Hyaluronan Oligosaccharides Induce Tube Formation of a Brain Endothelial Cell Linein Vitro☆

Rahmanian, Mehdi; Pertoft, Håkan; Kanda, Shigeru; Christofferson, Rolf; Claesson‐Welsh, Lena; Heldin, Paraskevi

doi:10.1006/excr.1997.3792

Cited by 62 publications

(48 citation statements)

References 20 publications

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“…5,13 We also observed that hyaluronan dodecasaccharides induced endothelial cells grown in a 3-dimensional (3D) collagen gel to form tubular structures, in a process not mediated through activation of FGF receptors. 14 These results are in conformity with those of Montesano and his colleagues, 15 who found that hyaluronan fragments acted synergistically with VEGF, but not with bFGF, in the induction of endothelial cell tube formation. The cellular effects of hyaluronan and hyaluronan oligosaccharides are mediated through interactions with hyaluronan-binding proteins, the hyaladherins.…”

supporting

confidence: 91%

“…5,13,14 An important question is whether hyaluronidase induces endothelial cell tube formation through the formation of hyaluronan fragments with angiogenic activity or through another yet unknown mechanism. If the mechanism involves production of hyaluronan fragments, hyaluronan needs to be present in the assay.…”

Section: Discussionmentioning

confidence: 99%

“…14 Briefly, 8/10 volumes of type I collagen solution, 1/10 volume of 10 ϫ HamЈs F-12 medium, containing 4 mM Lglutamine and 1/10 volume of 260 mM NaHCO 3 , 200 mM HEPES and 50 mM NaOH were mixed on ice. Then aliquots of 500 l/well were poured into 12-well Falcon culture plates and allowed to gel at 37°C for 1 hr.…”

Section: In Vitro Angiogenesis Assaymentioning

confidence: 99%

“…14 In an attempt to elucidate whether CD44, which is implicated in hyaluronan uptake and degradation, also participates in hyaluronidase-induced tube formation of endothelial cells, we investigated whether KM114, a monoclonal antibody that blocks the binding of hyaluronan to CD44 receptors, 39 had any role in tube formation. KM114 antibodies at a concentration of 40 g/ml blocked the binding of 3 H-hyaluronan to CD44 receptors by about 65%, suggesting that CD44 is the predominant hyaluronan receptor expressed on the surface of endothelial cells (data not shown).…”

Section: Blocking Of Cd44 Receptors Prevents Hyaluronidase-induced Tumentioning

confidence: 99%

See 3 more Smart Citations

Testicular hyaluronidase induces tubular structures of endothelial cells grown in three‐dimensional colagen gel through a CD44‐mediated mechanism

Rahmanian

Heldin

2001

Intl Journal of Cancer

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Cultured brain capillary endothelial cells grown in a 3-dimensional collagen gel can form tubular structures after stimulation by angiogenic factors. We found that treatment of such cultures with testicular hyaluronidase led to formation of tubular structures and cell survival. Anion-exchange chromatography of the enzyme preparation on a MonoQ column revealed the presence of the angiogenic factor basic fibroblast growth factor (bFGF) in the flow-through fraction, as determined by immunoblotting; part of the effect on endothelial cell morphogenesis could thus be ascribed to bFGF. However, adsorbed fractions eluted with increasing concentrations of NaCl, which exhibited hyaluronan-degrading activity at neutral pH, did not contain bFGF but were still able to induce tube-like structures of the endothelial cells. Streptomyces hyaluronidase failed to evoke the same effect. Interestingly, blocking of hyaluronan binding to CD44 receptors by the monoclonal antibody KM114 inhibited the effect of hyaluronidase, but not of bFGF, on endothelial cell tube formation. Our data suggest a CD44-mediated specific role for certain populations of testicular hyaluronidase in the induction of angiogenesis.

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supporting

confidence: 91%

Section: Discussionmentioning

confidence: 99%

Section: In Vitro Angiogenesis Assaymentioning

confidence: 99%

Section: Blocking Of Cd44 Receptors Prevents Hyaluronidase-induced Tumentioning

confidence: 99%

See 2 more Smart Citations

Testicular hyaluronidase induces tubular structures of endothelial cells grown in three‐dimensional colagen gel through a CD44‐mediated mechanism

Rahmanian

Heldin

2001

Intl Journal of Cancer

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“…For example, low molecular weight HA fragments stimulate endothelial cells to form tube-like structures reminiscent of angiogenic vessels. By contrast, high molecular weight HA does not stimulate endothelial tube formation [5].…”

Section: Introductionmentioning

confidence: 91%

Development of a fluorescent substrate to measure hyaluronidase activity

Zhang

Mummert

2008

Analytical Biochemistry

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A novel fluorescent substrate (termed FRET-HA) to quantitatively assess hyaluronidase activity was developed. Hyaluronan (HA), the major substrate for hyaluronidase, was dual labeled with fluorescein amine and rhodamine B amine. The fluorescein amine fluorescence signal was significantly quenched while the rhodamine B amine signal was significantly enhanced due to fluorescence resonance energy transfer (FRET). In the presence of bovine testes hyaluronidase, cleavage of HA disrupted FRET resulting in a loss of the fluorescein amine quenching that was dependent upon both enzyme concentration and time. Increase in the fluorescein amine signal could be conveniently monitored in both non-continuous and continuous fashions. The K m value for bovine testes hyaluronidase was determined using FRET-HA in a continuous fluorescent assay. Importantly, the estimated K m value for bovine testes hyaluronidase using FRET-HA as the substrate was in excellent agreement with K m values previously reported for this enzyme using native (i.e., unlabeled) HA. Therefore, FRET-HA is a reliable substrate for quantitatively assessing the HA/ hyaluronidase molecular interaction. The simplicity, sensitivity and versatility of the FRET-HA substrate suggest that it will have utility in a variety of assay platforms and should be a new tool for assessing hyaluronidase activity.

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Molecular cloning and characterization of antigens expressed on rat tumor vascular endothelial cells

Taniguchi¹,

Harada²,

Ohizumi³

et al. 2000

Int. J. Cancer

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Hyaluronan Oligosaccharides Induce Tube Formation of a Brain Endothelial Cell Linein Vitro☆

Cited by 62 publications

References 20 publications

Testicular hyaluronidase induces tubular structures of endothelial cells grown in three‐dimensional colagen gel through a CD44‐mediated mechanism

Testicular hyaluronidase induces tubular structures of endothelial cells grown in three‐dimensional colagen gel through a CD44‐mediated mechanism

Development of a fluorescent substrate to measure hyaluronidase activity

Molecular cloning and characterization of antigens expressed on rat tumor vascular endothelial cells

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