Immunocytology on microwave-fixed cells reveals rapid and agonist-specific changes in subcellular accumulation patterns for cAMP or cGMP.

Bársony, Julia; Marx, Stephen J.

doi:10.1073/pnas.87.3.1188

Cited by 67 publications

(25 citation statements)

References 25 publications

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“…This poor correlation is probably caused by determination of overall cellular cAMP content. Dependent on intracellular PDE distribution and on mode of adenylate cyclase activation high local subcellular cAMP peaks are likely to occur, as has already been demonstrated in microwave-fixed cells (37). These cAMP peaks probably are easily missed by global cAMP determinations in cell lysates.…”

Section: Resultsmentioning

confidence: 84%

Role of phosphodiesterases in the regulation of endothelial permeability in vitro.

Suttorp¹,

Weber²,

Welsch³

et al. 1993

J. Clin. Invest.

176

143

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Neutrophil-derived hydrogen peroxide (H202) is believed to play an important role in the pathogenesis of vascular injury and pulmonary edema. H202 time-and dose-dependently increased the hydraulic conductivity and decreased the selectivity of an endothelial cell monolayer derived from porcine pulmonary arteries. Effects of H202 on endothelial permeability were completely inhibited by adenylate cyclase activation with 10-12 M cholera toxin or 0.1 juM forskolin. 10' M Sp-cAMPS, a cAMP-dependent protein kinase A agonist, was similarly effective. The phosphodiesterase (PDE) inhibitors motapizone (10'-M), rolipram (10-' M), and zardaverine (10-8 M), which specifically inhibit PDE-isoenzymes III, IV, and III/IV potently blocked H202-induced endothelial permeability when combined with 10' M prostaglandin El. Overall cellular cAMP content and inhibition of H202 effects on endothelial permeability were poorly correlated. H202 exposure resulted in a rapid and substantial decrease in endothelial cAMP content. The analysis of the PDE isoenzyme spectrum showed high activities of isoenzymes II, III, and IV in porcine pulmonary endothelial cells. The data suggest that adenylate cyclase activation/PDE inhibition is a powerful approach to block H202-induced increase in endothelial permeability. This concept appears especially valuable when endothelial PDE isoenzyme pattern and PDE inhibitor profile are matched optimally. (J.

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Section: Resultsmentioning

confidence: 84%

Role of phosphodiesterases in the regulation of endothelial permeability in vitro.

Suttorp¹,

Weber²,

Welsch³

et al. 1993

J. Clin. Invest.

176

143

View full text Add to dashboard Cite

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“…The kinetics of the accumulation of and differences in the intracellular localization of cAMP may need to be considered. In this context Barsony and Marx (1990) have shown, using immunohistochemical techniques, that the site of accumulation of cAMP varies depending upon the agonist used to stimulate cAMP production. Forskolin, for example, can cause nuclear accumulation of cAMP whereas calcitonin causes mainly cytoplasmic and perinuclear accumulation.…”

Section: Discussionmentioning

confidence: 99%

Interaction between cAMP elevation, identified growth factors, and serum components in regulating Schwann cell growth

Stewart

Eccleston

Jessen

et al. 1991

J of Neuroscience Research

109

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Most previous studies on Schwann cell proliferation in vitro have used serum-containing media. This complicates the analysis of agents required for cell division since serum contains an ill-defined mixture of hormones and growth factors. Serum-free medium has therefore been used to analyse the response of Schwann cell to previously identified Schwann cell mitogens. Serum factors were not necessary for DNA synthesis in response to platelet-derived growth factor, basic fibroblast growth factor, or glial growth factor, provided they were used in combination with forskolin to elevate intracellular cAMP. Transforming growth factor beta 1, a Schwann cell mitogen in serum, was not mitogenic under these conditions. Neither the growth factors nor forskolin were effective when used alone. Growth control was analysed further using long-term cultured Schwann cells that had spontaneously immortalized. Measurements of endogenous cAMP levels in short- and long-term Schwann cells revealed that long-term cells had two to three times higher basal cAMP levels. As predicted by these findings, platelet-derived growth factor, basic fibroblast growth factor, and glial growth factor stimulated DNA synthesis in long-term cells without requiring costimulation by agents which elevate cAMP (while transforming growth factor beta 1 had no effect).(ABSTRACT TRUNCATED AT 250 WORDS)

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“…T welve-micrometer-thick sections were cut and stored at Ϫ70°C until use. Immediately before immunostaining, the sections were microwave-irradiated in the presence of 2% paraformaldehyde (Barsony and Marx, 1990). Fixed sections were blocked with 3% normal horse serum in PBS and then incubated with RB18 anti-brevican hybridoma culture supernatant at 1:5 dilution for 1 hr.…”

Section: Methodsmentioning

confidence: 99%

The Brain Chondroitin Sulfate Proteoglycan Brevican Associates with Astrocytes Ensheathing Cerebellar Glomeruli and Inhibits Neurite Outgrowth from Granule Neurons

et al. 1997

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Brevican is a nervous system-specific chondroitin sulfate proteoglycan that belongs to the aggrecan family and is one of the most abundant chondroitin sulfate proteoglycans in adult brain. To gain insights into the role of brevican in brain development, we investigated its spatiotemporal expression, cell surface binding, and effects on neurite outgrowth, using rat cerebellar cortex as a model system. Immunoreactivity of brevican occurs predominantly in the protoplasmic islet in the internal granular layer after the third postnatal week. Immunoelectron microscopy revealed that brevican is localized in close association with the surface of astrocytes that form neuroglial sheaths of cerebellar glomeruli where incoming mossy fibers interact with dendrites and axons from resident neurons. In situ hybridization showed that brevican is synthesized by these astrocytes themselves. In primary cultures of cerebellar astrocytes, brevican is detected on the surface of these cells. Binding assays with exogenously added brevican revealed that primary astrocytes and several immortalized neural cell lines have cell surface binding sites for brevican core protein. These cell surface brevican binding sites recognize the C-terminal portion of the core protein and are independent of cell surface hyaluronan. These results indicate that brevican is synthesized by astrocytes and retained on their surface by an interaction involving its core protein. Purified brevican inhibits neurite outgrowth from cerebellar granule neurons in vitro, an activity that requires chondroitin sulfate chains. We suggest that brevican presented on the surface of neuroglial sheaths may be controlling the infiltration of axons and dendrites into maturing glomeruli.

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Immunocytology on microwave-fixed cells reveals rapid and agonist-specific changes in subcellular accumulation patterns for cAMP or cGMP.

Cited by 67 publications

References 25 publications

Role of phosphodiesterases in the regulation of endothelial permeability in vitro.

Role of phosphodiesterases in the regulation of endothelial permeability in vitro.

Interaction between cAMP elevation, identified growth factors, and serum components in regulating Schwann cell growth

The Brain Chondroitin Sulfate Proteoglycan Brevican Associates with Astrocytes Ensheathing Cerebellar Glomeruli and Inhibits Neurite Outgrowth from Granule Neurons

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