Ultrastructure of feline mammary hypertrophy

The ultrastructure of feline mammary hypertrophy was studied in a five-month-old female which had aborted recently, a ten-year-old female which was one month postestrus, and a four-year-old progestin-treated neutered male. Morphologic comparisons were made to normal mammary tissue from a one-year-old female cat. Hypertrophied mammary tissue had the same cell types and spatial relationships as did the normal gland. Major differences included a more highly developed duct system composed of metabolically active cells which often were arranged in multiple cell layers, and periductular stroma with increased fibroblasts and vascularization. Hypertrophied epithelial cells were characterized generally by smooth-contoured nuclear membranes, more evenly dispersed heterochromatin, prominent nucleoli, increased polyribosomes, and elongated mitochondria. Secretory activity was developed significantly only in the cat that had aborted recently. Modifications in myoepithelial cells included: more evenly dispersed nuclear heterochromatin, thicker bundles of cytoplasmic filaments, more straight plasma membranes along the basal lamina, and elongated hemidesmosomes. Multilayering of the basal lamina was accentuated. Stromal fibroblasts had nuclear heterochromatin distributed similarly to that of epithelial and myoepithelial cells, and increased rough endoplasmic reticulum. Myoepithelial cells did not contribute to the increased stromal cellularity. No significant ultrastructural differences were noted between mammary hypertrophy in young, old, and progestin-treated cats.     

A pleomorphic adenoma of the lacrimal gland in a dog

A 13-year-old female mongrel dog had a pleomorphic adenoma of the lacrimal gland in the right upper orbit. The tumor measured 3.8 x 3.0 x 3.3 cm, appeared white, round, and firm, and pressed the right globe and surrounding tissues. Histopathologically, the tumor had a thin connective tissue capsule and was composed of tubules with two cell types, some resembling luminal epithelial cells making up the tubular structures and the other of myoepithelial cells. Epithelial tubules were disposed in an adenomatous fashion and separated from each other by proliferating pleomorphic myoepithelial cells. Immunohistochemically, large numbers of the luminal epithelial cells revealed an immunopositive reaction against keratin/cytokeratin (AE1/AE3), and some epithelial cells reacted against cytokeratin 14. Spindle-shaped myoepithelial cells revealed an immunopositive reaction against cytokeratin 14, alpha-smooth muscle actin, and vimentin. A small number of myoepithelial cells reacted against desmin. S-100 protein immunopositivity was frequently found in luminal epithelial cells and rarely in the pleomorphic myoepithelial cells. Glial fibrillary acidic protein positivity was commonly found in myoepithelial cells, myxoid matrices, and intracystic materials, but not in luminal epithelial cells.     

The expression of p63 and cytokeratin 5 in mixed tumors of the canine mammary gland provides new insights into the histogenesis of these neoplasms

Cytokeratin 5 and p63 have been described as basal and myoepithelial cell markers in human breast. Mixed tumors of the canine mammary gland have been associated with a myoepithelial origin. Cytokeratin 5 expression has not been evaluated in these tumors. We investigated the relation between cytokeratin 5 and p63 double-immunohistochemical expression in 23 mixed tumors of the canine mammary gland (10 benign mixed tumors and 13 carcinomas arising from benign mixed tumors) and their origin. Cytokeratin 5 and p63 co-expression was observed in myoepithelial cells of benign mixed tumors, as well as in squamous differentiation of carcinoma arising from benign mixed tumors. Though a few interstitial spindle cells of the mesenchymal components expressed both p63 and cytokeratin 5, the basal epithelial cells were labeled only by cytokeratin 5. The co-expression of p63 and cytokeratin 5 in myoepithelial cells and squamous differentiation suggest that, like in human breast, cytokeratin 5 can also be considered a myoepithelial- and squamous-cell differentiating marker in canine tumors. The presence of some interstitial spindle cells stained for p63 and cytokeratin 5 might be associated with a myoepithelial origin of the mesenchymal component of mixed tumors of the canine mammary gland. Moreover, contrary to p63, basal epithelial cells were labeled by cytokeratin 5, indicating that cytokeratin 5 may not represent an exclusive myoepithelial cell marker but also a basal epithelial cell marker in canine mixed tumors. According to these data, basal epithelial cells may be related to the origin of the epithelial component of mixed tumors of the canine mammary gland.     

Alkaline phosphatase reaction of canine mammary mixed tumours: a light and electron microscopic study

Alkaline phosphatase (ALK-P) reactions at the light and electron microscopic levels were performed on eight canine mammary mixed tumours. In the morphologically normal gland next to the tumours, the myoepithelial cells and the stromal tissues near the acinar basement membranes both showed a positive ALK-P reaction by light microscopy. At the electron microscopic level, those parts of the myoepithelial cell membrane that were in contact with other cells--myoepithelial or lumenal epithelial--showed an ALK-P-positive reaction, as did the adjacent stromal tissue. The characteristic tumour cells, at sites of early proliferation, were ALK-P-positive, while in the mucoid and chondroid areas of the mixed tumours they were largely ALK-P-negative by light microscopy. By electron microscopy, those neoplastic cells which were in contact with other cells typically showed a positive ALK-P reaction, while those cells which were in contact with mucoid or chondroid matrix were largely negative. Although the cytoplasmic filaments of the neoplastic cells were 10 nm thick, and those of normal myoepithelial cells were 6 to 8 nm thick, the observations reported provide further evidence for the view that the essential cells of the canine mammary mixed tumour are derived from myoepithelial cells.     

Complex apocrine carcinoma with dominant myoepithelial proliferation in a dog

A rare case of complex apocrine carcinoma displaying dominant myoepithelial proliferation developed in the right leg subcutis of a 10-year-old male dog. The major cell population consisted of diffusely proliferating p63-expressing neoplastic cells that were largely myoepithelial in origin co-expressing α-smooth muscle actin. A small portion of the cell population consisted of concomitant basal epithelial cells lacking α-smooth muscle actin expression. The minor population consisted of p63-negative apocrine gland cells that expressed cytokeratin 8. The myoepithelial cell population showed a rather stronger proliferation activity than did the apocrine epithelial population. Thus, this tumor might have been derived from basal epithelial cells characterized by more predominant myoepithelial differentiation than luminal apocrine epithelial differentiation.     

A Spontaneous Epithelial-Myoepithelial Carcinoma of the Submandibular Gland in a Sprague-Dawley Rat

The present report describes a rare case of spontaneous tumor of the salivary gland in a male Sprague-Dawley rat. The clinically confirmed mass rapidly developed in the cervical region between 19 and 21 weeks of age, and the animal was subsequently euthanized. At necropsy, a well-circumscribed nodule approximately 7 × 6 cm in diameter was found at the site of the salivary gland. The cut surface of the nodule was lobulated and soft and had a pinkish tan fish-flesh appearance. One large cyst (approximately 3 × 2 cm in size) containing reddish fluid was also present in the nodule. Histopathologically, the tumor, with a partially lobulated structure, was surrounded by a thin fibrous capsule. The majority of tumor cells formed a diffuse solid sheet structure that mainly consisted of small ovoid or spindle-shaped cells. In the tumor periphery, some cells were arranged in nest-like structures. Small duct-like structures lined with a monolayer of cuboidal epithelial cells resembling an intercalated duct or large polygonal clear cells with a myoepithelial component were also observed. Mitotic figures and necrotic foci were frequently observed in solid areas. Immunohistochemically, the tumor cells were positive for cytokeratin, epithelial membrane antigen, vimentin, p63, α-smooth muscle actin and calponin. The cells were negative for calcitonin, synaptophysin and chromogranin A. On the basis of these findings, the tumor was diagnosed as an epithelial-myoepithelial carcinoma originating from the luminal epithelial cells and myoepithelial cells in the submandibular gland.     

Immunolocalization of the smooth muscle-specific protein calponin in complex and mixed tumors of the mammary gland of the dog: assessment of the morphogenetic role of the myoepithelium

The immunohistochemical expression of the smooth muscle-specific protein calponin was studied to assess the contribution of myoepithelial cells to the histogenesis of spindle cells of complex and mixed tumors of the mammary gland of the dog and the origin of cartilage and bone in mixed tumors. Formalin-fixed tissues from 55 benign and malignant tumors (49 also containing surrounding normal mammary gland) were evaluated. Periacinar and periductal myoepithelial cells of all the 49 normal mammary glands were diffusely stained by the anti-human calponin monoclonal antibody. Calponin was found in 53 (98%) of the tumors studied, reacting with the myoepithelium-like cells of 86% of benign tumors and their remnants in 85% of malignant tumors. Five different types of calponin-immunoreactive myoepithelial cells were identified: hypertrophic myoepithelial cells. fusiform cells, stellate myoepithelial cells, rounded (myoepithelial) cells, and chondroblasts. Differences in staining intensity and staining pattern among these five types of cells suggested a transition of myoepithelial cells to chondroblasts. Stromal myofibroblasts also showed calponin immunoreactivity, but they did not react with a cytokeratin 14 monoclonal antibody, which recognizes myoepithelial cells in mammary gland. Calponin appears to be a very sensitive marker of normal and neoplastic myoepithelium in the canine mammary gland, and its identification in different cell types of complex and mixed tumors of the mammary gland of the dog suggests a major histogenetic role for myoepithelial cells.     

Alpha basic crystallin expression in canine mammary tumors

The aim of this study was to evaluate prognostic and/or diagnostic factors of canine mammary tumors by immunohistochemically analyzing the expression of alpha basic crystallin (αB-c). For this, formalin-fixed, paraffin-embedded blocks of 51 naturally-occurring canine mammary tumors (11 benign and 40 malignant) were used. Tissue from eight normal canine mammary glands were served as a control. Immunohistochemically, in the control mammary tissues, a few luminal epithelial cells were αB-c positive but myoepithelial cells were negative. In benign or simple type malignant tumors, αB-c expression was observed in luminal epithelial cells while the myoepithelial basal cells were negative. In benign or complex type malign tumors, positive staining was predominantly found in the cytoplasm of epithelial cells. Immunoreactivity of αB-c was also observed in neoplastic myoepithelial cells. Statistically, the number of cells immunolabeled with αB-c was found to be significantly different among tissues from normal canine mammary glands, benign lesions, and malignant tumors (p < 0.05). αB-c immunoreactivity was higher in malignant tumors than the control mammary tissues (p < 0.001). Data obtained in the current study revealed a strong association between high expression levels of αB-c and primary mammary gland tumors in canines.     

Expression of p63 normal canine skin and primary cutaneous glandular carcinomas

p63, a recently identified homologue of the p53 protein, is expressed consistently in basal cells of several human multilayered epithelia. In this study, expression of p63 was determined in 31 primary cutaneous glandular carcinomas, including sebaceous, perianal (hepatoid) gland, apocrine and ceruminous carcinomas, as well as their adjacent normal skin. Similar to humans, p63 is a reliable marker for basal and myoepithelial cells in canine epidermis, cutaneous appendages and malignant apocrine and ceruminous gland neoplasms. In sebaceous carcinomas, not only basal cells, but also some sebocytes, showed nuclear staining for p63. Most mature epithelial cells in perianal gland carcinomas exhibited strong p63 expression. Based on these findings, basal/myoepithelial cells could be involved in the oncogenesis of these tumours and p63 might be used as a diagnostic marker in these lesions.     

Licht- und elektronenmikroskopische Untersuchungen der Samenblasendrüse (Glandula vesicularis) des Bullen

Light and electron microscopical study of the vesicular gland of the bull
The vesicular glands of six adult bulls were studied grossly and with the microscope. Corrosion casts (of the duct system) examined with the dissecting microscope and the SEM clearly showed that the gland was tubulo-alveolar.
The light microscope showed two rows of tall glandular cells. The basal epithelial cells (termed "dwarf lipid-cells") contained uni- as well as plurivacuolar lipid droplets. In the tall glandular cells fine lipid droplets were seen mainly in the supranuclear cytoplasm.
Electron microscopical examinations permit division of the tall glandular cells into three compartments, shown in a semischematic drawing. The results of this study allow the conclusion that the basal "fat cells" and the peroxysomes of the tall glandular cells are related to the formation of prostaglandins.     

Expression of maspin in mammary gland tumors of the dog

Maspin is a serine protease inhibitor that inhibits tumor invasion and metastasis in human breast cancer and is consistently expressed by mammary myoepithelial cells (MECs). To analyze the value of maspin as a marker of the MEC layer of the normal and tumoral canine mammary gland, the immunohistochemical expression of maspin was studied in formalin-fixed tissues from 55 benign and malignant tumors (40 tumors also contained the surrounding normal mammary gland) using a commercially available monoclonal antibody. Periacinar and periductal MECs of all 40 normal mammary glands were stained by the anti-human maspin monoclonal antibody, and immunoreactivity was observed in the nucleus and cytoplasm of these cells. In addition, maspin was found in 53 (98%) of the tumors studied, reacting with the MECs in 100% of benign tumors and 93% of malignant tumors and to the epithelial cells of 16% of benign and 73% of malignant tumors. In the MEC compartment, immunoreactivity was observed in the cytoplasm of hypertrophic MECs, fusiform MECs, stellate MECs, rounded (myoepithelial) cells, and chondroblasts. In the epithelial cell compartment, immunoreactivity was observed in the cytoplasm of cells with and without squamous differentiation. Stromal myofibroblasts were unreactive. Maspin appears to be a very sensitive marker of the normal and neoplastic myoepithelium that, contrary to smooth muscle differentiation markers, does not stain stromal myofibroblasts. In addition, a subset of neoplastic epithelial cells reacted with the maspin antibody. The relationship between maspin expression in different cellular compartments of canine mammary carcinomas and the biologic aggressiveness of the disease remains to be elucidated.     

Distribution of acid and alkaline phosphatases in canine skin.

Azo dye methods were used to determine the distribution of acid phosphatase (ACP) and alkaline phosphatase (ALP) in the skin of 25 Beagle dogs. ALP activity was found in dermal papillae of hair follicles regardless of their state of activity, in Huxley's layer of the inner root sheath of anagen stages, in myoepithelial cells of apocrine sweat glands, in germinative cells of sebaceous glands, in vascular endothelium, and in mast cells. The ACP activity was found in the epidermis, outer and inner root sheaths, keratogenous zone, hair cuticle and medulla, duct of sebaceous gland, and sebum. The results indicate that ACP and ALP are distinctive enzymes serving different biologic functions. The principal role of ALP in the skin appears to be dephosphorylation for adsorption and transport of chemical substances necessary for growth and maintainence of the pilary system and glandular adnexa. The ACP appears to be primarily involved in the breakdown of phospholipids and in necrobiosis of keratinocytes.     

Expression of bone morphogenetic protein-6 (BMP-6) in myoepithelial cells in canine mammary gland tumors.

Seventy-three mammary tumors and three mammary tissue specimens were examined to elucidate the expression of bone morphogenetic protein (BMP)-6 in the myoepithelial cells of canine mammary gland tumors. Morphologically, the myoepithelial cells were classified into four types: resting and proliferating cells inside the basement membrane, and spindle- and star-shaped cells proliferating in the outer area of the basement membrane. The characteristics of these myoepithelial cells were confirmed by immunohistochemistry using antibodies raised against keratin, cytokeratin 19, alpha-smooth muscle actin, and vimentin. In simple adenoma, a small number of resting myoepithelial cells was immunopositive for BMP-6. In complex adenomas and benign mixed tumors, all types of myoepithelial cells, depending in some cases on their specific location within the tumor, were immunopositive for BMP-6, but almost all of the tubular epithelial cells were immunonegative. Foci consisting of a proliferation of BMP-6-positive star- and spindle-shaped cells had mucinous stroma with marked hyaline and chondroid changes. In contrast, the foci with BMP-6-negative spindle- and star-shaped cells tended to have mucinous stroma without chondroid change. Several types of mesenchymal cells including chondrocytes, osteoblasts, and fibroblastlike cells in the mixed tumors, showed an intense immunopositive reaction for the BMP-6 antibody, and were located close to the ectopic cartilage and bone matrix. No significant immunoreactivity for BMP-6 was observed in most of the malignant mammary tumors; only one malignant mixed tumor was examined. All of these findings indicate that BMP-6 expression in myoepithelial cells may increase in complex adenomas and benign mixed tumors in canine mammary glands, and that BMP-6 expression is most intense in the vicinity of chondroid matrix in these tumors.     

In vitro model of parathyroid hormone-related protein secretion from mammary cells isolated from lactating cows

Parathyroid hormone-related protein (PTHrP) is produced by the lactating mammary gland and is present in milk in a biologically active form. The goal of this investigation was to determine if cells cultured from the lactating mammary glands of cows would secrete PTHrP in vitro. Mammary acini were isolated from lactating cows at 1–6 wk after calving, and fresh or cryopreserved mammary acini were cultured for 14 d on Type I collagen. Cultures on thick layers of collagen (2.5 mm) were detached and allowed to contract on Day 6. PTHrP production was measured by N-terminal radioimmunoassay and bioassay (increased cAMP levels in ROS 17/2.8 osteoblast-like cells). The mammary cells reached confluence at Day 6. PTHrP production was low at Day 2 (<0.5 ng/ml) but increased to peak production (2–4 ng/ml) at approximately Day 6 and remained constant until Day 14. Immunoreactive and bioactive PTHrP levels in the culture medium correlated well. The cultures produced lactoferrin (2,000–2,300 ng/ml) and α_s1-casein (14–19 ng/ml). Prolactin stimulated PTHrP production approximately 50% on Days 6–14. PTHrP production was increased approximately 100% by treatment with epidermal growth factor (10 ng/ml) for 2 d. Morphologic evaluation of cultures on thick, contracted collagen at Day 14 revealed an inner layer of mammary epithelial cells overlying myoepithelial cells and an outer layer of collagen containing stromal cells. Immunohistochemistry demonstrated positive staining for PTHrP and cytokeratin in both mammary epithelial and myoepithelial cells and a-smooth muscle actin in myoepithelial cells. These data demonstrated that cryopreserved mammary tissue from lactating cows could be cultured in vitro and secreted PTHrP in a regulated manner. This in vitro model will be useful to investigate the function and regulation of PTHrP in the lactating mammary gland.     

Viral expression and leukocyte adhesion after in vitro infection of goat mammary gland cells with caprine arthritis-encephalitis virus

A characteristic lesion in goats infected by the lentivirus CAEV is mastitis with lymphoid hyperplasia. In order to investigate the mechanism of lesion formation, cultures highly enriched in microvascular endothelial cells, mature and immature luminal epithelial cells, fibroblasts and myoepithelial cells were established from goat mammary gland biopsies. Their susceptibility to in vitro infection with two distinct types of CAEV was investigated by PCR, antigen expression and cytopathy. The capacity of infected mammary gland cells to bind uninfected caprine leukocytes was determined by flow cytometry. All cell types tested were susceptible to CAEV infection in vitro, with different levels of sensitivity according to cell phenotype. Our results suggest that the limited extent of natural infection of mammary gland cells reflects a protective local immune response, and that the myoepithelial cell could act as a reservoir cell. After infection, the mature luminal cell acquires the capacity to bind leukocytes in vitro, which could indicate a facilitation of cellular interactions. The distinct reactions of the different cell types to CAEV infection may be correlated with events leading to progressive lesion development during the natural infection.     

长爪沙鼠颌下腺免疫球蛋白A的定位分布

采用免疫组织化学方法对不同日龄的长爪沙鼠颌下腺IgA的定位分布进行了研究。结果表明，长爪沙鼠的颌下腺由导管部和分泌部构成，分泌部主要由浆液腺构成，导管部包括闰管、纹管、颗粒曲管和小叶间导管等。DAB显色结果表明，IgA阳性细胞主要分布于浆液性腺泡、闰管、纹管、颗粒曲管和小叶间导管，并可分布于腺泡和腺管间结缔组织，IgA阳性产物的分布具有不均一性，无明显随年龄变化的规律性。阳性产物分布于胞质中，胞核呈阴性，对照组阴性。提示从浆细胞产生或循环而来的IgA先经结缔组织进入颌下腺组织，进而定位分布于浆液腺泡和各级导管，导管部有较多的IgA分布。     

DNA measurement and immunohistochemical characterization of epithelial and mesenchymal cells in canine mixed mammary tumours: putative evidence for a common histogenesis.

DNA measurement by image cytometry, and a detailed immunohistochemical study using monoclonal antibodies directed against different human cytokeratin types, muscle-specific actin, vimentin and S100 protein were carried out on normal canine mammary tissue (n =4), benign canine mammary mixed tumours (n =20) and malignant canine mammary mixed tumours (n =13). The results showed that ductal and alveolar luminal cells in normal and neoplastic tissue were immunoreactive with CAM5.2 and AE1/AE3 antibodies recognizing human keratins.Basal/myoepithelial cells were clearly differentiated from ductal and alveolar epithelial cells, since the latter only expressed cytokeratins, whereas the former also expressed vimentin and muscle-specific actin. This immunohistochemical study showed that there is loss of expression of muscle-specific actin and cytokeratins in areas of myoepithelial proliferation, and enhanced expression of vimentin and S100 protein in proliferative areas with osseous and/or chondroid metaplasia. The ploidy studies revealed that 20% (4/20) of benign and 54% (7/13) of malignant mixed tumours of canine mammary gland were aneuploid and that the epithelial and myoepithelial components of the mixed tumours had identical DNA content.Our results reinforce the role of myoepithelial cells in mesenchymal metaplasia in mixed mammary tumours and suggest the possibility of a common origin of both components from a totipotential stem cell with capacity for divergent differentiation.     

The fine structure of the Harderian gland in the ostrich (Struthio camelus)

The Harderian gland of the ostrich (Struthio camelus) is a tubuloalveolar gland containing holocrine secreting epithelial cells. The gland epithelium is composed of two different cell types, which can be classified as type I and type II. These cells contain dense secretory vesicles in their cytoplasm and they are connected laterally with desmosomes. At the basal site of these cells, myoepithelial cells are present. Plasma cells are observed in the subepithelial region of the gland. In the interlobular trabeculae, forming the gland stroma, fibroblasts, blood vessels and nerve fibres are included. Another important finding in the ostrich Harderian gland is the presence of homogeneous material.     

Histological variations in myoepithelial cells and arrectores pilorum muscles among caudal, metatarsal and preorbital glands in Hokkaido sika deer (Cervus nippon yesoensis Heude, 1884)

The morphological characteristics of myoepithelial cells and arrectores pilorum muscles were investigated in caudal, metatarsal and preorbital glands of Hokkaido sika deer (Cervus nippon yesoensis Heude, 1884) using immunohistochemistry for alpha-smooth muscle actin. In the metatarsal, preorbital and general skin glands, myoepithelial cell layers continuously embraced the secretory epithelium, while in the caudal gland, discontinuous myoepithelial cell rows surrounded the apocrine tubules. There was a trend that the widths of the myoepithelial cells of the caudal and preorbital glands appeared to be thinner than those of the metatarsal and general skin glands. In the metatarsal gland, the arrectores pilorum muscles were highly developed and considerably larger than those in other skin glands.     

乳腺中脂肪细胞的转化和更迭

雌性动物的乳腺组织是由胚胎时期的外胚层发育而来,出生时只有少量导管和皮下基质结构,而后在性成熟、妊娠和泌乳期乳腺发育达到峰值,这一特点使乳腺成为出生后唯一可以重复再生的器官。在乳腺发育到退化的循环中,乳腺的上皮细胞、基质白色脂肪细胞、棕色脂肪细胞和肌上皮细胞经历了一系列转化和更迭,脂肪细胞的动态转化反映了乳腺的功能变化。研究乳腺细胞的转化和更迭与母畜的泌乳直接相关,对延续母畜的生产效率有重要意义。本文就乳腺中脂肪细胞转化方面的最新研究进展进行综述,为深度揭示乳腺发育过程中细胞更迭的机制提供前沿研究信息。