scholarly journals ULTRASTRUCTURAL LOCALIZATION OF ACID MUCOSUBSTANCES IN THE MOUSE COLON WITH IRON-CONTAINING STAINS

1966 ◽  
Vol 30 (2) ◽  
pp. 299-315 ◽  
Author(s):  
Mary G. Wetzel ◽  
Bruce K. Wetzel ◽  
Samuel S. Spicer
Keyword(s):  
Ferric Chloride ◽  
Ultrastructural Localization ◽  
Cell Types ◽  
Capillary Endothelium ◽  
Colloidal Iron ◽  
Specific Staining ◽  
Mouse Colon ◽  
Specific Localization ◽  
Cryostat Sections ◽  
Acid Mucosubstances

Selective ultrastructural staining of acid mucosubstances in sites containing histochemically identifiable sulfo- and sialomucins has been obtained in fixed cryostat sections with both ferric chloride and colloidal iron solutions. The rectosigmoid region of mouse colon was fixed in glutaraldehyde, formalin, or phosphate-buffered osmium tetroxide, and 40 µ cryostat sections of this material were treated with 0.1 to 0.4% ferric chloride or with a solution of dialyzed ferric chloride, ammonia, and glycerin. Specific staining depended upon the pH of the iron-containing solutions, and the optimal value was found to be approximately 2.0. Specific localization of acid mucosubstances has been noted in intracellular sites, including globules within colonic goblet cells and "deep crypt" mucous cells, small vesicles of the superficial nongoblet epithelial cells, and Golgi lamellae within each of these cell types. Extracellular material, presumed to be acid mucosubstance, was found on the surface of the epithelial microvilli and on the lumenal surface of capillary endothelium. Similar material formed a reticular network surrounding stromal cells, collagen bundles, and various colonic connective tissue elements.

scholarly journals Single cell transcriptomic analysis of murine lung development on hyperoxia-induced damage

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Maria Hurskainen ◽  
Ivana Mižíková ◽  
David P. Cook ◽  
Noora Andersson ◽  
Chanèle Cyr-Depauw ◽  
...  
Keyword(s):  
Single Cell ◽  
Lung Development ◽  
Alveolar Epithelium ◽  
Cell Types ◽  
Capillary Endothelium ◽  
Stromal Fibroblasts ◽  
Main Driver ◽  
Macrophage Populations ◽  
Induced Changes ◽  
Cellular Compartments

AbstractDuring late lung development, alveolar and microvascular development is finalized to enable sufficient gas exchange. Impaired late lung development manifests as bronchopulmonary dysplasia (BPD) in preterm infants. Single-cell RNA sequencing (scRNA-seq) allows for assessment of complex cellular dynamics during biological processes, such as development. Here, we use MULTI-seq to generate scRNA-seq profiles of over 66,000 cells from 36 mice during normal or impaired lung development secondary to hyperoxia with validation of some of the findings in lungs from BPD patients. We observe dynamic populations of cells, including several rare cell types and putative progenitors. Hyperoxia exposure, which mimics the BPD phenotype, alters the composition of all cellular compartments, particularly alveolar epithelium, stromal fibroblasts, capillary endothelium and macrophage populations. Pathway analysis and predicted dynamic cellular crosstalk suggest inflammatory signaling as the main driver of hyperoxia-induced changes. Our data provides a single-cell view of cellular changes associated with late lung development in health and disease.

scholarly journals CYTOCHEMICAL LOCALIZATION OF ACID MUCOSUBSTANCES WITHIN RABBIT, CHICKEN, FROG AND FISH SKELETAL MUSCLE FIBERS

1969 ◽  
Vol 17 (1) ◽  
pp. 36-46 ◽  
Author(s):  
WILLIAM J. DOUGHERTY
Keyword(s):  
Alcian Blue ◽  
Skeletal Muscles ◽  
Phase Contrast ◽  
Blue Staining ◽  
Polarization Microscopy ◽  
Rabbit Muscle ◽  
Colloidal Iron ◽  
Cytochemical Localization ◽  
High Iron ◽  
Acid Mucosubstances

Skeletal muscles of rabbits, chickens, frogs and fishes were studied by bright field, phase contrast and polarization microscopy after treatment with: (1) Alcian Blue, pH 2.5, (2) Alcian Blue, pH 1.0, (3) low iron diamine, (4) high iron diamine, (5) dialyzed iron, pH 2.0, and (6) colloidal iron, pH 1.5. A prominent aspect of each of the muscles studied was transverse muscle bands which stained with Alcian Blue at pH 2.5, low iron diamine and dialyzed and colloidal iron reagents. Staining of transverse bands was not very intense with these reagents, suggesting that the materials demonstrated occurred in relatively low concentrations. Staining of transverse bands in chicken, frog and fish muscles was observed also after treatment with Alcian Blue at pH 1.0 or with high iron diamine. Staining of rabbit muscle with Alcian Blue at pH 1.0 or with high iron diamine was not detectable even after the most effective fixatives employed in this study, viz., Carony's fixative or buffered HgCl2. Phase contrast and polarization microscopy indicated that the transverse bands stained by the reagents employed corresponded to the I bands in agreement with previous electron cytochemical studies of skeletal muscles treated with dialyzed iron at controlled pH. Methylation for 4 hr at 60°C prevented I band staining with Alcian Blue and dialyzed iron in each of the muscles studied. Treatment of muscle sections with Vibrio cholerae neuraminidase, testicular hyaluronidase or ribonuclease under optimal conditions did not prevent Alcian Blue staining of I bands. Trypsin treatment of muscle sections did not prevent I band staining except when digestion was prolonged to the point that muscle striations were no longer recognizable. Saponification prior to hyaluronidase treatment reduced I band staining in each of the muscles studied. Saponification alone and saponification prior to neuraminidase digestion left I band alcianophilia intact. It was concluded that myofibrillar I bands of rabbit, chicken, frog and fish skeletal muscles contain acid mucosubstances. It was suggested that in each of these organisms an esterified, hyaluronic acid-like molecule is present within the I bands. Sulfated mucosaccharides occur as additional components in the I bands of fish, chicken and frog skeletal muscles; sulfated mucosaccharides apparently are not present in rabbit muscle I bands. Available evidence indicates that sialomucins are not present in the myofibrils of the organisms studied. The possible functional significance of I band acid mucosubstances in vertebrate skeletal muscles was discussed.

Nuclear Morphology and the Ultra-Structural Localization of Deoxyribonucleic Acid Synthesis during Interphase

1969 ◽  
Vol 4 (3) ◽  
pp. 569-582
Author(s):  
GILLIAN R. MILNER
Keyword(s):  
Epithelial Cells ◽  
Dna Synthesis ◽  
Deoxyribonucleic Acid ◽  
Ultrastructural Localization ◽  
Cell Types ◽  
Acid Synthesis ◽  
Lung Fibroblasts ◽  
Nuclear Morphology ◽  
Structural Localization ◽  
Electron Microscope Autoradiography

The ultrastructural localization of deoxyribonucleic acid (DNA) synthesis was studied by electron-microscope autoradiography in human transforming lymphocytes, embryonic lung fibroblasts, epithelial cells and normoblasts. Euchromatin was found to be active in DNA synthesis in all cell types studied, whereas heterochromatin was inactive. However, DNA synthesis was also prominent in the regions where heterochromatin was thought to be decondensing to form euchromatin. Analysis of sequential changes in nuclear morphology of the transforming lymphocyte suggested that there is decondensation of heterochromatin during the S-phase until none is left. In nuclei with no heterochromatin a prominent localization of DNA synthesis was at the nuclear membrane. This sequence of complete decondensation of heterochromatin also seemed likely for fibroblasts and epithelial cells. Normoblasts however showed no stage where the nucleus was wholly euchromatic and it is suggested that in this cell decondensation of heterochromatin for replication is localized and transient.

Abstract 1433: The Cardioregulatory Peptide Apelin is Preferentially Expressed by Endothelial Cells and Upregulated in Myocardial Disease States

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Ramendra K Kundu ◽  
Ahmad Y Sheikh ◽  
Michael Y Ho ◽  
Hyung J Chun ◽  
Diem T Huynh ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Reporter Gene ◽  
Fold Increase ◽  
Cell Types ◽  
Heart Tissue ◽  
Capillary Endothelium ◽  
Lacz Gene ◽  
Lacz Reporter ◽  
Rt Pcr ◽  
Disease States

Introduction: The APJ receptor and its ligand, apelin, comprise a homeostatic, cardio-regulatory pathway. Although cardiac apelin expression levels are altered in humans with cardiac failure, the cell type responsible for apelin expression and modulation in disease states remains unknown. Hypothesis: Apelin production is restricted to the endothelial compartment and is upregulated in states of cardiovascular stress. Methods: Transgenic apelin-LacZ reporter mice (SVJ background) were created by insertion of the nuclear localizing bacterial LacZ gene immediately downstream of the apelin promoter. Mice (n=12) were randomized to left anterior descending coronary artery (LAD) ligation, thoracic aortic constriction (TAC) or sham groups. Hearts were harvested 3 and 8 weeks post-TAC or LAD ligation, respectively. Localization of apelin expression was determined by Xgal staining. Endothelial phenotype of lacZ positive cells was confirmed by CD31 co-staining. Apelin expressing cells were quantified by histology. Apelin reporter results were confirmed by quantitating apelin expression in WT animals following LAD ligation (n=11) or sham (n=11) procedure by RT-PCR. Results: Extensive immunohistochemistry studies of heart tissue revealed lacZ reporter gene expression to be restricted to the coronary venous and capillary endothelium, with no expression by cardiomyocytes. Following both LAD ligation and TAC, the number of LacZ-apelin (+) endothelial cells significantly increased (p<0.002) in all chambers of the heart (Table ), with no evidence of apelin expression by other cell types. Evaluation of WT hearts by RT-PCR for the apelin gene confirmed the reporter gene findings with 1.3±0.3 fold increase (p<0.05) of apelin expression induced by LAD ligation compared to sham. Conclusions: Apelin is primary expressed by endothelial cells within the heart and is upregulated in response to myocardial stress. Apelin-LacZ Expressing Endothelial Cells are Increased Following Myocardial Injury

Lung tissue volume changes induced by hypertonic NaCl: morphometric evaluation

1981 ◽  
Vol 51 (6) ◽  
pp. 1443-1450 ◽  
Author(s):  
D. Wangensteen ◽  
H. Bachofen ◽  
E. R. Weibel
Keyword(s):  
Lung Tissue ◽  
Alveolar Epithelium ◽  
Cell Types ◽  
Ringer Solution ◽  
Capillary Endothelium ◽  
Vascular Perfusion ◽  
Tissue Samples ◽  
Membrane Area ◽  
Capillary Lumen ◽  
Osmotic Effect

Lung tissue was examined to determine how the volumes of alveolar septum components change when NaCl is added to the vascular perfusate, increasing the osmolarity by 70 mosM. Isolated rabbit lungs were perfused with Ringer solution containing dextran, either with or without added NaCl, and fixed by vascular perfusion. Tissue samples from both “control” and “hypertonic” lungs, prepared for electron microscopy, were examined using established morphometric procedures. Volumes of septal cells, interstitial space, capillary lumen, surface-lining layer, and endothelial and epithelial areas were measured, all normalized against the endothelium basement-membrane area. Results showed that hypertonic NaCl caused a reduction in total cell and surface-lining layer volumes but no change in interstitial or capillary lumen volumes. This supports the hypothesis that small molecules have no osmotic effect across the pulmonary capillary endothelium but do cause a fluid flux from cells and across the alveolar epithelium. Areas and volume measurements for different septal cell types suggest a heterogeneous response: epithelial cells showed significant decreases and endothelial cells changed little, if at all.

scholarly journals Ultrastructural localization of lactoferrin and myeloperoxidase in human neutrophils by immunogold

Blood ◽  
1985 ◽  
Vol 65 (2) ◽  
pp. 423-432 ◽  
Author(s):  
E Cramer ◽  
KB Pryzwansky ◽  
JL Villeval ◽  
U Testa ◽  
J Breton-Gorius
Keyword(s):  
Human Peripheral Blood ◽  
Ultrastructural Localization ◽  
Human Neutrophils ◽  
Cell Fractionation ◽  
Thin Sections ◽  
The Matrix ◽  
Blood Neutrophils ◽  
Microscopic Studies ◽  
Specific Localization ◽  
Monospecific Antibodies

Abstract Colloidal gold was used as a marker for immunoelectron microscopy to localize lactoferrin (LF) and myeloperoxidase (MPO) in human peripheral blood neutrophils. Cells were reacted with monospecific antibodies against LF or MPO and then with gold-labeled antiglobulin. MPO cytochemistry was also associated with immunologic detection of LF. Immunologic labeling of thin sections after embedding in glycol methacrylate gave good ultrastructural morphology and specific localization of both proteins. MPO was detected in the large azurophil granules, whereas LF was consistently localized in the matrix of another population of morphologically distinct granules, smaller and more numerous than azurophil granules. When cytochemical detection of MPO was coupled with immunologic detection of LF, LF was observed in the population of MPO-negative granules, which were identified as specific. This was confirmed on cells that were permeabilized with saponin and stained for LF and MPO before embedding. No other neutrophil organelles displayed labeling for LF; other blood cells also were unreactive for LF. In the bone marrow, myeloblast and promyelocyte granulations were not stained and LF-containing granules appeared at the myelocyte stage. In conclusion, we confirm previous biochemical and light microscopic studies by ultrastructural demonstration of LF and MPO in two categories of granules, the specific and azurophil granules, respectively. The method described in this article avoids disruption caused by cell fractionation procedures. In the future, other intragranular proteins can be localized by a similar approach.

scholarly journals Methods to Enhance Signal Using Isotopic In Situ Hybridization

2002 ◽  
Vol 50 (8) ◽  
pp. 1031-1037 ◽  
Author(s):  
Betty Ky ◽  
Paul J. Shughrue
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Cell Types ◽  
Oxytocin Receptor ◽  
Specific Cell ◽  
Rat Hypothalamus ◽  
Low Levels ◽  
Tuberoinfundibular Peptide ◽  
Cryostat Sections

Isotopic in situ hybridization (ISH) has been established as a uniquely powerful tool for the study of gene expression in specific cell types. This technique allows the visualization and quantification of gene expression and gene expression changes in cells. In our study of biological and molecular phenomena, we have increasingly encountered the need to detect small changes in gene expression as well as genes of low abundance, such as the oxytocin receptor (OTR) and the tuberoinfundibular peptide of 39 residues (Tip39). To increase the sensitivity of isotopic ISH for detection of rare mRNAs, we performed ISH on cryostat sections of rat hypothalamus and thalamus with 35S-labeled riboprobes and amplified the signal by hybridizing over 2 nights as well as labeling the probe with both [35S]-UTP and [35S]-ATP. These two methods of enhancement independently and in combination demonstrated a dramatic increase in signal, allowing the visualization of low levels of gene expression previously undetectable by conventional methods.

scholarly journals Reciprocal expression of 17α-hydroxylase-C17,20-lyase and aromatase cytochrome P450 during bovine trophoblast differentiation: a two-cell system drives placental oestrogen synthesis

Reproduction ◽  
2006 ◽  
Vol 131 (4) ◽  
pp. 669-679 ◽  
Author(s):  
G Schuler ◽  
G R Özalp ◽  
B Hoffmann ◽  
N Harada ◽  
P Browne ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Cell Types ◽  
Staining Intensity ◽  
Giant Cells ◽  
Cell System ◽  
Cellular Level ◽  
Specific Staining ◽  
Cytoplasmic Staining ◽  
Distinct Cell ◽  
Trophoblast Giant Cells

No definitive information is yet available on the steroidogenic capacity of the two morphologically distinct cell types forming the bovine trophoblast, the uninucleated trophoblast cells (UTCs) and the trophoblast giant cells (TGCs). Hence, in order to localise 17α-hydroxylase-C17,20-lyase (P450c17) on a cellular level and to monitor its expression as a function of gestational age, placentomes from pregnant (days 80–284; n = 19), prepartal (days 273–282; 24–36 h prior to the onset of labour; n = 3) and parturient cows (n = 5) were immunostained for P450c17 using an antiserum against the recombinant bovine enzyme. At all stages investigated, P450c17 was exclusively found in the UTCs of chorionic villi (CV), where staining was ubiquitous between days 80 and 160, but was largely restricted to primary CV and the branching sites of secondary CV between days 160 and 240. Thereafter, a distinct ubiquitous staining reoccurred in the UTCs of all CV in late pregnant, prepartal and parturient animals. Using an antiserum against human aromatase cytochrome P450 (P450arom), specific cytoplasmic staining was observed in TGCs. In placentomes from pregnant cows, staining intensity was higher in mature compared with immature TGCs and was more pronounced in the trophoblast covering big stem villi compared with the trophoblast at other sites of the villous tree. In placentomes of a parturient cow, specific staining was only found in mature TGCs that survived the normal, but substantial, prepartal decline in TGC numbers. These results clearly showed that bovine UTCs and TGCs exhibit different steroidogenic capacities, constituting a ‘two-cell’ organisation for oestrogen synthesis. P450c17 expression appears to be quickly down-regulated and P450arom is up-regulated when UTCs enter the TGC differentiation pathway.

Immunocytochemical localization of the nuclear 3,5,3'-triiodothyronine receptor in the adult rat: liver, kidney, heart, lung and spleen

1989 ◽  
Vol 120 (4) ◽  
pp. 451-458 ◽  
Author(s):  
M. Luo ◽  
R. Faure ◽  
Y. A. Tong ◽  
J. H. Dussault
Keyword(s):  
Ascitic Fluid ◽  
Cell Types ◽  
Type Ii ◽  
Myocardial Cells ◽  
Cell Nuclei ◽  
Macrophage Cell ◽  
Specific Staining ◽  
Adult Rat ◽  
Type Ii Pneumocytes ◽  
Mature Lymphocyte

Abstract. A monoclonal antibody was used for the localization of the nuclear T3 receptor in different tissues of the adult rat: the liver, kidney, heart, lung, spleen, testis, and pituitary. In the liver, the immunoreactivity was found uniformly distributed in the nuclei of hepatocytes. Sections incubated with a control ascitic fluid or with the same ascitic fluid pre-adsorbed with purified receptor showed no specific staining. In the kidney, the immunoreactivity was higher in the epithelial cell of the proximal convoluted tubes and juxtaglomerular cells. In the heart, only the myocardial cells were stained. In the lung, the immunoreactivity was confined to type II pneumocytes and alveolar macrophages. In the spleen, only a few mature lymphocyte and macrophage cell nuclei were stained. These results show that: 1) the abundance of the nuclear T3 correlates with previous studies using hormone binding techniques; 2) the nuclear T3 receptor is selectively located in certain cell types, which possess a precise local function.

scholarly journals Electron histochemistry and ultrastructural localization of carbohydrate-containing substances in the sheath of Volvox.

1976 ◽  
Vol 24 (5) ◽  
pp. 668-673 ◽  
Author(s):  
M D McCracken ◽  
W J Barcellona
Keyword(s):  
Periodic Acid ◽  
Ultrastructural Localization ◽  
Ruthenium Red ◽  
Cationic Dyes ◽  
External Layer ◽  
Colloidal Iron ◽  
Electron Histochemistry ◽  
Reactive Groups ◽  
Sheath Material

The location and characteristics of carbohydrate-containing structures within the intact sheath of Volvox were studied by 3,3'-diaminobenzidine tetrahydrochloride-osmium, colloidal iron, colloidal thorium, ruthenium red and periodic acid-silver methenamine staining. The sheath consists of external and internal fibrillar layers separated by a tripartite structure. The external layer reacts positively with 3,3'-diaminobenzidine tetrahydrochloride, colloidal iron, colloidal thorium and ruthenium red, indicating that it contains acid mucosaccharides. Staining in the external layer is abolished by Ba(OH)2 treatment. The tripartite structure and internal fibrillar layer contain periodic acid reactive groups which do not occur in the external layer. Under certain conditions, reactions between the cationic dyes and the internal material were also observed. It is postulated that the internal matrix of the sheath contains glycoproteins or a mixture of acid mucosaccharides and glycoproteins. Possible functions of the sheath material are discussed.

Sign in / Sign up

Export Citation Format

Share Document