Preservation of intercellular substances by the cationic dye alcian blue in preparative procedures for electron microscopy

Behnke, O.; Zelander, T.

doi:10.1016/s0022-5320(70)90159-0

Cited by 268 publications

(100 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In both instances the size and distribution of these granules were similar, The granules were polygonal in shape, measured 200-500 A in diameter, and possessed weakly stained 30-60-A thick filamentous projections which appeared to interconnect adjacent granules (Figs. 4,5,6).…”

Section: Electron Microscopymentioning

confidence: 99%

“…In some regions, adjacent elastic fibers were interconnected by the matrix granules and their filaments (Figs. 4,6). The granules were often deposited on the surfaces of collagen fibrils at their major period…”

Section: Electron Microscopymentioning

confidence: 99%

“…Glycosaminoseveral connective tissues (1,4,9,18,23,26,28,31,33,38,41,42,46,47), relatively little is known concerning the ultrastructural distribution of these macromolecules in arteries.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Proteoglycans in primate arteries. I. Ultrastructural localization and distribution in the intima.

Wight¹,

Ross²

1975

The Journal of Cell Biology

184

View full text Add to dashboard Cite

Proteoglycans were identified and localized histochemically and ultrastructurally in normal and hyperplastic arterial intimas in nonhuman primates (Macaca nemestrina). These regions were consistently more alcianophilic than the adjacent medial layers and this alcianophilia was absent after treatment with glycosaminoglycan-degradative enzymes. Ultrastructurally, the intimal intercellular matrix consisted of numerous, irregularly shaped, 200-500-A diameter granules possessing 30-60-A diameter filamentous projections, and these granules were dispersed between collagen and elastic fibers. The granules exhibited a marked affinity for ruthenium red and were interconnected via their filamentous projections. The ruthenium red-positive granules were intimately associated with the plasma membrane of intimal smooth muscle cells and attached to collagen fibrils and elastic fibers. The matrix granules were completely removed after testicular hyaluronidase or chondroitinase ABC digestion but only partially removed after leech hyaluronidase treatment. These results suggest that the matrix granules contain some hyaluronic acid and one or more isomers of chondroitin sulfate. In addition to the large ruthenium red-positive matrix granules, a smaller class of ruthenium red-positive granule (100-200-A diameter) was present within the basement membranes beneath the endothelium and surrounding the smooth muscle cells. Ruthenium red also exhibited an affinity for the surface coat of the smooth muscle cells. The potential importance of proteoglycans in arterial intimal hyperplasia is discussed.Although the fine structural localization and distribution of proteoglycans 1 has been examined in ~Proteoglycans are defined as high molecular weight protein polysaccharides which consist of glycosaminoglycans (carbohydrate polymers composed of repeating dimers of monosaccharides, one of which is an amino sugar) covalently linked to a protein core. Glycosaminoseveral connective tissues (1,4,9,18,23,26,28,31,33,38,41,42,46,47), relatively little is known concerning the ultrastructural distribution of these macromolecules in arteries.glycans do not exist as free polysaccharides in the intercellular space but always occur bound to protein as a complex.

show abstract

Section: Electron Microscopymentioning

confidence: 99%

Section: Electron Microscopymentioning

confidence: 99%

See 1 more Smart Citation

Proteoglycans in primate arteries. I. Ultrastructural localization and distribution in the intima.

Wight¹,

Ross²

1975

The Journal of Cell Biology

184

View full text Add to dashboard Cite

show abstract

“…A number of studies has established that the polygonal matrix granules are composed, at least in part, of precipitated proteoglycan aggregates. These studies include the following criteria: Alcian blue stains localize the proteoglycan in these granules (2,5,30,34); prior treatment of cartilage with testicular hyaluronidase or chondroitinase ABC reduces the size and number of these matrix granules (18,34); extraction of cartilage with 4 M guanidine hydrochloride removes more than half 294 THE JOURNAL OF CELL BIOLOGY" VOLUME 73, 1977…”

Section: Discussionmentioning

confidence: 99%

Defects in the cartilaginous growth plates of brachymorphic mice.

Orkin¹,

Williams²,

Cranley³

et al. 1977

The Journal of Cell Biology

View full text Add to dashboard Cite

Homozygous brachymorphic (bm/bm) mice are characterized by disproportionately short stature. Newborn bm/bm epiphyseal cartilages are shorter than normal although the cells in the different zones of growth are relatively well organized. The extracellular matrix reacts poorly with stains specific for sulfated glycosaminoglycans. The ultrastructural appearance of the cartilage matrix indicates normal collagen fibrils; however, proteoglycan aggregate granules are smaller than normal and are present in reduced numbers, particularly in the columnar and hypertrophic zones of the growth plate. In addition, a prominent network of fine filaments, which are extractable in 4 M guanidine hydrochloride, are present in the bm/bm cartilage matrix. These findings suggest that a defect affecting the proteoglycan component of cartilage occurs in bm/bm mice.Numerous inherited disorders affect cartilage structure and function in man and other animals. Recently, we have studied cartilage from brachymorphic (bm/bm) mice. This mutant was first described by Lane and Dickie (12), and established by genetic studies as an autosomal recessive condition. Homozygous bm/brn mice are vigorous but have shortened long bones, shortened tails, and dome-shaped skulls. Such findings are consistent with a defect in cartilage development, and Lane and Dickie found the knee joints of 17-24 day old bin~bin mice to be shorter and thicker than normal, although chondrocytes in these epiphyseal growth plates retained good alignment (12).Cartilage is composed predominantly of one cell type, chondrocytes, surrounded by large quantities of extracellular matrix. Of the constituents of the extracellular matrix, two macromolecules, cartilage-type collagen (19, 36) and cartilage-specific proteoglycans (13,22), are synthesized by the differentiated chondrocytes. Ultrastructurally, this extracellular matrix consists of thin, 200-300 ,~ wide cartilage-type collagen fibrils and polygonal matrix granules of precipitated proteoglycan aggregates (1,6,18,25,(33)(34)(35).In examining the cartilaginous epiphyses from bm/bm mice, we have observed alterations in the morphological appearance of the epiphyseal growth plates at different stages of development, as well as differences in the extracellular matrix material which may be related to the abnormalities in tissue development found in this mutant.

show abstract

“…Thick sections (2 to 3 pm) were stained with toluidine blue (0.05 %, w/v, in 0.1 M-phosphate buffer pH 7-2). Polysaccharides were located using the Alcian blue staining procedure of Behnke & Zelander (1970).…”

Section: Introductionmentioning

confidence: 99%

Effect of Oxygen Concentration, Temperature and Combined Nitrogen on the Morphology and Nitrogenase Activity of Rhizobium sp. Strain 32H1 in Agar Culture

Pankhurst¹,

Craig²

1978

Journal of General Microbiology

View full text Add to dashboard Cite

207Cultures of Rhizobium sp. strain 3 2~1 grown in a layer of soft agar on top of a layer of harder agar developed maximum nitrogenase activity [80 to 90 nmol acetylene reduced h-l (mg protein)-l] after 12 d in air at 28 "C. Electron microscopy of sections of cores of the soft agar layer showed differences in the morphology of rhizobia in colonies growing at the surface, in the middle or at the bottom of the layer. The properties of rhizobia growing in the colonies in the middle of the soft agar layer suggested that only these contained nitrogenase. These rhizobia were present as a distinct band of cells passing through individual colonies at a constant depth in the agar. They were larger than normal vegetative Rhizobium, were pleiomorphic and were similar in morphology to the nitrogen-fixing bacteroids formed by strain 3 2~1 in cowpea root nodules. Their location within the soft agar layer changed with the O2 concentration in the atmosphere above the agar during growth, and they were not found in cultures showing little or no nitrogenase activity. Loss of nitrogenase activity occurred when cultures were grown at 33 "C, in a 02/Ar (1 :4, v/v) atmosphere, or when the concentration of combined nitrogen in the agar medium was increased from 2 to 20 mM-N.

show abstract

Preservation of intercellular substances by the cationic dye alcian blue in preparative procedures for electron microscopy

Cited by 268 publications

References 15 publications

Proteoglycans in primate arteries. I. Ultrastructural localization and distribution in the intima.

Proteoglycans in primate arteries. I. Ultrastructural localization and distribution in the intima.

Defects in the cartilaginous growth plates of brachymorphic mice.

Effect of Oxygen Concentration, Temperature and Combined Nitrogen on the Morphology and Nitrogenase Activity of Rhizobium sp. Strain 32H1 in Agar Culture

Contact Info

Product

Resources

About