Takayuki Nara

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

The present study revealed in detail the subcellular localization of lysozyme and β-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 β-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.

Histochemical properties of sialic acids and antimicrobial substances in canine anal glands

The functional properties of sialic acids appear to be manifold. Additionally, antimicrobial substances serve as a non-specific defense against microorganisms. In this study, therefore, the localization of sialic acids and antimicrobial substances in the anal glands of dog was studied by sialoglycoconjugate histochemistry and immunohistochemistry. The secretory epithelium, luminal secretions and excretory ducts exhibited high levels of sialoglycoconjugates that terminated in Siaα2-6Gal/GalNAc or Siaα2-3Gal1-4GlcNAc. Additionally, O-acetylated sialic acids were detectable in these glandular structures. Antimicrobial substances, such as lysozyme, immunoglobulin A (IgA), lactoferrin and the peptide group of β-defensins, were also demonstrated as products of the anal glands. The results obtained are discussed with regard to the functional significance of the anal glands. These secretory products may create a defensive barrier against microbial invasion at the anal mucosa.

Morphological, histochemical and immunohistochemical characterization of secretory production of the ciliary glands in the porcine eyelid

In addition to performing general histology and cytology of the ciliary glands of the miniature pig, we studied the localization of glycoconjugates and beta-defensins in these glands with the use of carbohydrate histochemical and immunohistochemical methods. The secretory cells of the glands were equipped with non-homogeneous secretory granules, a well-developed Golgi apparatus and rough endoplasmic reticulum. The secretory epithelium and luminal secretion of the glands contained large amounts of neutral and acidic glycoconjugates with various saccharide residues (alpha-L-Fuc, beta-D-Gal, alpha-D-GalNAc and sialic acid). The sebaceous glands and tarsal glands also exhibited positive reactions to most of the histochemical methods. Additionally, the antimicrobial peptide group of beta-defensins was demonstrated to be products of the ciliary glands, as well as the sebaceous glands and tarsal glands. The results obtained are discussed with regard to the specific function of the ciliary glandular secretions. These secretory products may be related to the moistening and general protection of the skin surface of the eyelid and ocular surface.

Functional properties of feline foot pads as studied by lectin histochemical and immunohistochemical methods.

The localization of sialic acids and antimicrobial substances in the foot pads of the cat was examined by lectin histochemical and immunohistochemical methods. The lectin binding patterns of the eccrine glands were suggestive of the existence of large concentrations of sialoglycoconjugates that terminated in Siaalpha2-3Gal1-4GlcNAc. Results were consistent with localization of O-linked (mucin-type) sialoglycoproteins with the Siaalpha2-6Gal/GalNAc sequence in the epidermal layers, especially the stratum spinosum. Additionally, antimicrobial peptides, such as lysozyme, secretory component, lactoferrin, and the peptide group of beta-defensins were demonstrated to be immunolocalised in the eccrine glandular cells. These substances, except for secretory component, were also distributed in the epidermal strata. The sialic acids and antimicrobial substances found in the eccrine glandular secretions and epidermis may play an essential role in the preservation of skin integrity in feline foot pads.

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 (α-d-mannose, α-l-fucose, α-N-acetyl-d-galactosamine, β-d-galactose, β-N-acetyl-d-glucosamine, and N-acetyl-neuraminic acid). Several sugars (α-l-fucose, β-d-galactose, β-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 demonstration of sialic acids and antimicrobial substances in the porcine anal glands.

The distribution of sialic acids and antimicrobial products (lysozyme, β-defensin-1, lactoferrin, IgA) in the anal glands of miniature pig was studied by glycoconjugate histochemistry and immunohistochemistry. The glandular acini of these glands exhibited considerable amounts of sialoglycoconjugates that terminated in Siaα2-6Gal/GalNAc or Siaα2-3Gal1-4GlcNAc, including O-acetylated sialic acids. Additionally, all antimicrobial products examined could be demonstrated in the anal glands, especially in the serous cells. The results obtained are discussed with regard to the functional significance of the anal glands. Our observations corroborated the view that sialic acids closely interact with defense cells and antimicrobial substances in the innate immune response. Therefore, the anal glandular secretions may function as protective agents in order to preserve the integrity of the anal region.

Histochemical Analyses of Anti-Microbial Substances in Canine Perianal Skin with Special Reference to Glandular Structures

Circumanal glands are prominent features of the canine perianal skin, which are often located near to the sebaceous glands and apocrine glands. As the functional relevance of circumanal glands is yet unknown, we studied the localisation of sialic acids and anti‐microbial substances (lysozyme, immunoglobulin A, lactoferrin, β‐defensin) in these glandular structures by lectin histochemistry and immunohistochemistry. The glands exhibited a number of sialic acids that were linked to α2‐6Gal/GalNAc and α2‐3Galβ1‐4GlcNAc. Additionally, lysozyme, lactoferrin and β‐defensin could be demonstrated in the three types of skin glands, whereas IgA was only detectable in the apocrine glands. The results of the study suggest the specific significance of the circumanal glands. Independent of a certain endocrine role, their products may mainly function as protective agents to preserve the integrity of the anal region, considering that sialic acids and anti‐microbial substances are important in defence mechanisms.

Cytochemical Localization of Glycoconjugates in the Secretory Epithelial Cells Lining the Prostate Gland of the Miniature Pig

The aim of the study was to emphasize several specific functional aspects of the mammalian prostate gland. In this connection, the subcellular localization and characterization of glycoconjugates in the secretory epithelial cells were examined that line the prostate gland of the miniature pig, using cytochemistry, including lectin‐gold methods. The results verified a diversified pattern of glycoconjugates in the structures of the secretory cells. The secretory granules and the free surface coat of the plasma membrane contained several sugar moieties such as α‐D‐Man, α‐D‐Glc, α‐L‐Fuc, β‐D‐Gal, GalNAc, GlcNAc and NeuAc (sialic acid), whereas the Golgi apparatus exhibited GlcNAc and sialic acid residues. In addition, the cisterns of the endoplasmic reticulum showed α‐D‐Man and α‐D‐Glc moieties. It seems likely that the secretory glycoconjugates demonstrated contribute to favourable conditions for fertilization.

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 Siaα2-3Galβ1-4GlcNAc. 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.

Sialic acids and antimicrobial substances in the apocrine glands of porcine perianal skin

The porcine perianal skin shows prominent apocrine glands with large saccular dilatations, whereby the functional significance of the glandular secretions is rather unexplained. Our study focuses on the demonstration of sialoglycoconjugates and antimicrobial substances in these glands, using glycoconjugate histochemical and immunohistochemical methods. The result obtained emphasized the general presence of sialic acids, linked to α2-6Gal/GalNAc and α2-3Gaβl1-4GlcNAc, in the secretory cells. The secretory epithelium and luminal secretions also contained a spectrum of antimicrobial substances, such as lysozyme, IgA, lactoferrin, and the peptide group of β-defensins. Realizing that sialic acids possess diverging functional properties through various saccharide residues, and that antimicrobial substances serve as a non-specific defense against microorganisms, these secretory products may function as protective agents in order to preserve the integrity of the perianal region. This view includes that the amounts of bacteria on the skin surface are controlled and maintained at the certain level.