Normal Canine Mammary Glands Research Articles

Growth hormone induces tyrosyl phosphorylation of the transcription factors Stat5a and Stat5b in CMT-U335 canine mammary tumor cells

It has now been well documented that the normal and tumorous canine mammary glands can be extra-pituitary sites of substantial growth hormone (GH) synthesis. Until now, attempts to reproduce the GH synthesis in-vitro using canine mammary explants or mammary tumor cells have not been successful. Therefore, the response of CMT-U335 canine mammary tumor cells to administered porcine GH (pGH) was investigated as an in-vitro model to study the possible effects of GH synthesis on this site. CMT-U335 cells spontaneously express the growth hormone receptor (GHR) as well as the prolactin receptor (PRLR). Twenty five minutes after administration, GH induced, in a dose-dependent manner, phosphorylation of the transcription factors Stat5a and Stat5b. Clear phosphorylation was induced by 10 −7 M and 10 −8 M pGH, with virtually no phosphorylation at 10 −9 M pGH. A similar dose-dependent phosphorylation of Stat5a by ovine prolactin was found in these cells. Although at high concentrations binding of pGH to the canine PRLR can occur (albeit with a low pKa), the similar dose-dependent effect of oPRL on Stat5a phosphorylation indicated that pGH signaled through the GHR. Remarkably, pGH induced a moderately decreased proliferation of CMT-U335 tumor cells, which may indicate that GH induces differentiation in these tumor cells. The GH-induced activation of Stat5a and Stat5b in these cells, as part of the JAK/Stat signal transduction pathway, is consistent with mammary GH playing a role in autocrine and/or paracrine stimulation of (tumorous) mammary cells.

Immunohistochemical study of hormonal receptors and cell proliferation in normal canine mammary glands and spontaneous mammary tumours

The expression of hormone receptors and their relationship_to cell proliferation in six samples of normal canine mammary tissue, and 11 benign and 10 malignant mammary neoplasms from female dogs were...

Evaluation of a commercial system for the identification of Ornithobacterium rhinotracheale.

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Lectin Histochemistry of Normal and Neoplastic Canine Mammary Gland Epithelial Cells.

Lectin histochemical study was done on canine mammary gland epithelial cells of 6 groups (normal, hyperplasia, adenoma, adenocarcinoma, benign mixed tumor and malignant mixed tumor). Among ten lectins used, three lectins showed an apparent alteration in the binding activity following neoplastic changes. Namely, Dolichos biflorus and Helix pomatia specific for terminal N-acetylgalactosamine residue bound more strongly to the surface of epithelial cells in adenocarcinoma and malignant mixed tumor. In addition, Ulex europaeus specific for terminal fucose residue showed stronger binding activity in mammary neoplasma irrespective of malignancy.

Immunohistochemical distribution of type IV collagenase in normal, dysplastic and neoplastic canine mammary gland

The distribution of type IV collagenase (cIVase) was evaluated immunohistochemically in a series of normal, dysplastic and neoplastic canine mammary glands, as well as in lymph node metastases. In normal and dysplastic mammary tissues, and benign mammary tumours, cIVase was observed in myoepithelial cells, but in malignant tumours it was mainly localized in the cytoplasm of carcinoma cells and fibroblasts close to infiltrating neoplastic cells. No significant difference was observed between carcinomas of different histological sub-types. These results suggest that the distribution of cIVase is a potentially useful indicator of malignancy in canine mammary tumours.

E cadherin expression in normal and neoplastic canine mammary gland

The epithelial intercellular adhesion molecule, E cadherin, was evaluated immunohistochemically in a series of normal and malignant neoplastic canine mammary glands. The normal glands showed strong immunolabelling at the intercellular borders of all the ductal cells. Tubular and papillary mammary adenocarcinomas had reduced intercellular E cadherin positivity with "heterogeneous" or "cytoplasmic" distribution. Solid and squamous cell carcinomas were negative. These results suggest that reduced expression and abnormal distribution of E cadherin may be related to the differentiation grade of these neoplasms.

Expression of β1 integrin in normal, dysplastic and neoplastic canine mammary gland

The expression of beta 1 integrin was evaluated immunohistochemically in a series of normal, dysplastic and neoplastic canine mammary glands, and in lymph node metastases. The tissues were formalin-fixed and paraffin wax-embedded. In malignant neoplasms, beta 1 integrin was decreased and redistributed along the entire cell membrane. In lymph nodes, strong immunohistochemical staining was seen intercellularly in clusters of metastatic cells within subcapsular sinuses and at the periphery of intranodal metastases. These results suggest that the expression of integrin molecules may be related to malignancy and to the metastatic potential of neoplastic cells.

Expression of fibronectin and its integrin receptor α5β1 in canine mammary tumours

Fibronectin and its integrin receptor α5β1 were studied by immunohistochemical methods in five normal canine mammary glands, four dysplastic glands and 18 mammary tumours. The aim of the study was to evaluate the possible changes in the α5β1 integrin receptor and its ligand fibronectin in relation to the metastatic capacity of canine mammary neoplasms. The immunostaining of α5β1 was very uniform in the hyperplastic glands but uneven in the mammary tumours. The expression of α5 and β1 was diminished in metastatic tumours but there were some α5-positive cells with pronounced features of malignancy and immaturity. Stromal fibronectin was increased in most cases and cytoplasmic staining of fibronectin was observed in epithelial and myoepithelial cells in mammary neoplasms but not in normal or dysplastic mammary tissue. There was no relationship between the content of α5β1 and the expression of fibronectin in canine mammary tumours.

S100 Protein is Not Specific for Myoepithelial Cells in the Canine Mammary Gland

The distribution of protein S100 in the canine mammary gland was studied, this substance having been described as specific for myoepithelial cells. Two immunohistochemical methods, peroxidase-anti-peroxidase complex (PAP) and alkaline phosphatase-anti-alkaline phosphatase complex (APAAP) were used on frozen sections of normal canine mammary gland. Both the myoepithelial and epithelial cells stained positively with PAP. With APAAP, staining was also seen in the controls, presumably because endogenous alkaline phosphatase was not blocked. As protein S100 was shown to be present both in myoepithelial and epithelial cells of the mammary gland, it was concluded that S100 is not a specific marker for myoepithelial cells.

Immunohistochemistry with keratin, vimentin, desmin, and alpha-smooth muscle actin monoclonal antibodies in canine mammary gland: malignant mammary tumours.

Ten malignant canine mammary gland tumours and five metastases from three of these tumours were studied immunohistochemically with monoclonal antibodies (MoAbs) directed against different human keratin types (K), alpha-smooth muscle actin, vimentin, and desmin. In all tumours the neoplastic epithelium was rather homogeneously labelled with the keratin MoAbs RCK 102 (K 5 and 8) and CAM 5.2 (K 8). The adenocarcinomas (n = 5), the solid carcinomas (n = 2), and the carcinosarcoma (n = 1) showed heterogeneous labelling with the MoAbs specific for luminal cell antigens in the normal canine mammary gland, i.e., K 18, K 7 and K 19 MoAbs. These cells were also immunoreactive with K 4 and K 10 MoAbs. The spindle cell carcinomas (n = 2), however, did not react with these MoAbs. All tumours except one adenocarcinoma were characterized by the absence of immunoreactive labelling with the alpha-smooth muscle actin MoAb. In the solid carcinomas this was associated with the absence of labelling with one or both basal cell specific keratin MoAbs, i.e., 8.7 (K 14 and 17) and RCK 107 (K 14), respectively. In contrast, the other malignant tumours showed marked labelling of neoplastic epithelium with these MoAbs. Another remarkable finding was the labelling of a limited to moderate number of neoplastic epithelial cells with the vimentin MoAb. The presence of such labelling patterns in canine mammary gland tumours may be indicative of malignancy. Metastatic tumour tissues had a labelling pattern largely similar to that of the primary tumour, although also loss of reactivity for some keratin MoAbs was seen.

Immunohistochemistry with keratin, vimentin, desmin, and alpha-smooth muscle actin monoclonal antibodies in canine mammary gland: normal mammary tissue.

Normal canine mammary gland tissue was studied immunohistochemically with monoclonal antibodies (MoAbs) directed against various human keratin types, vimentin, desmin, and alpha-smooth muscle actin. Both ductal and alveolar luminal cells were immunoreactive with MoAbs recognizing respectively human keratins no. 7, 8, 18 and 19. In addition, some ductal luminal cells were labelled with a keratin 4 and a keratin 10 MoAb. Basal/myoepithelial cells were immunoreactive only with MoAbs directed against keratin 14, keratins 14 and 17, and alpha-smooth muscle actin. The vimentin MoAb merely labelled solitary loose intraluminal cells representing macro-phages or sloughed epithelial cells. These findings correspond largely to observations made in human breast tissue.

Immunohistochemical Reactivity of Basal and Luminal Epithelium-specific Cytokeratin Antibodies within Normal and Neoplastic Canine Mammary Glands

Human basal epithelium (myoepithelium)-specific (312C8-1) and luminal epithelium-specific (13H5) cytokeratin antibodies were applied to frozen sections of normal canine mammary tissues (seven), benign adenomas and hyperplasias (five), mixed tumors (12), and adenocarcinomas (18) to determine if epithelial subsets could be discriminated by the use of an avidin biotin peroxidase complex immunohistochemical procedure. The 312C8-1 and 13H5 antibodies were consistently reactive with basal and luminal epithelium, respectively, in the normal mammary gland (7/7) and in benign adenomas and hyperplasias (5/5). Mixed mammary tumors had similar basal and luminal epithelial reactivity and also had proliferating spindle-shaped stromal cells that were reactive with 312C8-1 (10/12) and 13H5 (4/12). The adenocarcinomas were subclassified into basal, luminal, and basal/luminal on the basis of 312C8-1 reactivity (4/18), 13H5 reactivity (2/18), and dual reactivity with mutually exclusive anatomic distribution (11/18), respectively. Those tumors with dual immunoreactivity were indicative of noninvasive carcinomas. Dogs with neoplasms that were reactive with 312C8-1 and nonreactive with 13H5 had local recurrence or distant metastasis within 2 weeks to 6 months after diagnosis. Other antibodies used for comparison were pan cytokeratin AE1/AE3, actin HHF35, and vimentin. 312C8-1 and 13H5 antibodies are specific for canine mammary basal and luminal epithelium, respectively, and by employing these antibodies, the origin and differentiation of canine mammary neoplasms can be determined more accurately than on the basis of hematoxylin and eosin-stained tissue alone.

Glucose-6-phosphate dehydrogenase and 6-Phosphogluconate dehydrogenase activities in normal canine mammary gland and in mammary tumours and their correlation with oestrogen

Glucose-6-phosphate dehydrogenase (G-6-PD) and 6-phosphogluconate dehydrogenase (6-PGD) activities were studied in 10 normal canine mammary glands and in 19 canine mammary tumours, which included seven benign and 12 malignant neoplasms. The malignant tumours were also examined for oestradiol receptors. The mean G-6-PD and 6-PGD activities were 5.17 +/- 1.84 and 1.65 +/- 0.64 IU per g protein in normal glands, 8.8 +/- 2.7 and 3.8 +/- 0.99 IU per g protein in benign and 19.6 +/- 5.2 and 8.5 +/- 2.1 IU per g protein in malignant mammary tumours. The enzyme activities were significantly higher in tumour tissue than in normal glands. Malignant tumours had much greater activity than benign. With the increasing anaplasia, there was a corresponding rise in the activity of the two enzymes. There was an inverse correlation between the oestrogen receptor (ER) status and the enzyme activity. The ER-positive tumours had low and ER-negative tumours had high enzyme activities.

Ontogenetic aspects of nonspecific hyperventilation effect in children and its relation to diagnosis of seizures

Canine mammary tumours (CMTs) are the most common tumours of entire female dogs and represent a promising model for human breast cancer. Little is known about the presence and prognostic value of lymphangiogenesis in CMTs. The aims of the present study were to analyze selected characteristics of lymphatic vessels in CMTs, to evaluate their prognostic significance and to compare these results with studies of human breast cancer. Fifty-six benign CMTs, 55 malignant CMTs and 13 control samples of normal canine mammary gland tissue were studied. Serial immunohistochemical labelling with the lymphatic marker prox-1 and the proliferation marker Ki67 was performed. In intratumoural (IT) and peritumoural (PT) regions, the lymphatic vessel density (LVD), mean lymphatic vessel perimeter (LVP) and relative area occupied by lymphatic vessels (LVA) were analyzed. Lymphatic endothelial cell proliferation (LECP) and tumour cell proliferation (TCP) were also measured. Lymphatic vessels were identified in IT and PT regions and lymphangiogenesis was present in both regions. The IT lymphatic vessels were smaller, less numerous and occupied a smaller relative area compared with those of the PT region. Although no differences in lymphatic vessel parameters were observed between benign and malignant tumours, control tissue differed significantly from neoplastic tissue. None of the lymphatic vessel parameters showed a prognostic value, except for LECP in PT regions of benign tumours. The findings were in accordance with results of investigations into human breast cancer, which supports the use of dogs with spontaneously occurring CMTs as an animal model in comparative oncology trials.