Effects of beta-xylosides on proteoglycan biosynthesis and morphology of PC12 pheochromocytoma cells and primary cultures of rat cerebellum

1991 ◽  
Vol 99 (2) ◽  
pp. 237-246
Author(s):  
R.K. Margolis ◽  
B. Goossen ◽  
H. Tekotte ◽  
L. Hilgenberg ◽  
R.U. Margolis
Keyword(s):  
Heparan Sulfate ◽  
Chondroitin Sulfate ◽  
Pc12 Cells ◽  
Primary Cultures ◽  
Fold Increase ◽  
Pheochromocytoma Cells ◽  
Rat Cerebellum ◽  
Explant Cultures ◽  
Collagen Substratum ◽  
Pc12 Pheochromocytoma Cells

We have examined the effects of beta-xylosides, which act as exogenous acceptors for glycosaminoglycan chain initiation, on the morphology and proteoglycan biosynthesis of PC12 pheochromocytoma cells, and on monolayer, aggregate and explant cultures of early postnatal rat cerebellum. PC12 cells cultured for 13 days in the presence of nerve growth factor (NGF) and beta-xyloside, and labeled during days 11–13 with sodium [35S]sulfate, showed an 8- to 11-fold increase in [35S]sulfate-labeled macromolecules released into the culture medium. Most of the increase was accounted for by chondroitin sulfate, which was in the form of free glycosaminoglycan chains, which were not acid-precipitable. The presence of beta-xyloside also led to a 65–115% increase in [35S]sulfate incorporation into cell-associated glycosaminoglycans and glycoproteins of untreated and NGF-treated PC12 cells, respectively. beta-Xyloside treatment reduced the size of the chondroitin sulfate chains in both the cells and medium from approximately 34,000 to 10,000 Mr, but had much less effect on heparan sulfate, which decreased in size from 16,000 to 13,000-14,500 Mr (in the medium and cells, respectively). beta-Xyloside inhibition of proteoglycan biosynthesis was accompanied by significant morphological effects in NGF-treated PC12 cells, consisting of an increase in length and decrease in the branching, diameter and adhesion to the collagen substratum of the PC12 cell processes. p-Nitrophenyl- and 4-methylumbelliferyl-beta-D-xylosides produced similar effects, which were not seen with p-nitrophenyl-beta-D-galactoside. beta-Xylosides also produced distinct alterations in the adhesion and morphology of monolayer, aggregate, and explant cultures of early postnatal rat cerebellum, which occurred together with inhibition of chondroitin sulfate proteoglycan biosynthesis and a decrease in glycosaminoglycan chain size. These studies indicate that chondroitin sulfate (and probably also heparan sulfate) proteoglycans play a significant role in modulating cell-cell and cell-matrix interactions in nervous tissue development and differentiation.

Antiproliferative effect of suramin on primary cultures of human pheochromocytomas and rat PC12 pheochromocytoma cells

1997 ◽  
Vol 66 (1) ◽  
pp. 11-18 ◽  
Author(s):  
Andreas Zielke ◽  
Mariwil G. Wong ◽  
Allan E. Siperstein ◽  
Orlo H. Clark ◽  
Matthias Rothmund ◽  
...  
Keyword(s):  
Primary Cultures ◽  
Antiproliferative Effect ◽  
Pheochromocytoma Cells ◽  
Pc12 Pheochromocytoma Cells

scholarly journals pG2 gene expression and its regulation in human adrenocortical and medullary tumors

1999 ◽  
pp. 590-596
Author(s):  
J Liu ◽  
R Voutilainen ◽  
P Heikkila ◽  
AI Kahri
Keyword(s):  
Kinase Inhibitor ◽  
Primary Cultures ◽  
Control Level ◽  
Fold Increase ◽  
Zona Glomerulosa ◽  
Adrenal Tumors ◽  
Specific Marker ◽  
Mrna Accumulation ◽  
Adrenal Cells ◽  
Pheochromocytoma Cells

The cDNA clone pG2 was originally isolated from a human pheochromocytoma. The respective gene was found to be strongly expressed in normal adrenal zona glomerulosa and medulla, as well as in Conn's adenomas and pheochromocytomas. To shed more light on the expression and regulation of the pG2 gene, we investigated its expression in a wide variety of different adrenal neoplasms and cultured adrenal cells. Northern blot analysis was used to determine the steady state level of pG2 mRNA. Besides normal adrenals, Conn's adenomas and pheochromocytomas, we found abundant expression of pG2 mRNA in Cushing's, virilizing and nonfunctional adrenocortical adenomas and carcinomas, as well as in hyperplastic adrenals. The relative levels of pG2 mRNA in various adrenocortical tumors were not significantly different from those in normal adrenals and pheochromocytomas. In primary cultures of normal adrenal cells, treatment with adrenocorticotropin induced a 3- to 15-fold increase in the expression of pG2 mRNA (P<0.01), and this effect was reproduced by incubation with (Bu)2cAMP. In cultured pheochromocytoma cells, treatment with (Bu)2cAMP and a protein kinase inhibitor, staurosporine, increased pG2 mRNA accumulation (2- to 4-fold over the control level, P<0.01, and 3- to 8-fold, P<0.01, respectively). These results indicate that pG2 is widely expressed in normal and pathological adrenal tissues from both cortical and medullary origin, which eliminates its usefulness as a specific marker for zona glomerulosa or medullary adrenal tumors. Accumulation of pG2 mRNA is regulated by multiple differentiating factors through different pathways in primary cultures of normal adrenal and pheochromocytoma cells.

scholarly journals Synthesis by Schwann cells of basal lamina and membrane-associated heparan sulfate proteoglycans.

1985 ◽  
Vol 101 (2) ◽  
pp. 660-666 ◽  
Author(s):  
H Mehta ◽  
C Orphe ◽  
M S Todd ◽  
C J Cornbrooks ◽  
D J Carey
Keyword(s):  
Heparan Sulfate ◽  
Chondroitin Sulfate ◽  
Schwann Cells ◽  
Basal Lamina ◽  
Guanidine Hydrochloride ◽  
Primary Cultures ◽  
Sensory Nerve ◽  
Low Carbohydrate ◽  
Schwann Cell Development ◽  
Triton X

Primary cultures that contain only Schwann cells and sensory nerve cells synthesize basal lamina. The assembly of this basal lamina appears to be essential for normal Schwann cell development. In this study, we demonstrate that Schwann cells synthesize two major heparan sulfate-containing proteoglycans. Both proteoglycans band in dissociative CsCl gradients at densities less than 1.4 g/ml, and therefore, presumably, have relatively low carbohydrate-to-protein ratios. The larger of these proteoglycans elutes from Sepharose CL-4B in 4 M guanidine hydrochloride (GuHCl) at a Kav of 0.21 and contains heparan sulfate and chondroitin sulfate chains of Mr 21,000 in a ratio of approximately 3:1. This proteoglycan is extracted from cultures by 4 M GuHCl but not Triton X-100 and accumulates only when Schwann cells are actively synthesizing basal lamina. The smaller proteoglycan elutes from Sepharose CL-4B at a Kav of 0.44 and contains heparan sulfate and chondroitin sulfate chains of Mr 18,000 in a ratio of approximately 4:1. This proteoglycan is extracted by 4 M GuHCl or by Triton X-100. The accumulation of this proteoglycan is independent of basal lamina production.

Nerve growth factor (NGF)-dependent kinase activities in PC12 pheochromocytoma cells

1988 ◽  
Vol 117 (4_Suppl) ◽  
pp. S51
Author(s):  
ANKE-PEGGY HOLTORF ◽  
K. UNSICKER ◽  
H.-D. HOFMANN
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Nerve Growth ◽  
Pheochromocytoma Cells ◽  
Pc12 Pheochromocytoma Cells
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