SPECIFIC STAINING OF SULPHATE GROUPS WITH ALCIAN BLUE AT LOW pH

Lev, Robert; Spicer, S. S.

doi:10.1177/12.4.309

Cited by 841 publications

(314 citation statements)

References 0 publications

Supporting

Mentioning

298

Contrasting

Unclassified

Order By: Relevance

“…Saccharide moieties were investigated on denaturing mini-gels by use of periodic acid Schiff (PAS) [63] and Alcian Blue 8GX staining, [64] which at pH 1 is specific for sulfated sugars. [65] The calcium-binding ability of the ASM was investigated by "Stains-All" (cationic carbocyanine dye) staining [66] and by the 45 Ca overlay test. [67] For the second of these, the gel was transferred to a PVDF Immobilon-P membrane (Millipore Corp.), with a mini-Trans Blot module (Bio-Rad) for 90 min at 120 mA.…”

Section: Methodsmentioning

confidence: 99%

Evolution of Nacre: Biochemistry and Proteomics of the Shell Organic Matrix of the Cephalopod Nautilus macromphalus

Marie¹,

Marin²,

Marie³

et al. 2009

ChemBioChem

View full text Add to dashboard Cite

Matrix evolutions: We have biochemically characterized the nacre matrix of the cephalopod Nautilus macromphalus, in part by a proteomic approach applied to the acetic acid‐soluble and ‐insoluble shell matrices, as well as to spots obtained after 2D gel electrophoresis. Strikingly, most of the obtained partial sequences are entirely new, whereas a few correspond only partly with bivalvian nacre proteins. Our findings shed new light on the macroevolution of nacre matrix proteins. magnified imageIn mollusks, one of the most widely studied shell textures is nacre, the lustrous aragonitic layer that constitutes the internal components of the shells of several bivalves, a few gastropods, and one cephalopod: the nautilus. Nacre contains a minor organic fraction, which displays a wide range of functions in relation to the biomineralization process. Here, we have biochemically characterized the nacre matrix of the cephalopod Nautilus macromphalus. The acid‐soluble matrix contains a mixture of polydisperse and discrete proteins and glycoproteins, which interact with the formation of calcite crystals. In addition, a few bind calcium ions. Furthermore, we have used a proteomic approach, which was applied to the acetic acid‐soluble and ‐insoluble shell matrices, as well as to spots obtained after 2D gel electrophoresis. Our data demonstrate that the insoluble and soluble matrices, although different in their bulk monosaccharide and amino acid compositions, contain numerous shared peptides. Strikingly, most of the obtained partial sequences are entirely new. A few only partly match with bivalvian nacre proteins. Our findings have implications for knowledge of the long‐term evolution of molluskan nacre matrices.

show abstract

Section: Methodsmentioning

confidence: 99%

Evolution of Nacre: Biochemistry and Proteomics of the Shell Organic Matrix of the Cephalopod Nautilus macromphalus

Marie¹,

Marin²,

Marie³

et al. 2009

ChemBioChem

View full text Add to dashboard Cite

show abstract

“…24 Briefly, tissues were deparaffinized and hydrated in distilled water, stained in alcian blue solution for 30 min and washed in running water for 10 min. Specimens were further stained in Schiff's reagent, rinsed in sodium metabisulfite, washed briefly in running water, dehydrated in ascending grades of alcohol, cleared in xylene and mounted with Pertex (Histolab, Göteborg, Sweden).…”

Section: Mucin Stainingmentioning

confidence: 99%

Downregulation of estrogen‐metabolizing 17β‐hydroxysteroid dehydrogenase Type 2 expression correlates inversely with Ki67 proliferation marker in colon‐cancer development

Oduwole

Isomaa

Nokelainen

et al. 2001

Intl Journal of Cancer

View full text Add to dashboard Cite

The 17HSDs are a group of isozymes that catalyze the interconversion between high-activity 17␤-hydroxysteroids and low-activity 17-ketosteroids. In the present study, we characterized the expression of 17HSD types 1 and 2 in normal and malignant gastrointestinal tissues and cells. Using the colon as a model for cancer of the gastrointestinal tract, expression of the 17HSD enzymes in cancer development was studied and correlated with proliferation and differentiation markers as assessed by Ki67 and mucin staining, respectively. In normal colon and small intestine, 17HSD type 2 mRNA was expressed in the surface epithelial cells and, to a lesser extent, in the cryptal epithelial cells. In colon-cancer specimens, 17HSD type 2 expression was downregulated both in the tissues and in the cell lines and correlated inversely with the proliferation marker. No expression for the 17HSD type 1 enzyme was observed in normal or cancerous gastrointestinal tract tissues. In line with the expression studies, 17HSD activity measurements with colon cells showed that only the oxidative conversion of E2 to E1 was present, and Northern blot analysis showed the signal only for 17HSD type 2. Localization of the ERs ␣ and ␤, assessed by immunohistochemistry and in situ hybridization, showed the presence of ER␤ in the lamina propria of the colon. Our study shows that 17HSD type 2 expression is associated with the functional integrity of the gastrointestinal tract. The decrease in expression of the type 2 enzyme may increase estrogen influence in colon cancer. © 2002 Wiley-Liss, Inc. Key words: colon carcinoma; estrogen; hydroxysteroid dehydrogenase; cell proliferation; cell differentiationEpidemiologic studies 1,2 and in vitro studies with colon-cancer cell lines 3 suggest the involvement of estrogens in the development and progression of gastrointestinal cancer. [3][4][5] Contrary to this, however, is the indication that HRT is effective at reducing the incidence of colon cancer. 6 The mechanism underlying the sex steroid-mediated effects in normal gastrointestinal physiology and its involvement in neoplastic development and progression, if any, are far from clear.The 17HSDs play pivotal roles in the regulation of the physiologic activities of sex steroid hormones by catalyzing the oxidation or reduction of 17␤-hydroxy-and 17-ketosteroids, respectively. 7,8 17HSDs regulate the concentrations of the active sex steroids that bind to the respective steroid receptors to regulate gene expression in target cells. To date, at least 8 isoforms of the 17HSD enzyme with different substrate and cofactor specificities, tissue distributions and subcellular localizations have been characterized. 8,9 Of these isoforms, 17HSD type 1 and type 2 catalyze opposite reactions of estrogen metabolism. 17HSD type 1 is the predominant enzyme involved in estradiol biosynthesis (E1 to E2), and it is expressed in cells of placental and ovarian origin 10,11 as well as in breast tissue. 12 On the other hand, 17HSD type 2 catalyzes the oxidation of estrogens (E2 to E1...

show abstract

“…Briefly, net absorbance a t 600 nm (A6,,) per well was measured in 4 M guanidinium HC1 (Sigma) extracts of Alcian-blue (ICN, Cleveland, OH) stained cartilage matrix from fixed 4-day cultures. Alcian blue binds quantitatively to sulfated glycosaminoglycans (Lev and Spicer, 1964). Bound and extracted stain in micromass cultures of limb-bud mesenchyme is proportionate to accumulated sulfated proteoglycan (Biddulph et al, 1988;Leonard et al, 1989).…”

Section: Chondrogenesismentioning

confidence: 99%

Stage‐ and region‐dependent chondrogenesis and growth of chick wing‐bud mesenchyme in serum‐containing and defined tissue culture media

Paulsen

Chen

Pang

et al. 1994

Developmental Dynamics

View full text Add to dashboard Cite

During development, limb-bud mesenchymal cells carry out complex spatiotemporal patterns of growth and differentiation. Tissue and organ culture facilitate analysis of environmental influences on these cell behaviors, allowing their partial dissection into exogenous and endogenous components. Two factors that complicate such in vitro analyses are the heterogeneity of the cultured cells and imprecise knowledge of culture medium composition. Limb mesenchyme comprises a heterogenous cell population with important regional differences in cell type. Dividing the limb into subregions helps limit the cellular heterogeneity and using chemically defined, serum-free medium allays concerns about medium composition. In the present study, mesenchyme from different regions along the anteroposterior and proximodistal axes of stage 21-22 and stage 23-24 chick wing buds was grown in high-density microtiter cultures in chemically defined and in serum-containing medium. Fourday cultures of the various regions were compared in terms of culture morphology and the accumulation of Alcian blue-positive cartilage matrix and DNA. The results demonstrate stageand region-dependent differences in the in vitro growth, differentiation, and responsiveness of these cells. For example, mesenchyme from the distal anterior region of the wing bud exhibited lower intrinsic chondrogenic capacity and greater responsiveness to serum than other regions. Patterns of in vitro chondrogenesis also suggest that, at the stages examined, distal wing-bud mesenchyme may be less homogeneous than has been believed. A case is made for the suitability of serum-free medium for future in vitro studies of chick limb-bud mesenchyme. The results are considered in relation to the process of limb development and regional expression of pattern-related genes. 0 1994 Wiley-Liss, Inc.

show abstract

SPECIFIC STAINING OF SULPHATE GROUPS WITH ALCIAN BLUE AT LOW pH

Cited by 841 publications

References 0 publications

Evolution of Nacre: Biochemistry and Proteomics of the Shell Organic Matrix of the Cephalopod Nautilus macromphalus

Evolution of Nacre: Biochemistry and Proteomics of the Shell Organic Matrix of the Cephalopod Nautilus macromphalus

Downregulation of estrogen‐metabolizing 17β‐hydroxysteroid dehydrogenase Type 2 expression correlates inversely with Ki67 proliferation marker in colon‐cancer development

Stage‐ and region‐dependent chondrogenesis and growth of chick wing‐bud mesenchyme in serum‐containing and defined tissue culture media

Contact Info

Product

Resources

About