactive Amine Content.

The expression of secretogranin III (SgIII) in chicken endocrine cells has not been investigated. There is limited data available for the immunohistochemical localization of SgIII in the brain, pituitary, and pancreatic islets of humans and rodents. In the present study, we used immunoblotting to reveal the similarities between the expression patterns of SgIII in the common endocrine glands of chickens and rats. The protein-protein interactions between SgIII and chromogranin A (CgA) mediate the sorting of CgA/prohormone core aggregates to the secretory granule membrane. We examined these interactions using co-immunoprecipitation in chicken endocrine tissues. Using immunohistochemistry, we also examined the expression of SgIII in a wide range of chicken endocrine glands and gastrointestinal endocrine cells (GECs). SgIII was expressed in the pituitary, pineal, adrenal (medullary parts), parathyroid, and ultimobranchial glands, but not in the thyroid gland. It was also expressed in GECs of the stomach (proventriculus and gizzard), small and large intestines, and pancreatic islet cells. These SgIII-expressing cells co-expressed serotonin, somatostatin, gastric inhibitory polypeptide, glucagon-like peptide-1, glucagon, or insulin. These results suggest that SgIII is expressed in the endocrine cells that secrete peptide hormones, which mature via the intragranular enzymatic processing of prohormones and physiologically active amines in chickens.

Histochemistry of complex carbohydrates in the ceruminous glands of the goat.

The localization and chemical nature of complex carbohydrates in the ceruminous glands of the Japanese miniature (Shiba) goat were studied using light and electron microscopic histochemical methods, particularly lectin histochemistry. The epithelial cells and luminal secretion of the caprine ceruminous glands contained large amounts of neutral and smaller amounts of acidic glycoconjugates with different terminal sugars (alpha- d-mannose, alpha-L-fucose, alpha-N-acetyl-D-galactosamine, beta-D-galactose, beta-N-acetyl-D-glucosamine, and N-acetyl-neuraminic acid). Several sugars (alpha-L-fucose, beta-D-galactose, beta-N-acetyl-D-glucosamine, and N-acetyl-neuraminic acid) were also detectable in the secretion of the sebaceous glands. The results obtained are discussed with regard to the specific function of the glandular secretion mixture. The complex glycoconjugates found in the ceruminous gland secretion may control viscoelasticity of and bacterial proliferation within the cerumen in order to protect the external auditory canal against physical damage or microbial attacks.

Histochemical analysis of glycoconjugates in the ceruminous glands of the North American raccoon (Procyon lotor).

The distribution and selectivities of glycoconjugates in the ceruminous glands of the North American raccoon (Procyon lotor) were studied by light and electron microscopic histochemical methods, particularly lectin histochemistry. In the modified apocrine glands present, the apocrine secretion mode was combined with exocytosis, whereby the secretory epithelium and the luminal secretion of the ceruminous glands exhibited considerable amounts of complex carbohydrates with various terminal sugars (alpha-D-mannose, beta-D-galactose, alpha-L-fucose, alpha-N-acetyl-galactosamine, beta-N-acetyl-D-glucosamine, N-acetyl-neuraminic acid). Alpha-L-fucose and N-acetyl-neuraminic acid were distinctly prominent in secretory granules or within the free surface coat of the plasma membrane of the glandular cells, as well as in the luminal secretion. Several free sugars (alpha-D-mannose, alpha-L-fucose, beta-D-galactose, beta-N-acetyl-D-glucosamine) were also detectable in the secretion of associated sebaceous glands. The ceruminous gland secretion may control viscoelasticity and/or bacterial degradation of the glandular secretion mixture to improve the protection of the external auditory canal against physical damage or microbial contamination.

Ultracytochemical demonstration of glycoproteins in the canine knee synovium.

By various ultracytochemical methods, glycoconjugates of the synoviocytes, the intercellular matrix and the wall of the small capillaries were studied in the synovial intimal tissues of the canine knee joint. Glycoconjugates with vicinal diol groups could be visualized in certain elements of the Golgi complex, lysosomes, vacuoles, the majority of intracellular cytomembranes, the surface coat of the plasma membrane and glycogen particles in type A cells. In type B cells, less-developed Golgi complexes, and fewer lysosomes and vacuoles were present in the cytoplasm than in that of type A cells. In contrast, a large number of cytoplasmic glycogen particles and abundant vicinal diol-containing groups in the surface coat of the plasma membrane became especially obvious in the B cells. Abundant neutral and acidic glycoproteins were observed in fibrous components in the intercellular matrix. In the small capillaries, strongly positive staining intensities for neutral and acidic glycoconjugates were observed in the basement membrane and perivascular connective tissue, as well as in the surface coat of the luminal plasma membrane of the endothelial cells, although to a somewhat weaker degree. Sialic acid, particularly, was notable in the surface coat of the latter cells. In addition, glycoproteins in the type A cells were shown by lectin ultracytochemistry to contain a variety of saccharide residues such as alpha-D-mannose, alpha-D-glucose, alpha-L-fucose, N-acetyl-beta-D-glucosamine, and N-acetyl-neuraminic acid, which were also found in the plasma membrane of the B cells. The properties of the glycoconjugates found are discussed in relation to the basic functions assigned to the synovial membrane of the canine knee joint.

Ultrastructural localization of hyaluronic acid in the synovium of the goat knee joint.

In the Japanese miniature (Shiba) goat, the synovial membrane contains synoviocytes referred to as type A (macrophage-like cells) and type B cells (fibroblast-like cells) in the intimal layer. Small capillaries and blood vessels of varying sizes were located in the extracellular matrix in the synovial subintima. The type A cells in the synovium possessed numerous vesicles, vacuoles and lysosomes as well as pinocytotic vesicles. These ultrastructural features indicating phagocytosis showed distinct positive reactions following hyaluronan staining. On the other hand, in the type B cells, hyaluronic acids were present in the surface coat of the plasma membrane and its periphery. Additionally, perivascular connective tissue of the small capillaries and blood vessels and interfibrous matrix contained hyaluronan. The results suggest that hyaluronic acid, in the synovial tissue, is synthesized on the plasma membrane of type B cells, and taken up by type A cells. Moreover, hyaluronan is involved in cellular functions in the synovial connective tissue.

Histochemistry of glycoconjugates in the nasolabial skin of the Japanese serow (Capricornis crispus, Artiodactyla, Mammalia) with special reference to glandular structures.

The histochemistry of glycoconjugates in the nasolabial skin of the Japanese serow ( Capricornis crispus ) was studied by light microscopic histochemical methods, particularly lectin histochemistry. The eccrine glands present exhibited neutral and acidic glycoconjugates with different saccharide residues (alpha-L-fucose, beta-D-galactose, beta-N-acetyl-D-glucosamine, alpha-D-galactose and N-acetyl-neuraminic acid) especially in the cells of the secretory acini, the free surface of the collecting duct cells also showed distinct positive reactions with most of the histochemical methods. The thick epidermis of the nasolabial skin contained smaller amounts of glycoproteins. The results obtained are discussed with regard to possible functions of the glandular secretions. This substance mixture may particularly improve water retention on the skin surface, and protect against physical damage as well as microbial contamination. There seem to be no basic differences of muzzle functions between wild and domesticated bovine species.

Cytochemistry of sialoglycoconjugates and lysozyme in canine anal glands as studied by electron microscopic methods.

The distribution of sialoglycoconjugates and lysozyme in the secretory cells of canine anal glands was studied by means of electron microscopic cytochemical methods, particularly lectin cytochemistry and immunocytochemistry. Sialic acids were predominantly present in the secretory granules, Golgi bodies, surface coat of the plasma membrane and luminal secretions. In addition, within these structures, the secretory granules, Golgi bodies and luminal secretions exhibited high levels of sialoglycoconjugates that terminated in Siaα2-6Gal/GalNAc or Siaα2-3Galß1-4GlcNAc. In the secretory cells, reactive gold particles representing lysozyme were mainly detectable in the secretory granules and Golgi bodies. Sialic acids possess diverging functional properties, whereas lysozyme contributes to the non-specific defense against microorganisms. Therefore, their presence and secretion are suggestive of protective effects of both secretory products at the anal mucosa.

Immunohistochemical analysis of IA-2 family of protein tyrosine phosphatases in rat gastrointestinal endocrine cells.

Islet-associated protein-2 (IA-2) and IA-2ß (also known as phogrin) are unique neuroendocrine-specific protein tyrosine phosphatases (PTPs). The IA-2 family of PTPs was originally identified from insulinoma cells and discovered to be major autoantigens in type 1 diabetes. Despite its expression in the neural and canonical endocrine tissues, data on expression of the IA-2 family of PTPs in gastrointestinal endocrine cells (GECs) are limited. Therefore, we immunohistochemically investigated the expression of the IA-2 family of PTPs in the rat gastrointestinal tract. In the stomach, IA-2 and IA-2ß were expressed in GECs that secrete serotonin, somatostatin, and cholecystokinin/gastrin-1. In addition to these hormones, secretin, gastric inhibitory polypeptide (also known as the glucose-dependent insulinotropic peptide), glucagon-like peptide-1, and glucagon, but not ghrelin were coexpressed with IA-2 or IA-2ß in duodenal GECs. Pancreatic islet cells that secrete gut hormones expressed the IA-2 family of PTPs. The expression patterns of IA-2 and IA-2ß were comparable. These results reveal that the IA-2 family of PTPs is expressed in a cell type-specific manner in rat GECs. The extensive expression of the IA-2 family of PTPs in pancreo-gastrointestinal endocrine cells and in the enteric plexus suggests their systemic contribution to nutritional control through a neuroendocrine signaling network.

Cytochemistry of sialoglycoconjugates, lysozyme, and ß-defensin in eccrine glands of porcine snout skin as studied by electron microscopy.

In most mammals except for humanoid primates, eccrine glands are confined to the skin of a series of specific body regions. Sialic acids and antimicrobial substances exhibit various functional properties and serve as a component of nonspecific defense against micro-organisms, respectively. In this study, the distribution of these moieties was studied by electron microscopic histochemical methods. The eccrine glandular acini consisted of two types of dark cells as well as clear cells. The secretory granules and Golgi apparatus of both types of dark cells contained sialic acid residues linked to α2-6Gal/GalNAc. On the other hand, sialoglycoconjugates with Siα2-3Galß1-4GlcNAc sequence were confined to those of the Type II dark cells. In addition, lysozyme and ß-defensin were mainly detected in the secretory granules of the Type II dark cells. These secretory products may create a defensive barrier against microbial invasion and play an essential role in preservation of the integrity of porcine snout skin as a sensory organ.

Histochemical analyses of glycoconjugates and antimicrobial substances in goat labial glands.

Saliva is known to protect the oral cavity and contains glycoproteins and antimicrobial substances. The distribution of these salivary secretions was studied in the labial glands of the Japanese miniature (Shiba) goat using lectin histochemical and immunohistochemical methods. The mucous acinar cells of the labial glands exhibited glycoconjugates with different saccharide residues, such as GalNAcα1-3GalNAc, Galß1-4GalNAc, ß-D-GlcNAc and sialic acid linked to α2-6Gal/GalNAc. Furthermore, α-D-Man, α-L-Fuc, α-D-GalNAc, ß-D-Gal and sialic acid residues were present, in particular, in the serous demilunar cells. Antimicrobial substances (lysozyme, IgA, lactoferrin and ß-defensin) were shown to be mainly immunolocalized in the serous demilunes and duct cells. The results obtained are discussed with regard to the functional role of labial glands. The secretory compounds demonstrated may play an important role in the maintenance of oral health with regard to saliva.