A new monoclonal antibody to a cell-surface antigen that distinguishes luminal epithelial and myoepithelial cells in the rat mammary gland

1989 ◽  
Vol 94 (3) ◽  
pp. 545-552
Author(s):  
R.S. Mahendran ◽  
M.J. O'Hare ◽  
M.G. Ormerod ◽  
P.A. Edwards ◽  
R.A. McIlhinney ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Mammary Gland ◽  
Epithelial Cells ◽  
Milk Fat ◽  
Single Cells ◽  
Myoepithelial Cells ◽  
Stages Of Development ◽  
Leukaemia Cell Line ◽  
Flow Cytometric ◽  
Rat Mammary Gland

A monoclonal antibody (25.5) has been produced that recognises luminal epithelial cells of the rat mammary gland. This antibody together with monoclonal anti-CALLA antibodies, which react with mammary myoepithelial cells, has been used in biochemical, immunocytochemical and flow cytometric studies. Antibody 25.5 bound to proteins of molecular weight 70K and 25K (K = 10(3) Mr) in both the rat milk fat globule membrane and in single cell suspensions prepared from the virgin adult rat mammary gland. Anti-CALLA antibody (J5), recognised a 93–100K protein in the gland extracts, which co-electrophoresed with the CALLA/CD-10 antigen from NALM-6 acute lymphoblastic leukaemia cell line. Antibody 25.5 bound to the luminal surface of rat mammary epithelial cells at all stages of development from neonatal through to pregnancy, lactation and involution. CALLA immunoreactive staining has previously been shown on basally located presumptive myoepithelial cells at all stages of development. Flow cytometric analyses demonstrated that 25.5 and anti-CALLA antibodies stained independent cell populations in suspensions of single cells prepared from purified epithelial elements from the mammary gland of adult virgin rat.

scholarly journals Expression of Maspin in Mammary Gland Tumors of the Dog

2005 ◽  
Vol 42 (3) ◽  
pp. 250-257 ◽  
Author(s):  
A. Espinosa de los Monteros ◽  
M. Y. Millán ◽  
G. A. Ramírez ◽  
J. Ordás ◽  
C. Reymundo ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Mammary Gland ◽  
Epithelial Cells ◽  
Malignant Tumors ◽  
Myoepithelial Cells ◽  
Benign Tumors ◽  
Normal Mammary Gland ◽  
Differentiation Markers ◽  
Maspin Expression ◽  
Cellular Compartments

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.

scholarly journals Distribution of myoepithelial cells and basement membrane proteins in the resting, pregnant, lactating, and involuting rat mammary gland.

1982 ◽  
Vol 30 (7) ◽  
pp. 667-676 ◽  
Author(s):  
M J Warburton ◽  
D Mitchell ◽  
E J Ormerod ◽  
P Rudland
Keyword(s):  
Mammary Gland ◽  
Membrane Proteins ◽  
Epithelial Cells ◽  
Connective Tissue ◽  
Basement Membrane ◽  
Myoepithelial Cells ◽  
Type I ◽  
Continuous Layer ◽  
Basement Membrane Proteins ◽  
Rat Mammary Gland

Using antisera to specific proteins, the localization of the rat mammary parenchymal cells (both epithelial and myoepithelial), the basement membrane, and connective tissue components has been studied during the four physiological stages of the adult rat mammary gland, viz. resting, pregnant, lactating, and involuting glands. Antisera to myosin and prekeratin were used to localize myoepithelial cells, antisera to rat milk fat globule membrane for epithelial cells, antisera to laminin and type IV collagen to delineate the basement membrane and antisera to type I collagen and fibronectin as markers for connective tissue. In the resting, virgin mammary gland, myoepithelial cells appear to form a continuous layer around the epithelial cells and are in turn surrounded by a continuous basement membrane. Antiserum to fibronectin does not delineate the basement membrane in the resting gland. The ductal system is surrounded by connective tissue. Only the basal or myoepithelial cells in the terminal end buds of neonatal animals demonstrate cytoplasmic staining for basement membrane proteins, indicating active synthesis of these proteins during this period. In the secretory alveoli of the lactating rat, the myoepithelial cells no longer appear to form a continuous layer beneath the epithelial cells and in many areas the epithelial cells appear to be in contact with the basement membrane. The basement membrane in the lactating gland is still continuous around the ducts and alveoli. In the lactating gland, fibronectin appears to be located in the basement membrane region in addition to being a component of the stroma. During involution, the alveoli collapse, and appear to be in a state of dissolution. The basement membrane is thicker and is occasionally incomplete, as also are the basket-like myoepithelial structures. Basement membrane components can also be demonstrated throughout the collapsed alveoli.

MicroRNA-432 inhibits milk fat synthesis by targeting SCD and LPL in ovine mammary epithelial cells

2021 ◽  
Author(s):  
Zhiyun Hao ◽  
Yuzhu Luo ◽  
Jiqing Wang ◽  
Jon Hickford ◽  
Huitong Zhou ◽  
...  
Keyword(s):  
Mammary Gland ◽  
Epithelial Cells ◽  
Milk Production ◽  
Milk Fat ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Differentially Expressed ◽  
Differentially Expressed Mirnas ◽  
Fat Synthesis ◽  
Milk Fat Synthesis

In our previous studies, microRNA-432 (miR-432) was found to be one of differentially expressed miRNAs in ovine mammary gland between the two breeds of lactating sheep with different milk production...

scholarly journals Enhanced synthesis of gelatinase and stromelysin by myoepithelial cells during involution of the rat mammary gland.

1992 ◽  
Vol 40 (5) ◽  
pp. 697-703 ◽  
Author(s):  
S R Dickson ◽  
M J Warburton
Keyword(s):  
Mammary Gland ◽  
Basement Membrane ◽  
Gelatin Zymography ◽  
Mammary Glands ◽  
Myoepithelial Cells ◽  
Immunofluorescence Staining ◽  
Rat Mammary Gland

During the involution of the mammary gland there is destruction of the basement membrane as the secretory alveolar structures degenerate. Immunofluorescence staining of sections of rat mammary gland with antibodies to 72 KD gelatinase (MMP-2) and stromelysin (MMP-3) revealed increased production of these two proteinases during involution. This increased expression was mostly restricted to myoepithelial cells. Increased expression during involution was also demonstrated by immunoblotting techniques. Gelatin zymography indicated that the predominant metalloproteinase present in involuting rat mammary glands was a 66 KD gelatinase.

Regeneration of mammary glands in vivo from isolated mammary ducts

Development ◽  
1986 ◽  
Vol 96 (1) ◽  
pp. 229-243
Author(s):  
E. Jane Ormerod ◽  
Philip S. Rudland
Keyword(s):  
Epithelial Cells ◽  
Milk Fat ◽  
Cell Types ◽  
Mammary Glands ◽  
Myoepithelial Cells ◽  
Basal Cells ◽  
Alveolar Cells ◽  
Terminal End Buds ◽  
Mammary Cell

Rat mammary ducts, free of buds, can alone regenerate complete mammary trees when transplanted into the interscapular fat pads of syngeneic host rats. All the main mammary cell types are identified within such outgrowths. Epithelial cells, which show the presence of milk fat globule membrane antigens and microvilli on their luminal surfaces, line the ducts. Basal cells surrounding the ducts show characteristic features of myoepithelial cells: immunoreactive actin and keratin within the cytoplasm, myofilaments, pinocytotic vesicles and hemidesmosomal attachments to the basement membrane. Cells within the end buds and lateral buds, however, show few if any cytoplasmic myofilaments and are relatively undifferentiated in appearance. Intermediate morphologies between these cells and myoepithelial cells are seen nearer the ducts. In this respect they exactly resemble the cap cells found in terminal end buds (TEBs) of normal mammary glands. Occasional epithelial cells within alveolar buds show the presence of immunoreactive casein, which is a product of secretory alveolar cells in the normal rat mammary gland. Dissected terminal end buds can regenerate similar ductal outgrowths. Thus, ductal tissue alone can generate all the major mammary cell types seen in the normal gland, including the cap cells.

Milk fat globule is an alternative to mammary epithelial cells for gene expression analysis in buffalo

2016 ◽  
Vol 83 (2) ◽  
pp. 202-208 ◽  
Author(s):  
Qiuming Chen ◽  
Yanjun Wu ◽  
Mingyuan Zhang ◽  
Wenwen Xu ◽  
Xiaoping Guo ◽  
...  
Keyword(s):  
Gene Expression ◽  
Mammary Gland ◽  
Epithelial Cells ◽  
Expression Analysis ◽  
Milk Fat ◽  
Gene Expression Analysis ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Milk Fat Globule ◽  
Fat Globule

Owing to the difficulty in obtaining mammary gland tissue from lactating animals, it is difficult to test the expression levels of genes in mammary gland. The aim of the current study was to identify if milk fat globule (MFG) in buffalo milk was an alternative to mammary gland (MG) and milk somatic cell (MSC) for gene expression analysis. Six buffalos in late lactation were selected to collect MFG and MSC, and then MG was obtained by surgery. MFG was stained with acridine orange to successfully visualise RNA and several cytoplasmic crescents in MFG. The total RNA in MFG was successfully isolated and the integrity was assessed by agarose gel electrophoresis. We analysed the cellular components in MFG, MG and MSC through testing the expression of cell-specific genes by qRT-PCR. The results showed that adipocyte-specific gene (AdipoQ) and leucocyte-specific genes (CD43, CSF1 and IL1α) in MFG were not detected, whereas epithelial cell marker genes (Keratin 8 and Keratin 18) in MFG were higher than in MSC and lower than in MG, fibroblast marker gene (vimentin) in MFG was significantly lower than in MG and MSC, milk protein genes (LALBA, BLG and CSN2) and milk fat synthesis-related genes (ACC, BTN1A1, FABP3 and FAS) in MFG were higher than in MG and MSC. In conclusion, the total RNA in MFG mainly derives from mammary epithelial cells and can be used to study the functional gene expression of mammary epithelial cells.

scholarly journals Leptin Expression in Human Mammary Epithelial Cells and Breast Milk

1998 ◽  
Vol 83 (5) ◽  
pp. 1810-1810 ◽  
Author(s):  
Susan M. Smith-Kirwin ◽  
Darlise M. O’Connor ◽  
Jennifer Johnston ◽  
Elizabeth de Lancy ◽  
Sandra G. Hassink ◽  
...  
Keyword(s):  
Mammary Gland ◽  
Breast Milk ◽  
Epithelial Cells ◽  
Milk Fat ◽  
Mammary Epithelial Cells ◽  
Skim Milk ◽  
Mammary Epithelial ◽  
Pcr Analysis ◽  
Human Mammary Epithelial Cells ◽  
Human Mammary Epithelial

Leptin has recently been shown to be produced by the human placenta and potentially plays a role in fetal and neonatal growth. Many functions of the placenta are replaced by the mammary gland in terms of providing critical growth factors for the newborn. In this study, we show that leptin is produced by human mammary epithelial cells as revealed by RT/PCR analysis of total RNA from mammary gland and immunohistochemical staining of breast tissue, cultured mammary epithelial cells, and secretory epithelial cells present in human milk. We also verify that immunoreactive leptin is present in whole milk at 30- to 150-fold higher concentrations than skim milk. We propose that leptin is secreted by mammary epithelial cells in milk fat globules, which partition into the lipid portion of breast milk.

Complex mammary tumours in the female dog: a review

2005 ◽  
Vol 72 (S1) ◽  
pp. 90-97 ◽  
Author(s):  
Eva Hellmén
Keyword(s):  
Mammary Gland ◽  
Growth Factors ◽  
Epithelial Cells ◽  
Sex Hormones ◽  
Myoepithelial Cells ◽  
The Body ◽  
Pronounced Effect ◽  
Mammary Tumours ◽  
Luminal Epithelial Cells

Spontaneous mammary tumours are most frequently seen (apart from rodents) in women, female dogs and cats. The mammary gland is the most commonly affected organ for tumours in women and in female dogs. The mammary gland has a similar histology in the different species whereas the number of glands differs as well as the number of interlobular ducts that reach the nipple/teat. The parenchymatous tissue is composed of alveoli that turn into interlobular ducts. The whole ductal tree is outlined by a two-layered epithelium with the luminal epithelial cells adjacent to the lumen and the more sparse myoepithelial cells peripherally located to these. Different proteins such as growth factors regulate the mammary gland, as they do for all tissues in the body. In addition, sex hormones regulate the biology of the mammary gland. Oestrogen has the most pronounced effect on duct growth whereas progesterone promotes growth of the alveoli.

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