scholarly journals Immunoelectron microscopic localization of 3beta-hydroxysteroid dehydrogenase and type 5 17beta-hydroxysteroid dehydrogenase in the human prostate and mammary gland

2001 ◽  
Vol 26 (1) ◽  
pp. 11-19 ◽  
Author(s):  
G Pelletier ◽  
V Luu-The ◽  
M El-Alfy ◽  
S Li ◽  
F Labrie
Keyword(s):  
Endothelial Cells ◽  
Mammary Gland ◽  
Epithelial Cells ◽  
Blood Vessels ◽  
Sex Steroids ◽  
Cell Types ◽  
Hydroxysteroid Dehydrogenase ◽  
Human Prostate ◽  
Peripheral Tissues ◽  
Gold Particles

The subcellular distribution of steroidogenic enzymes has so far been studied mostly in classical endocrine glands and in the placenta. In the peripheral intracrine organs which synthesize sex steroids there is no indication about the organelles which contain the enzymes involved in steroid biosynthesis. We have thus investigated the subcellular localization of two enzymes involved in the production of sex steroids, namely 3beta-hydroxysteroid dehydrogenase (3beta-HSD) and type 5 17beta-hydroxysteroid dehydrogenase (17beta-HSD). Using specific antibodies to these enzymes, we conducted immunoelectron microscopic studies in two peripheral tissues, namely the human prostate and mammary gland. In the prostate, immunolabelling for both 3beta-HSD and type 5 17beta-HSD was detected in the basal cells of the tube-alveoli as well as in fibroblasts and endothelial cells lining the blood vessels. In all the labelled cell types, the gold particles were distributed throughout the cytoplasm. No obvious association with any specific organelle could be observed, although some concentration of gold particles was occasionally found over bundles of microfilaments. In mammary gland sections immunolabelled for 3beta-HSD or type 5 17beta-HSD localization, labelling was observed in the cytoplasm of the secretory epithelial cells in both the acini and terminal ducts. Immunolabelling was also found in the endothelial cells as well as in fibroblasts in stroma and blood vessels. The gold particles were not detected over any organelles, except with the occasional accumulation of gold particles over microfilaments. The present data on the localization of two steroidogenic enzymes leading to the synthesis of testosterone indicate that these enzymes are located not only in epithelial cells but also in stromal and endothelial cells in both tissues studied. The absence of any association of the enzymes with membrane-bound organelles appears as a common finding in the reactive cell types of two peripheral tissues.

scholarly journals BIM deficiency differentially impacts the function of kidney endothelial and epithelial cells through modulation of their local microenvironment

2012 ◽  
Vol 302 (7) ◽  
pp. F809-F819 ◽  
Author(s):  
Nader Sheibani ◽  
Margaret E. Morrison ◽  
Zafer Gurel ◽  
SunYoung Park ◽  
Christine M. Sorenson
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Epithelial Cells ◽  
Cell Types ◽  
Proper Function ◽  
Causative Role ◽  
Structural Support ◽  
Cell Adhesion And Migration ◽  
And Migration ◽  
Endothelial Nitric Oxide

The extracellular matrix (ECM) acts as a scaffold for kidney cellular organization. Local secretion of the ECM allows kidney cells to readily adapt to changes occurring within the kidney. In addition to providing structural support for cells, the ECM also modulates cell survival, migration, proliferation, and differentiation. Although aberrant regulation of ECM proteins can play a causative role in many diseases, it is not known whether ECM production, cell adhesion, and migration are regulated in a similar manner in kidney epithelial and endothelial cells. Here, we demonstrate that lack of BIM expression differentially impacts kidney endothelial and epithelial cell ECM production, migration, and adhesion, further emphasizing the specialized role of these cell types in kidney function . Bim −/− kidney epithelial cells demonstrated decreased migration, increased adhesion, and sustained expression of osteopontin and thrombospondin-1 (TSP1). In contrast, bim −/− kidney endothelial cells demonstrated increased cell migration, and decreased expression of osteopontin and TSP1. We also observed a fivefold increase in VEGF expression in bim −/− kidney endothelial cells consistent with their increased migration and capillary morphogenesis. These cells also had decreased endothelial nitric oxide synthase activity and nitric oxide bioavailability. Thus kidney endothelial and epithelial cells make unique contributions to the regulation of their ECM composition, with specific impact on adhesive and migratory properties that are essential for their proper function.

scholarly journals The absence of H-2 antigens from mouse pancreatic beta-cells demonstrated by immunoferritin labeling.

1979 ◽  
Vol 150 (1) ◽  
pp. 1-9 ◽  
Author(s):  
E L Parr
Keyword(s):  
Endothelial Cells ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Beta Cell ◽  
Pancreatic Duct ◽  
Beta Cells ◽  
Pancreatic Beta Cells ◽  
Acinar Cells ◽  
Cell Types ◽  
Capillary Endothelial Cells

Islets of Langerhans were isolated from mouse pancreases and fixed in periodatelysine-paraformaldehyde. The fixed islets were then dissociated with trypsin and EDTA to yield cell suspensions that contained mainly four cell types; beta-cells, capillary endothelial cells, acinar cells, and pancreatic duct epithelial cells. The nonislet cells were probably associated wtih the surface of the isolated islets. The H-2 antigens of the dissociated pancreatic cells were labeled with an immunoferritin technique. Pancreatic duct epithelial cells showed specific ferritin labeling on their lateral cell membranes but not on apical microvillus membranes. Acinar cells were also labeled on lateral membranes, and the capillary endothelial cells were labeled on both the luminal and albuminal aspects of their surface membranes. In contrast, pancreatic beta-cells were unlabeled. The number of ferritin molecules per unit length of beta-cell membrane was essentially the same on cells from the antigenic strain and the congeneic control strain, and was about 200-fold less than on the labeled pancreatic duct epithelial cell lateral membranes. Pancreatic beta-cells are therefore one of six known epithelial cell types on which H-2 antigens can not be detected by immunoferritin labeling. The apparent absence of H-2 antigens from these cells suggests a study of the viability of beta-cells in allografts of dissociated islet cells, in which the beta-cell would not be in contact with antigenic cells. Such studies might lead to a new approach to the control of diabetes mellitus by transplantation.

scholarly journals Endothelial Claudin

1999 ◽  
Vol 147 (1) ◽  
pp. 185-194 ◽  
Author(s):  
Kazumasa Morita ◽  
Hiroyuki Sasaki ◽  
Mikio Furuse ◽  
Shoichiro Tsukita
Keyword(s):  
Endothelial Cells ◽  
Epithelial Cells ◽  
Blood Vessels ◽  
Polyclonal Antibody ◽  
Polyclonal Antibodies ◽  
Transmembrane Protein ◽  
Velo Cardio Facial Syndrome ◽  
The Brain ◽  
Specific Polyclonal Antibody

Tight junctions (TJs) in endothelial cells are thought to determine vascular permeability. Recently, claudin-1 to -15 were identified as major components of TJ strands. Among these, claudin-5 (also called transmembrane protein deleted in velo-cardio-facial syndrome [TMVCF]) was expressed ubiquitously, even in organs lacking epithelial tissues, suggesting the possible involvement of this claudin species in endothelial TJs. We then obtained a claudin-6–specific polyclonal antibody and a polyclonal antibody that recognized both claudin-5/TMVCF and claudin-6. In the brain and lung, immunofluorescence microscopy with these polyclonal antibodies showed that claudin-5/TMVCF was exclusively concentrated at cell–cell borders of endothelial cells of all segments of blood vessels, but not at those of epithelial cells. Immunoreplica electron microscopy revealed that claudin-5/TMVCF was a component of TJ strands. In contrast, in the kidney, the claudin-5/TMVCF signal was restricted to endothelial cells of arteries, but was undetectable in those of veins and capillaries. In addition, in all other tissues we examined, claudin-5/TMVCF was specifically detected in endothelial cells of some segments of blood vessels, but not in epithelial cells. Furthermore, when claudin-5/TMVCF cDNA was introduced into mouse L fibroblasts, TJ strands were reconstituted that resembled those in endothelial cells in vivo, i.e., the extracellular face–associated TJs. These findings indicated that claudin-5/TMVCF is an endothelial cell–specific component of TJ strands.

scholarly journals Development of mouse mammary gland: identification of stages in differentiation of luminal and myoepithelial cells using monoclonal antibodies and polyvalent antiserum against keratin.

1986 ◽  
Vol 34 (8) ◽  
pp. 1037-1046 ◽  
Author(s):  
A Sonnenberg ◽  
H Daams ◽  
M A Van der Valk ◽  
J Hilkens ◽  
J Hilgers
Keyword(s):  
Monoclonal Antibodies ◽  
Mammary Gland ◽  
Epithelial Cells ◽  
Basement Membrane ◽  
Cell Types ◽  
Mouse Mammary Gland ◽  
Type I ◽  
Basal Cells ◽  
Alveolar Cells ◽  
Luminal Type

The development of the mouse mammary gland was studied immunohistochemically using monoclonal antibodies against cell surface and basement membrane proteins and a polyclonal antibody against keratin. We have identified three basic cell types: basal, myoepithelial, and epithelial cells. The epithelial cells can be subdivided into three immunologically related cell types: luminal type I, luminal type II, and alveolar cells. These five cell types appear at different stages of mammary gland development and have either acquired or lost one of the antibody-defined antigens. The cytoplasmic distribution of several of these antigens varied according to the location of the cells within the mammary gland. Epithelial cells which did not line the lumen expressed antigens throughout the cytoplasm. These antigens were demonstrated on the apical site in situations where the cells lined the lumen. One antigen became increasingly basolateral as the cells became attached to the basement membrane. The basal cells synthesize laminin and deposit it at the cell base. They are present in endbuds and ducts and are probably the stem cells of the mammary gland. Transitional forms have been demonstrated which developmentally link these cells with both myoepithelial and (luminal) epithelial cells.

scholarly journals Single-cell RNA-Aequencing Reveals Novel Myofibroblasts with Epithelial Cell-Like Features in the Mammary Gland of Dairy Cattle

2020 ◽  
Author(s):  
Fengfei Gu ◽  
Jiajin Wu ◽  
Senlin Zhu ◽  
Teresa G. Valencak ◽  
Jian-Xin Liu ◽  
...  
Keyword(s):  
Mammary Gland ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Dairy Cattle ◽  
Single Cell ◽  
Dairy Product ◽  
Cell Types ◽  
Marker Genes ◽  
New Findings ◽  
Luminal Cells

Abstract Background: Cow’s milk is a highly-nutritious dairy product that is widely consumed worldwide. It is secreted by the developed mammary gland (MG) of dairy cattle. However, a comprehensive understanding of cell-type diversity and cell function within bovine MG is lacking. In the current study, we used single-cell RNA sequencing to investigate the transcriptome of 24,472 high-quality MG cells isolated from newborn and adult cows. Results: Unbiased clustering analysis revealed the existence of 24 cell types, which could be divided into four categories: 9 immune, 3 epithelial, 9 fibroblast, and 3 endothelial cell types. Other cell subtypes were further identified based on re-clustering and pseudotemporal reconstruction of epithelial cells that included 3 mature luminal epithelial, 1 intermediate, and 2 progenitor cell subtypes. The individual top marker genes of these 3 mature luminal epithelial cell subtypes (L0, L1, and L5) were APOA1, STC2, and PTX3, which were further validated using immunofluorescence. Based on functional analysis, the L0, L1, and L5 cell subtypes were all involved in the upregulation of lipid metabolism, protein and hormone metabolism, and the immune response, respectively. Furthermore, we discovered a novel myofibroblast that expresses COL1A1 and CSN3, has visible epithelial-like characteristics, and shows the potential to differentiate into luminal epithelial cells, especially immune-sensing luminal cells (L5). Conclusions: We constructed the first single-cell atlas of the dairy cow MG, and our new findings of epithelial-like myofibroblast cells and their differentiation trajectories into luminal cells may provide novel insights into the development and lactogenesis in dairy cattle MGs.

Ezrin is concentrated in the apical microvilli of a wide variety of epithelial cells whereas moesin is found primarily in endothelial cells

1993 ◽  
Vol 105 (4) ◽  
pp. 1025-1043 ◽  
Author(s):  
M. Berryman ◽  
Z. Franck ◽  
A. Bretscher
Keyword(s):  
Endothelial Cells ◽  
Epithelial Cells ◽  
Cultured Cells ◽  
Cell Types ◽  
Cytoskeletal Proteins ◽  
Specific Cell ◽  
Apical Surface ◽  
Tissue Sections ◽  
Differentiated Cells

Ezrin and moesin are two cytoskeletal proteins originally purified from human placenta that are 74% identical in overall protein sequence. They are believed to be membrane-cytoskeletal linking proteins because they share sequence homology with erythrocyte band 4.1 and colocalize with actin specifically in microvilli and membrane ruffles in cultured cells. To determine if ezrin and moesin share similar distributions in vivo, we studied their localizations with respect to F-actin in tissue sections. Surprisingly, ezrin and moesin exhibited very different cellular distributions. Ezrin was highly concentrated and colocalized with actin on the apical surface of many epithelial cell types. During enterocyte differentiation, the pattern of expression and redistribution of ezrin was consistent with it performing a role in microvillus assembly. Immunoelectron microscopy in differentiated cells revealed that ezrin was restricted mainly to the plasma membrane of microvilli and other actin-rich surface projections. Moesin was found in endothelial cells and was also enriched in the apical microvilli of a restricted set of epithelial cells. All polarized cell types with abundant microvilli contained one or both proteins, suggesting that ezrin and moesin perform related functions. However, the differential expression of ezrin and moesin indicates that they have distinct properties, which are uniquely adapted to specific cell types.

scholarly journals Antibodies to mouse lung capillary endothelium.

1988 ◽  
Vol 36 (7) ◽  
pp. 741-749 ◽  
Author(s):  
M C Rorvik ◽  
D P Allison ◽  
J A Hotchkiss ◽  
H P Witschi ◽  
S J Kennel
Keyword(s):  
Endothelial Cells ◽  
Cell Types ◽  
Interstitial Cells ◽  
Mouse Lung ◽  
Capillary Endothelium ◽  
Specific Cell ◽  
Alveolar Wall ◽  
Gold Particles ◽  
Capillary Endothelial Cells ◽  
Quantitative Analyses

We are interested in developing monoclonal antibodies (MoAbs) that recognize specific cell types in the lung of BALB/c mice. Normal mouse lung homogenate was used to immunize F344 rats and hybridomas were produced by fusion of rat spleen cells with mouse myeloma SP 2/0. Two hybridomas were selected which produced MoAbs active in immunohistochemistry of lung cells. MoAb 273-34A and 411-201B both show extensive peroxidase staining of capillary endothelial cells within alveolar walls of lungs at the light microscopic level. To demonstrate cell specificity, immunoelectron microscopy with gold-labeled antibody was performed. Lightly fixed lungs were frozen and thin-sectioned before staining with MoAb and 5-nm gold particles coupled to secondary antibody. Quantitative analyses of these cryosections show that both antibodies, used at optimal concentrations, are specific for binding to capillary endothelial cells. More than 95% of the gold particles are associated with capillary endothelial cells on the thin side of the alveolar wall. When capillaries adjoined thick septa containing interstitial cells, about two thirds of the gold particles were associated with endothelial cells and about one quarter with interstitial cells. These MoAbs should be useful in studying the role of endothelial cells in toxic lung injury.

scholarly journals Transcriptome Analysis of Neisseria meningitidis during Infection

2003 ◽  
Vol 185 (1) ◽  
pp. 155-164 ◽  
Author(s):  
Guido Dietrich ◽  
Sebastian Kurz ◽  
Claudia Hübner ◽  
Christian Aepinus ◽  
Stephanie Theiss ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Epithelial Cells ◽  
Neisseria Meningitidis ◽  
Dna Microarrays ◽  
Environmental Changes ◽  
Cell Types ◽  
Open Reading Frames ◽  
Meningococcal Infection ◽  
Considerable Proportion ◽  
Transcriptional Level

ABSTRACT Neisseria meningitidis is the cause of septicemia and meningococcal meningitis. During the course of infection, N. meningitidis encounters multiple environments within its host, which makes rapid adaptation to environmental changes a crucial factor for neisserial pathogenicity. Employing oligonucleotide-based DNA microarrays, we analyzed the transcriptome of N. meningitidis during two key steps of meningococcal infection, i.e., the interaction with epithelial cells (HeLa cells) and endothelial cells (human brain microvascular endothelial cells). Seventy-two genes were differentially regulated after contact with epithelial cells, and 48 genes were differentially regulated after contact with endothelial cells, including a considerable proportion of well-known virulence genes. While a considerable number of genes were in concordance between bacteria adherent to both cell types, we identified several open reading frames that were differentially regulated in only one system. The data obtained with this novel approach may provide insight into the pathogenicity mechanisms of N. meningitidis and could demonstrate the importance of gene regulation on the transcriptional level during different stages of meningococcal infection.

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