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Article ; Online: Potential Influence of Endothelial Adsorption on the Delayed Time to Maximum Concentration of Biopharmaceuticals.

Reijers, Joannes A A / Dane, Martijn J C / van Zonneveld, Anton Jan / Burggraaf, Jacobus / Moerland, Matthijs

European journal of drug metabolism and pharmacokinetics

2018 Volume 43, Issue 1, Page(s) 103–113

Abstract: Background and objectives: Maximum plasma concentration of biopharmaceuticals sometimes occurs long after completion of intravenous infusion. The objective of this research was to study the hypothetical adsorption of biopharmaceuticals to endothelium ... ...

Abstract	Background and objectives: Maximum plasma concentration of biopharmaceuticals sometimes occurs long after completion of intravenous infusion. The objective of this research was to study the hypothetical adsorption of biopharmaceuticals to endothelium and infusion material, which may theoretically explain this phenomenon. Methods: Infusion procedures were mimicked in an artificial vessel covered with a confluent monolayer of endothelial cells. Three monoclonal antibodies (MAbs) and C1 inhibitor were studied. Results: Adsorption of MAbs to endothelium was observed followed by release when the vessel was subsequently perfused with buffer. Adsorption to infusion material also occurred to various degrees and in a seemingly random fashion, with a loss of up to 15% during a single flush of the line, but release from the line was not seen. Conclusions: Our results indicate that adsorption of biopharmaceuticals to endothelium can occur. This observation can explain the increase in plasma concentration after completion of intravenous administration.
MeSH term(s)	Adsorption ; Antibodies, Monoclonal/pharmacokinetics ; Bevacizumab/pharmacokinetics ; Biopharmaceutics ; Cells, Cultured ; Complement C1 Inhibitor Protein/administration & dosage ; Complement C1 Inhibitor Protein/pharmacokinetics ; Endothelium/metabolism ; Humans ; Immunoglobulin G/administration & dosage ; Immunoglobulin G/metabolism ; Infusions, Intravenous ; Time Factors ; Trastuzumab/pharmacokinetics
Chemical Substances	Antibodies, Monoclonal ; Complement C1 Inhibitor Protein ; Immunoglobulin G ; Bevacizumab (2S9ZZM9Q9V) ; Trastuzumab (P188ANX8CK)
Language	English
Publishing date	2018-02
Publishing country	France
Document type	Journal Article
ZDB-ID	196729-0
ISSN	2107-0180 ; 0398-7639 ; 0378-7966
ISSN (online)	2107-0180
ISSN	0398-7639 ; 0378-7966
DOI	10.1007/s13318-017-0430-1
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Zs.A 1236: Show issues

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Article ; Online: The endothelial glycocalyx as a potential modifier of the hemolytic uremic syndrome.

Boels, Margien G S / Lee, Dae Hyun / van den Berg, Bernard M / Dane, Martijn J C / van der Vlag, Johan / Rabelink, Ton J

European journal of internal medicine

2013 Volume 24, Issue 6, Page(s) 503–509

Abstract: Atypical hemolytic uremic syndrome (HUS) is a renal disease due to complement dysregulation. Many of the known causes of atypical HUS originate from genetic mutations of complement regulatory proteins, such as complement factor H (CFH) and thrombomodulin. ...

Abstract	Atypical hemolytic uremic syndrome (HUS) is a renal disease due to complement dysregulation. Many of the known causes of atypical HUS originate from genetic mutations of complement regulatory proteins, such as complement factor H (CFH) and thrombomodulin. However, atypical HUS has only a genetic penetrance of 40-50% of the cases and usually appears in adulthood. We introduce a novel factor that may be involved in the onset and development of atypical HUS, i.e. the endothelial surface glycocalyx. The glycocalyx is a highly interactive matrix covering the luminal side of vascular endothelial cells and consists of glycosaminoglycans, proteoglycans and glycoproteins, which has an important role in maintaining homeostasis of the vasculature. The surface-bound glycocalyx glycosaminoglycan constituent heparan sulfate is crucial for CFH binding and function, both in recognition of host tissue and prevention of spontaneous complement activation via the alternative pathway. Most of the clinically relevant genetic mutations in CFH result in incorrect binding to heparan sulfate. In addition, a role between proper function of thrombomodulin and the endothelial glycocalyx has also been observed. We suggest that not only changes in binding properties of the complement regulatory proteins play a role but also changes in the endothelial glycocalyx are involved in increased risk of clinical manifestation of atypical HUS. Finally, vascular glycocalyx heterogeneity in turn could dictate the specific vulnerability of the glomerular vascular bed in atypical HUS and may provide new therapeutic targets to intervene with endothelial cell activation and local complement pathway regulation.
MeSH term(s)	Atypical Hemolytic Uremic Syndrome ; Complement Activation/genetics ; Complement Activation/immunology ; Complement Factor H/genetics ; Complement Factor H/immunology ; Endothelium, Vascular/immunology ; Genetic Predisposition to Disease ; Glycocalyx/immunology ; Hemolytic-Uremic Syndrome/genetics ; Hemolytic-Uremic Syndrome/immunology ; Heparan Sulfate Proteoglycans/physiology ; Humans ; Thrombomodulin/genetics
Chemical Substances	Heparan Sulfate Proteoglycans ; Thrombomodulin ; Complement Factor H (80295-65-4)
Language	English
Publishing date	2013-09
Publishing country	Netherlands
Document type	Journal Article ; Review
ZDB-ID	1038679-8
ISSN	1879-0828 ; 0953-6205
ISSN (online)	1879-0828
ISSN	0953-6205
DOI	10.1016/j.ejim.2012.12.016
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Article ; Online: A microscopic view on the renal endothelial glycocalyx.

Dane, Martijn J C / van den Berg, Bernard M / Lee, Dae Hyun / Boels, Margien G S / Tiemeier, Gesa L / Avramut, M Cristina / van Zonneveld, Anton Jan / van der Vlag, Johan / Vink, Hans / Rabelink, Ton J

American journal of physiology. Renal physiology

2015 Volume 308, Issue 9, Page(s) F956–66

Abstract: Endothelial cells perform key homeostatic functions such as regulating blood flow, permeability, and aiding immune surveillance for pathogens. While endothelial activation serves normal physiological adaptation, maladaptation of these endothelial ... ...

Abstract	Endothelial cells perform key homeostatic functions such as regulating blood flow, permeability, and aiding immune surveillance for pathogens. While endothelial activation serves normal physiological adaptation, maladaptation of these endothelial functions has been identified as an important effector mechanism in the progression of renal disease as well as the associated development of cardiovascular disease. The primary interface between blood and the endothelium is the glycocalyx. This carbohydrate-rich gel-like structure with its associated proteins mediates most of the regulatory functions of the endothelium. Because the endothelial glycocalyx is a highly dynamic and fragile structure ex vivo, and traditional tissue processing for staining and perfusion-fixation usually results in a partial or complete loss of the glycocalyx, studying its dimensions and function has proven to be challenging. In this review, we will outline the core functions of the glycocalyx and focus on different techniques to study structure-function relationships in kidney and vasculature.
MeSH term(s)	Animals ; Endothelial Cells/metabolism ; Endothelial Cells/ultrastructure ; Glycocalyx/metabolism ; Glycocalyx/ultrastructure ; Humans ; Kidney/blood supply ; Kidney Diseases/metabolism ; Kidney Diseases/pathology ; Kidney Diseases/physiopathology ; Microscopy/methods ; Specimen Handling ; Staining and Labeling
Language	English
Publishing date	2015-05-01
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	603837-2
ISSN	1522-1466 ; 0363-6127
ISSN (online)	1522-1466
ISSN	0363-6127
DOI	10.1152/ajprenal.00532.2014
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Article ; Online: The RNA-binding protein quaking maintains endothelial barrier function and affects VE-cadherin and β-catenin protein expression.

de Bruin, Ruben G / van der Veer, Eric P / Prins, Jurriën / Lee, Dae Hyun / Dane, Martijn J C / Zhang, Huayu / Roeten, Marko K / Bijkerk, Roel / de Boer, Hetty C / Rabelink, Ton J / van Zonneveld, Anton Jan / van Gils, Janine M

Scientific reports

2016 Volume 6, Page(s) 21643

Abstract: Proper regulation of endothelial cell-cell contacts is essential for physiological functioning of the endothelium. Interendothelial junctions are actively involved in the control of vascular leakage, leukocyte diapedesis, and the initiation and ... ...

Abstract	Proper regulation of endothelial cell-cell contacts is essential for physiological functioning of the endothelium. Interendothelial junctions are actively involved in the control of vascular leakage, leukocyte diapedesis, and the initiation and progression of angiogenesis. We found that the RNA-binding protein quaking is highly expressed by endothelial cells, and that its expression was augmented by prolonged culture under laminar flow and the transcription factor KLF2 binding to the promoter. Moreover, we demonstrated that quaking directly binds to the mRNA of VE-cadherin and β-catenin and can induce mRNA translation mediated by the 3'UTR of these genes. Reduced quaking levels attenuated VE-cadherin and β-catenin expression and endothelial barrier function in vitro and resulted in increased bradykinin-induced vascular leakage in vivo. Taken together, we report that quaking is essential in maintaining endothelial barrier function. Our results provide novel insight into the importance of post-transcriptional regulation in controlling vascular integrity.
MeSH term(s)	Animals ; Antigens, CD/genetics ; Antigens, CD/metabolism ; Cadherins/genetics ; Cadherins/metabolism ; Capillary Permeability ; Female ; Gene Expression ; HEK293 Cells ; Human Umbilical Vein Endothelial Cells/physiology ; Humans ; Kruppel-Like Transcription Factors/physiology ; Mice, Inbred C57BL ; Mice, Transgenic ; Protein Binding ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; RNA-Binding Proteins/physiology ; Transcriptional Activation ; beta Catenin/genetics ; beta Catenin/metabolism
Chemical Substances	Antigens, CD ; CTNNB1 protein, human ; Cadherins ; KLF2 protein, human ; Kruppel-Like Transcription Factors ; QKI protein, human ; RNA, Messenger ; RNA-Binding Proteins ; beta Catenin ; cadherin 5
Language	English
Publishing date	2016-02-24
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/srep21643
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Article ; Online: Atrasentan Reduces Albuminuria by Restoring the Glomerular Endothelial Glycocalyx Barrier in Diabetic Nephropathy.

Boels, Margien G S / Avramut, M Cristina / Koudijs, Angela / Dane, Martijn J C / Lee, Dae Hyun / van der Vlag, Johan / Koster, Abraham J / van Zonneveld, Anton Jan / van Faassen, Ernst / Gröne, Hermann-Josef / van den Berg, Bernard M / Rabelink, Ton J

Diabetes

2016 Volume 65, Issue 8, Page(s) 2429–2439

Abstract: Atrasentan, a selective endothelin A receptor antagonist, has been shown to reduce albuminuria in type 2 diabetes. We previously showed that the structural integrity of a glomerular endothelial glycocalyx is required to prevent albuminuria. Therefore we ... ...

Abstract	Atrasentan, a selective endothelin A receptor antagonist, has been shown to reduce albuminuria in type 2 diabetes. We previously showed that the structural integrity of a glomerular endothelial glycocalyx is required to prevent albuminuria. Therefore we tested the potential of atrasentan to stabilize the endothelial glycocalyx in diabetic apolipoprotein E (apoE)-deficient mice in relation to its antialbuminuric effects. Treatment with atrasentan (7.5 mg/kg/day) for 4 weeks reduced urinary albumin-to-creatinine ratios by 26.0 ± 6.5% (P < 0.01) in apoE knockout (KO) mice with streptozotocin-induced diabetes consuming an atherogenic diet, without changes in gross glomerular morphology, systemic blood pressure, and blood glucose concentration. Endothelial cationic ferritin surface coverage, investigated using large-scale digital transmission electron microscopy, revealed that atrasentan treatment increases glycocalyx coverage in diabetic apoE KO mice from 40.7 ± 3.2% to 81.0 ± 12.5% (P < 0.05). This restoration is accompanied by increased renal nitric oxide concentrations, reduced expression of glomerular heparanase, and a marked shift in the balance of M1 and M2 glomerular macrophages. In vitro experiments with endothelial cells exposed to laminar flow and cocultured with pericytes confirmed that atrasentan reduced endothelial heparanase expression and increased glycocalyx thickness in the presence of a diabetic milieu. Together these data point toward a role for the restoration of endothelial function and tissue homeostasis through the antialbuminuric effects of atrasentan, and they provide a mechanistic explanation for the clinical observations of reduced albuminuria with atrasentan in diabetic nephropathy.
MeSH term(s)	Albuminuria/drug therapy ; Albuminuria/metabolism ; Animals ; Diabetes Mellitus, Experimental/drug therapy ; Diabetes Mellitus, Experimental/metabolism ; Diabetic Nephropathies/drug therapy ; Diabetic Nephropathies/metabolism ; Endothelin Receptor Antagonists/therapeutic use ; Glomerular Filtration Rate/drug effects ; Kidney Glomerulus/drug effects ; Male ; Mice ; Mice, Knockout ; Pyrrolidines/therapeutic use
Chemical Substances	Endothelin Receptor Antagonists ; Pyrrolidines ; atrasentan (V6D7VK2215)
Language	English
Publishing date	2016-08
Publishing country	United States
Document type	Journal Article
ZDB-ID	80085-5
ISSN	1939-327X ; 0012-1797
ISSN (online)	1939-327X
ISSN	0012-1797
DOI	10.2337/db15-1413
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Article ; Online: Narrowband supercontinuum control using phase shaping.

Austin, Dane R / Bolger, Jeremy A / de Sterke, C Martijn / Eggleton, Benjamin J / Brown, Thomas G

Optics express

2009 Volume 14, Issue 26, Page(s) 13142–13150

Abstract: We study theoretically, numerically and experimentally the effect of self-phase modulation of ultrashort pulses with spectrally narrow phase features. We show that spectral enhancement and depletion is caused by changing the relative phase between the ... ...

Abstract	We study theoretically, numerically and experimentally the effect of self-phase modulation of ultrashort pulses with spectrally narrow phase features. We show that spectral enhancement and depletion is caused by changing the relative phase between the initial field and the nonlinearly generated components. Our theoretical results explain observations of supercontinuum enhancement by fiber Bragg gratings, and predict similar enhancements for spectrally shaped pulses in uniform fiber. As proof of principle, we demonstrate this effect in the laboratory using a femtosecond pulse shaper.
Language	English
Publishing date	2009-08-08
Publishing country	United States
Document type	Journal Article
ZDB-ID	1491859-6
ISSN	1094-4087 ; 1094-4087
ISSN (online)	1094-4087
ISSN	1094-4087
DOI	10.1364/oe.14.013142
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Article ; Online: Dispersive wave blue-shift in supercontinuum generation.

Austin, Dane R / de Sterke, C Martijn / Eggleton, Benjamin J / Brown, Thomas G

Optics express

2009 Volume 14, Issue 25, Page(s) 11997–12007

Abstract: We numerically study dispersive wave emission during femtosecond-pumped supercontinuum generation in photonic crystal fibres. We show that dispersive waves are primarily generated over a short region of high temporal compression. Despite the apparent ... ...

Abstract	We numerically study dispersive wave emission during femtosecond-pumped supercontinuum generation in photonic crystal fibres. We show that dispersive waves are primarily generated over a short region of high temporal compression. Despite the apparent complexity of the pump pulse in this region, we show that the dynamics of dispersive wave generation are dominated by a single fundamental soliton. However, any straightforward application of the theory that is thought to describe the blue emission, considerably underestimates the frequency shift. We show that in fact the red-shift of the soliton, caused by spectral recoil from the growing dispersive wave, causes an additional blue-shift of the resonant frequency which is in good agreement with full simulations.
Language	English
Publishing date	2009-08-08
Publishing country	United States
Document type	Journal Article
ZDB-ID	1491859-6
ISSN	1094-4087 ; 1094-4087
ISSN (online)	1094-4087
ISSN	1094-4087
DOI	10.1364/oe.14.011997
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Article ; Online: Deeper penetration of erythrocytes into the endothelial glycocalyx is associated with impaired microvascular perfusion.

Lee, Dae Hyun / Dane, Martijn J C / van den Berg, Bernard M / Boels, Margien G S / van Teeffelen, Jurgen W / de Mutsert, Renée / den Heijer, Martin / Rosendaal, Frits R / van der Vlag, Johan / van Zonneveld, Anton Jan / Vink, Hans / Rabelink, Ton J

PloS one

2014 Volume 9, Issue 5, Page(s) e96477

Abstract: Changes in endothelial glycocalyx are one of the earliest changes in development of cardiovascular disease. The endothelial glycocalyx is both an important biological modifier of interactions between flowing blood and the vessel wall, and a determinant ... ...

Abstract	Changes in endothelial glycocalyx are one of the earliest changes in development of cardiovascular disease. The endothelial glycocalyx is both an important biological modifier of interactions between flowing blood and the vessel wall, and a determinant of organ perfusion. We hypothesize that deeper penetration of erythrocytes into the glycocalyx is associated with reduced microvascular perfusion. The population-based prospective cohort study (the Netherlands Epidemiology of Obesity [NEO] study) includes 6,673 middle-aged individuals (oversampling of overweight and obese individuals). Within this cohort, we have imaged the sublingual microvasculature of 915 participants using sidestream darkfield (SDF) imaging together with a recently developed automated acquisition and analysis approach. Presence of RBC (as a marker of microvascular perfusion) and perfused boundary region (PBR), a marker for endothelial glycocalyx barrier properties for RBC accessibility, were assessed in vessels between 5 and 25 µm RBC column width. A wide range of variability in PBR measurements, with a mean PBR of 2.14 µm (range: 1.43-2.86 µm), was observed. Linear regression analysis showed a marked association between PBR and microvascular perfusion, reflected by RBC filling percentage (regression coefficient β: -0.034; 95% confidence interval: -0.037 to -0.031). We conclude that microvascular beds with a thick ("healthy") glycocalyx (low PBR), reflects efficient perfusion of the microvascular bed. In contrast, a thin ("risk") glycocalyx (high PBR) is associated with a less efficient and defective microvascular perfusion.
MeSH term(s)	Body Mass Index ; Cardiovascular Diseases/diagnosis ; Cardiovascular Diseases/epidemiology ; Cross-Sectional Studies ; Diagnostic Imaging/methods ; Endothelium, Vascular/metabolism ; Erythrocytes/metabolism ; Female ; Glycocalyx/metabolism ; Humans ; Linear Models ; Male ; Microvessels/metabolism ; Middle Aged ; Netherlands/epidemiology ; Obesity/diagnosis ; Obesity/epidemiology ; Perfusion ; Population Surveillance/methods ; Prospective Studies ; Surveys and Questionnaires
Language	English
Publishing date	2014-05-09
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0096477
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Article ; Online: Association of kidney function with changes in the endothelial surface layer.

Dane, Martijn J C / Khairoun, Meriem / Lee, Dae Hyun / van den Berg, Bernard M / Eskens, Bart J M / Boels, Margien G S / van Teeffelen, Jurgen W G E / Rops, Angelique L W M M / van der Vlag, Johan / van Zonneveld, Anton Jan / Reinders, Marlies E J / Vink, Hans / Rabelink, Ton J

Clinical journal of the American Society of Nephrology : CJASN

2014 Volume 9, Issue 4, Page(s) 698–704

Abstract: Background and objectives: ESRD is accompanied by endothelial dysfunction. Because the endothelial glycocalyx (endothelial surface layer) governs interactions between flowing blood and the vessel wall, perturbation could influence disease progression. ... ...

Abstract	Background and objectives: ESRD is accompanied by endothelial dysfunction. Because the endothelial glycocalyx (endothelial surface layer) governs interactions between flowing blood and the vessel wall, perturbation could influence disease progression. This study used a novel noninvasive sidestream-darkfield imaging method, which measures the accessibility of red blood cells to the endothelial surface layer in the microcirculation (perfused boundary region), to investigate whether renal function is associated with endothelial surface layer dimensions. Design, setting, participants, & measurements: Perfused boundary region was measured in control participants (n=10), patients with ESRD (n=23), participants with normal kidney function after successful living donor kidney transplantation (n=12), and patients who developed interstitial fibrosis/tubular atrophy after kidney transplantation (n=10). In addition, the endothelial activation marker angiopoietin-2 and shed endothelial surface layer components syndecan-1 and soluble thrombomodulin were measured using ELISA. Results: Compared with healthy controls (1.82 ± 0.16 µm), ESRD patients had a larger perfused boundary region (+0.23; 95% confidence interval, 0.46 to <0.01; P<0.05), which signifies loss of endothelial surface layer dimensions. This large perfused boundary region was accompanied by higher circulating levels of syndecan-1 (+57.71; 95% confidence interval, 17.38 to 98.04; P<0.01) and soluble thrombomodulin (+12.88; 95% confidence interval, 0.29 to 25.46; P<0.001). After successful transplantation, the perfused boundary region was indistinguishable from healthy controls (without elevated levels of soluble thrombomodulin or syndecan-1). In contrast, however, patients who developed interstitial fibrosis and tubular atrophy showed a large perfused boundary region (+0.36; 95% confidence interval, 0.09 to 0.63; P<0.01) and higher levels of endothelial activation markers. In addition, a significant correlation between perfused boundary region, angiopoietin-2, and eGFR was observed (perfused boundary region versus GFR: Spearman's ρ=0.31; P<0.05; perfused boundary region versus angiopoietin-2: Spearman's ρ=-0.33; P<0.05). Conclusion: Reduced renal function is strongly associated with low endothelial surface layer dimensions. After successful kidney transplantation, the endothelial surface layer is indistinguishable from control.
MeSH term(s)	Adult ; Aged ; Angiopoietin-2/blood ; Animals ; Atrophy ; Biomarkers/blood ; Case-Control Studies ; Cross-Sectional Studies ; Endothelial Cells/metabolism ; Endothelial Cells/pathology ; Fibrosis ; Glycocalyx/pathology ; Humans ; Kidney/pathology ; Kidney/physiopathology ; Kidney/surgery ; Kidney Failure, Chronic/blood ; Kidney Failure, Chronic/pathology ; Kidney Failure, Chronic/physiopathology ; Kidney Failure, Chronic/surgery ; Kidney Transplantation/adverse effects ; Male ; Mice ; Microcirculation ; Microvessels/metabolism ; Microvessels/pathology ; Microvessels/physiopathology ; Middle Aged ; Predictive Value of Tests ; Regional Blood Flow ; Reproducibility of Results ; Syndecan-1/blood ; Thrombomodulin/blood ; Tongue/blood supply ; Treatment Outcome
Chemical Substances	ANGPT2 protein, human ; Angiopoietin-2 ; Biomarkers ; SDC1 protein, human ; Syndecan-1 ; THBD protein, human ; Thrombomodulin
Language	English
Publishing date	2014-01-23
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2226665-3
ISSN	1555-905X ; 1555-9041
ISSN (online)	1555-905X
ISSN	1555-9041
DOI	10.2215/CJN.08160813
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Article ; Online: Glomerular endothelial surface layer acts as a barrier against albumin filtration.

Dane, Martijn J C / van den Berg, Bernard M / Avramut, M Cristina / Faas, Frank G A / van der Vlag, Johan / Rops, Angelique L W M M / Ravelli, Raimond B G / Koster, Bram J / van Zonneveld, Anton Jan / Vink, Hans / Rabelink, Ton J

The American journal of pathology

2013 Volume 182, Issue 5, Page(s) 1532–1540

Abstract: Glomerular endothelium is highly fenestrated, and its contribution to glomerular barrier function is the subject of debate. In recent years, a polysaccharide-rich endothelial surface layer (ESL) has been postulated to act as a filtration barrier for ... ...

Abstract	Glomerular endothelium is highly fenestrated, and its contribution to glomerular barrier function is the subject of debate. In recent years, a polysaccharide-rich endothelial surface layer (ESL) has been postulated to act as a filtration barrier for large molecules, such as albumin. To test this hypothesis, we disturbed the ESL in C57Bl/6 mice using long-term hyaluronidase infusion for 4 weeks and monitored albumin passage using immunolabeling and correlative light-electron microscopy that allows for complete and integral assessment of glomerular albumin passage. ESL ultrastructure was visualized by transmission electron microscopy using cupromeronic blue and by localization of ESL binding lectins using confocal microscopy. We demonstrate that glomerular fenestrae are filled with dense negatively charged polysaccharide structures that are largely removed in the presence of circulating hyaluronidase, leaving the polysaccharide surfaces of other glomerular cells intact. Both retention of native ferritin [corrected] in the glomerular basement membrane and systemic blood pressure were unaltered. Enzyme treatment, however, induced albumin passage across the endothelium in 90% of glomeruli, whereas this could not be observed in controls. Yet, there was no net albuminuria due to binding and uptake of filtered albumin by the podocytes and parietal epithelium. ESL structure and function completely recovered within 4 weeks on cessation of hyaluronidase infusion. Thus, the polyanionic ESL component, hyaluronan, is a key component of the glomerular endothelial protein permeability barrier.
MeSH term(s)	Albumins/metabolism ; Animals ; Cattle ; Endothelium/drug effects ; Endothelium/physiology ; Endothelium/ultrastructure ; Fluorescence ; Glomerular Basement Membrane/drug effects ; Glomerular Basement Membrane/physiology ; Glomerular Basement Membrane/ultrastructure ; Glomerular Filtration Rate/drug effects ; Glomerular Filtration Rate/physiology ; Horses ; Hyaluronoglucosaminidase/pharmacology ; Kidney Glomerulus/cytology ; Kidney Glomerulus/drug effects ; Kidney Glomerulus/physiology ; Kidney Glomerulus/ultrastructure ; Lectins/metabolism ; Mice ; Mice, Inbred C57BL ; Permeability/drug effects ; Podocytes/cytology ; Podocytes/drug effects ; Podocytes/ultrastructure
Chemical Substances	Albumins ; Lectins ; Hyaluronoglucosaminidase (EC 3.2.1.35)
Language	English
Publishing date	2013-05
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2943-9
ISSN	1525-2191 ; 0002-9440
ISSN (online)	1525-2191
ISSN	0002-9440
DOI	10.1016/j.ajpath.2013.01.049
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