A histochemical study of sulphated glycosaminoglycans associated with the somites of the chick embryo

Development ◽  
1973 ◽  
Vol 29 (1) ◽  
pp. 197-208
Author(s):  
M. J. O'Hare
Keyword(s):  
Chick Embryo ◽  
Basement Membrane ◽  
Alcian Blue ◽  
Chondroitin Sulphate ◽  
Matrix Material ◽  
Paraxial Mesoderm ◽  
Membrane Material ◽  
Sulphated Glycosaminoglycans ◽  
Sulphated Glycosaminoglycan ◽  
Chondroitin Sulphate A

A histochemical analysis has been made of sulphated glycosaminoglycans (mucopolysaccharides) associated with chick embryo somites before and after the onset of overt cartilage differentiation. The sulphated glycosaminoglycans were distinguished and resolved into different types by the use of alcian blue at low pH and alcian blue ‘critical electrolyte concentration’ staining combined with hyaluronidase digestion. The newly formed somites are bounded on their dorsal, ventral, and medial surfaces by basement membrane material as they are delimited from the unsegmented paraxial mesoderm Such epithelial basement membrane material, which was first detected in association with the epiblast/mesoderm boundary in the stage-4 embryo, was found to contain a major chondroitin sulphate A/C fraction and a minor chondroitin sulphate B fraction. The notochord sheath contained similar sulphated glycosaminoglycans. Sulphated glycosaminoglycans were first detected between cells of the somite ‘core’ at stage 14 and were subsequently seen to accumulate around the cells of the developing sclerotome and later around cells of the dermatome; the myotome was devoid of such material at these stages (stage 14–20). These pre-cartilaginous sulphated glycosaminoglycans were also of the chondroitin sulphate A/C plus chondroitin sulphate B types. In contrast, the matrix material of newly forming vertebral cartilage, which was first seen in the anterior region of stage 21 embryos, was distinguished by its lack of a hyaluronidase-resistant sulphated glycosaminoglycan component, and therefore presumably contained only chondroitin sulphates A/C. Much later in development (after stage 33) small amounts of sulphated glycosaminoglycan with the staining properties of keratan sulphate were found in the perichordal and subperichondrial regions of the vertebral cartilage.

Chondrogenesis in chick embryo somites grafted with adjacent and heterologous tissues

Development ◽  
1972 ◽  
Vol 27 (1) ◽  
pp. 229-234
Author(s):  
M. J. O'Hare
Keyword(s):  
Chick Embryo ◽  
Basement Membrane ◽  
Membrane Material ◽  
Grafting Technique ◽  
Cartilage Differentiation ◽  
Sulphated Glycosaminoglycans ◽  
Embryonic Ectoderm

A variety of heterologous tissues have been tested for the ability to promote cartilage differentiation in isolated chick-embryo somites, using a modified chorioallantoic grafting technique. Of the 12 tissues tested only 3- and 4-day embryonic ectoderm promoted somite chondrogenesis in somites that fail to chondrify when grafted in isolation. This activity of ectoderm was evident in grafts of somites isolated with adjacent ectoderm, and in grafts of somites recombined with ectoderm derived from several sources. Four-day embryonic limbbud ectoderm, including the apical ridge, was capable of promoting somite chondrogenesis, but to no greater extent than dorsal trunk ectoderm of the same age. It is suggested that the ability of embryonic ectoderm to promote cartilage differentiation in isolated somites is associated with its ability to synthesize basement membrane material (sulphated glycosaminoglycans and collagen), in association with adjacent somite mesoderm.

scholarly journals Neural 16S acetylcholinesterase is solubilized by heparin

1983 ◽  
Vol 215 (1) ◽  
pp. 201-204 ◽  
Author(s):  
J C Torres ◽  
M I Behrens ◽  
N C Inestrosa
Keyword(s):  
Sciatic Nerve ◽  
Esterase Activity ◽  
Ache Activity ◽  
Chondroitin Sulphate ◽  
Sedimentation Analysis ◽  
Rat Sciatic Nerve ◽  
Sulphated Glycosaminoglycans ◽  
Sulphated Glycosaminoglycan ◽  
Solubilizing Effect

The effect of heparin, a sulphated glycosaminoglycan, on the solubilization of rat sciatic-nerve acetylcholinesterase (acetylcholine acetylhydrolase; AChE; EC 3.1.1.7) was studied. It was found that heparin solubilized esterase activity from ligated nerves. Sedimentation analysis revealed this activity to be mainly the 16S form. Chondroitin sulphate did not solubilize AChE activity, and protamine eliminated the solubilizing effect. Our results suggest the involvement of sulphated glycosaminoglycans in the intra-axonal localization and transport of 16S AChE.

scholarly journals Polyacrylamide-gel electrophoresis and Alcian Blue staining of sulphated glycosaminoglycan oligosaccharides

1984 ◽  
Vol 221 (3) ◽  
pp. 707-716 ◽  
Author(s):  
M K Cowman ◽  
M F Slahetka ◽  
D M Hittner ◽  
J Kim ◽  
M Forino ◽  
...  
Keyword(s):  
Alcian Blue ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Unit Structure ◽  
Electrophoretic Patterns ◽  
Sulphated Glycosaminoglycans ◽  
Sulphated Glycosaminoglycan ◽  
Testicular Hyaluronidase

Oligosaccharide fragments of glycosaminoglycans may be separated for rapid analysis by electrophoresis through a 10% polyacrylamide matrix. An extensive ladder-like set of bands is observed for partial testicular hyaluronidase digests of chondroitin 4- or 6-sulphate, and for dermatan sulphate. Co-electrophoresis of purified oligosaccharides has established that the major bands of these patterns represent fragments differing in chain length by one disaccharide unit, with the smallest fragments having the greatest mobility. Additional minor bands, representing heterogeneity in the repeating unit structure, are also observed. There are slight differences in the mobilities of oligosaccharides derived from the three major types of sulphated glycosaminoglycans. Alcian Blue is employed for visualization of the digest fragments. Sample loads of 5-10 micrograms per band appear optimum. The smallest oligosaccharide which may be stained by this method is the hexasaccharide. After consideration of this effect, a good correlation is found to exist between densitometric scans of the gel-electrophoretic patterns and gel-filtration chromatographic profiles based on uronic acid concentration.

vitreal cells and specialized phagocytes in the chick embryo retina: A TEM and SEM study

1990 ◽  
Vol 48 (3) ◽  
pp. 330-331
Author(s):  
M.A. Cuadros ◽  
M.J. Martinez-Guerrero ◽  
A. Rios
Keyword(s):  
Cell Death ◽  
Plasma Membrane ◽  
Acid Phosphatase ◽  
Chick Embryo ◽  
Basement Membrane ◽  
Sem Images ◽  
Intimate Contact ◽  
Cellular Processes ◽  
Sem Study ◽  
Free Cells

In the chick embryo retina (days 3-4 of incubation), coinciding with an increase in cell death, specialized phagocytes characterized by intense acid phosphatase activity have been described. In these preparations, all free cells in the vitreal humor (vitreal cells) were strongly labeled. Conventional TEM and SEM techniques were used to characterize them and attempt to determine their relationship with retinal phagocytes.Two types of vitreal cells were distinguished. The first are located at some distance from the basement membrane of the neuroepithelium, and are rounded, with numerous vacuoles and thin cytoplasmic prolongations. Images of exo- and or endocytosis were frequent; the cells showed a well-developed Golgi apparatus (Fig. 1) In SEM images, the cells was covered with short cellular processes (Fig. 3). Cells lying parallel to or alongside the basement membrane are elongated. The plasma membrane is frequently in intimate contact with the basement membrane. These cells have generally a large cytoplasmic expansion (Fig. 5).

Ultrastructural evidence of transplacental transport of immunoglobulin G in bitches

Reproduction ◽  
2000 ◽  
pp. 315-326 ◽  
Author(s):  
MH Stoffel ◽  
AE Friess ◽  
SH Hartmann
Keyword(s):  
Mammary Gland ◽  
Basement Membrane ◽  
Immunoglobulin G ◽  
Passive Immunity ◽  
Morphological Evidence ◽  
Membrane Material ◽  
Ultrastructural Evidence ◽  
Cytotrophoblast Cells ◽  
Fetal Vessels

In dogs, passive immunity is conferred to fetuses and neonates by the transfer of maternal immunoglobulin G through the placenta during the last trimester of pregnancy and via the mammary gland after parturition, respectively. However, morphological evidence of transplacental transport is still lacking. The aim of the present study was to localize maternal immunoglobulin G in the labyrinthine zone and in the haemophagous zone of the canine placenta by means of immunohistochemistry and immunocytochemistry. In the labyrinthine zone, immunoglobulin G was detected in all the layers of the materno-fetal barrier including the fetal capillaries. Immunoreactivity was particularly prominent in maternal basement membrane material as well as in the syncytiotrophoblast. However, this evidence of transplacental transport of immunoglobulin G originated from a limited number of unevenly distributed maternal vessels only. In the cytotrophoblast of the haemophagous zone, immunoglobulin G was localized to phagolysosomes at various stages but was never detected within fetal vessels. The results indicate that maternal immunoglobulin G is degraded in cytotrophoblast cells of the hemophagous zone and, therefore, that transplacental transport is restricted to a subpopulation of maternal vessels in the labyrinthine zone.

Massive myoepithelial proliferation (myoepitheliosis) with lumpy deposits of basement membrane material closely associated with apocrine adenosis and ductal carcinomain situof the breast

2011 ◽  
Vol 61 (10) ◽  
pp. 615-617
Author(s):  
Tomonori Kawasaki ◽  
Toshio Oyama ◽  
Hiroshi Nakagomi ◽  
Kazushige Furuya ◽  
Tetsuo Kondo ◽  
...  
Keyword(s):  
Basement Membrane ◽  
Membrane Material

Spherulosis of the Breast

1999 ◽  
Vol 123 (7) ◽  
pp. 626-630 ◽  
Author(s):  
Eoghan E. Mooney ◽  
Naila Kayani ◽  
Fattaneh A. Tavassoli
Keyword(s):  
Basement Membrane ◽  
Invasive Carcinoma ◽  
Armed Forces ◽  
Membrane Material ◽  
Material Deposition ◽  
Immunohistochemical Profile

Abstract Objective.—Collagenous spherulosis of the breast is an uncommon localized pattern of basement membrane material deposition that may be mistaken for atypical proliferations or carcinoma. This report describes 9 cases in which the predominant or exclusive appearance of the spherules was basophilic instead of eosinophilic. Design.—The files of all cases of collagenous spherulosis diagnosed at the Armed Forces Institute of Pathology were reviewed to ascertain the frequency of diagnosis. Results.—Spherulosis with a predominantly basophilic pattern had a histochemical and immunohistochemical profile similar to collagenous spherulosis and was associated with more collagenous-appearing forms in 7 of 9 cases. Review of 81 cases showed that collagenous spherulosis was correctly diagnosed in 15% of referrals and was mistaken for intraductal or invasive carcinoma in 11% of cases. Conclusions.—Mucinous and collagenous patterns appear to be related forms of spherulosis. They are underrecognized by pathologists and maybe mistaken for atypia or malignancy.

Histochemical Differentiation of the Basement Membrane of the Mouse Seminiferous Tubule

1962 ◽  
Vol s3-103 (63) ◽  
pp. 385-391
Author(s):  
A. H. BAILLIE
Keyword(s):  
Basement Membrane ◽  
Protein Metabolism ◽  
Seminiferous Tubule ◽  
Glucuronic Acid ◽  
Chondroitin Sulphate ◽  
Hair Follicles ◽  
Ground Substance ◽  
Seminiferous Tubules ◽  
Active Protein ◽  
Pas Reaction

The ground substance of the testis of the albino mouse is PAS-positive but not metachromatic, and probably highly aggregated. The basement of the seminiferous tubules is intensely PAS-positive, metachromatic, and possibly not so highly aggregated. The reactivity of the ground substance to the PAS reaction and toluidine blue is tentatively ascribed to the presence of chondroitin sulphate C: this compound, previously known to contain N acetyl-galactosamine, glucuronic acid, tyrosine and tryptophane, is associated with arginine. The genesis of the basement membrane of the seminiferous tubule is shown to include the formation of a sheath of atypical elongated fibroblasts, the secretion of a PAS positive, metachromatic substance associated with arginine between this sheath and the seminiferous tubule, the appearance of mitochondria in the cells of the sheath, and lastly, the acquisition of alkaline phosphatase by these fibroblasts and its spread to the intervening ground substance. These changes are thought to be related to the structural and nutritional requirements of the seminiferous tubules. In its intense positive reaction to PAS and in its metachromasy, the basement membrane of the seminiferous tubule agrees with the ground substance adjacent to sites of active protein metabolism, such as growing tumours, embryonic organs, hair follicles, and skin.

Cell surface saccharides of Trypanosoma lewisi. I. Polycation-induced cell agglutination and fine-structure cytochemistry

1975 ◽  
Vol 19 (3) ◽  
pp. 621-644
Author(s):  
D.M. Dwyer
Keyword(s):  
Fine Structure ◽  
Cell Surface ◽  
Alcian Blue ◽  
Chondroitin Sulphate ◽  
Surface Membrane ◽  
Ruthenium Red ◽  
Surface Coat ◽  
Cell Agglutination ◽  
Trypanosoma Lewisi ◽  
Cationic Compounds

Trypanosoma lewisi bloodstream and culture forms were agglutinated differentially with low concentrations of the cationic compounds: ruthenium red, ruthenium violet, Alcian blue chloride, 1-hexadecylpyridinium chloride, lanthanum chloride, and cationized ferritin. The bloodstream form trypanosomes gave the highest agglutination levels with each of the compounds tested. Ruthenium red was the most effective inducer of cell agglutination among the several cations used. Trypsin-treated bloodstream forms were agglutinated less in the presence of ruthenium red than untreated controls. Ruthenium red-induced cell agglutination also was lowered with chondroitin sulphate and dextran sulphate, but not with alpha-D-glucose, alpha-D-mannose or with several methyl glycosides. Treatment of the bloodstream trypanosomes with alpha-amylase, dextranase, or neuraminidase had little effect on agglutination levels obtained with ruthenium red. Fine-structure cytochemical staining with ruthenium red, ruthenium violet, and Alcian blue-lanthanum nitrate was used to ascertain the presence and distribution of presumptive carbohydrates in the trypanosome cell surface. The extracellular surface coat of the bloodstream forms stained densely with each of the polycationic dyes. Trypsin treatment removed the surface coat from bloodstream trypanosomes; however, the surface membranes of the organisms were stained densely with the several dyes. Similar surface-membrane staining was obtained with the cationic compounds and the culture forms, which lack a cell surface coat. Cationized ferrin was used at the fine-structure level to visualize the negative surface charge present in the cell surface coat and external membrane of the several trypanosome stages. Results obrained from the agglutination and cytochemistry experiments indicate that complex polysaccharides are present in the surface membranes and cell surface coat of T. lewisi bloodstream forms. Similar conclusions also pertain to the surface membranes of the T. lewisi culture from trypanosomes. The carbohydrates probably represent glycopeptide and glycoprotein structural components of the surface membrane of this organism.

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