The Detection of Glycosaminoglycans in Pancreatic Islets and Lymphoid Tissues

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2014 Oct 19

PMID 25325969

Citations 6

Authors

Marika Bogdani

Charmaine Simeonovic

Nadine Nagy

Pamela Y Johnson

Christina K Chan

Thomas N Wight

Affiliations

Soon will be listed here.

Abstract

In this chapter, we describe the detection of the glycosaminoglycans hyaluronan and heparan sulfate in pancreatic islets and lymphoid tissues. The identification of hyaluronan in tissues is achieved by utilizing a highly specific hyaluronan binding protein (HABP) probe that interacts with hyaluronan in tissue sections. The HABP probe is prepared by enzymatic digestion of the chondroitin sulfate proteoglycan aggrecan which is present in bovine nasal cartilage, and is then biotinylated in the presence of bound hyaluronan and the link protein. Hyaluronan is then removed by gel filtration chromatography. The biotinylated HABP-link protein complex is applied to tissue sections and binding of the complex to tissue hyaluronan is visualized by enzymatic precipitation of chromogenic substrates. To determine hyaluronan content in tissues, tissues are first proteolytically digested to release hyaluronan from the macromolecular complexes that this molecule forms with other extracellular matrix constituents. Digested tissue is then incubated with HABP. The hyaluronan-HABP complexes are extracted and the hyaluronan concentration in the tissue is determined using an ELISA-like assay. Heparan sulfate is identified in mouse tissues by Alcian blue histochemistry and indirect immunohistochemistry. In human tissues, heparan sulfate is best detected by indirect immunohistochemistry using a specific anti-heparan sulfate monoclonal antibody. A biotinylated secondary antibody is then applied in conjunction with streptavidin-peroxidase and its binding to the anti-heparan sulfate antibody is visualized by enzymatic precipitation of chromogenic substrates.

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References

Tan K, McGrouther D, Day A, Milner C, Bayat A . Characterization of hyaluronan and TSG-6 in skin scarring: differential distribution in keloid scars, normal scars and unscarred skin. J Eur Acad Dermatol Venereol. 2010; 25(3):317-27. PMC: 3504979. DOI: 10.1111/j.1468-3083.2010.03792.x. View

Ripellino J, Bailo M, Margolis R, Margolis R . Light and electron microscopic studies on the localization of hyaluronic acid in developing rat cerebellum. J Cell Biol. 1988; 106(3):845-55. PMC: 2115103. DOI: 10.1083/jcb.106.3.845. View

Parkkinen J, Hakkinen T, Savolainen S, Wang C, Tammi R, Agren U . Distribution of hyaluronan in articular cartilage as probed by a biotinylated binding region of aggrecan. Histochem Cell Biol. 1996; 105(3):187-94. DOI: 10.1007/BF01462291. View

Kramer R, Rosen S, McDonald K . Basement-membrane components associated with the extracellular matrix of the lymph node. Cell Tissue Res. 1988; 252(2):367-75. DOI: 10.1007/BF00214379. View

Boregowda R, Appaiah H, Siddaiah M, Kumarswamy S, Sunila S, Thimmaiah K . Expression of hyaluronan in human tumor progression. J Carcinog. 2006; 5:2. PMC: 1360664. DOI: 10.1186/1477-3163-5-2. View

Lewis A, Steadman R, Manley P, Craig K, de la Motte C, Hascall V . Diabetic nephropathy, inflammation, hyaluronan and interstitial fibrosis. Histol Histopathol. 2008; 23(6):731-9. DOI: 10.14670/HH-23.731. View

Tammi R, Ripellino J, Margolis R, Tammi M . Localization of epidermal hyaluronic acid using the hyaluronate binding region of cartilage proteoglycan as a specific probe. J Invest Dermatol. 1988; 90(3):412-4. DOI: 10.1111/1523-1747.ep12456530. View

Underhill C, Zhang L . Analysis of hyaluronan using biotinylated hyaluronan-binding proteins. Methods Mol Biol. 2000; 137:441-7. DOI: 10.1385/1-59259-066-7:441. View

de la Motte C, Drazba J . Viewing hyaluronan: imaging contributes to imagining new roles for this amazing matrix polymer. J Histochem Cytochem. 2011; 59(3):252-7. PMC: 3201155. DOI: 10.1369/0022155410397760. View

10.

Anttila M, Tammi R, Tammi M, Syrjanen K, Saarikoski S, Kosma V . High levels of stromal hyaluronan predict poor disease outcome in epithelial ovarian cancer. Cancer Res. 2000; 60(1):150-5. View

11.

Underhill C, Nguyen H, Shizari M, Culty M . CD44 positive macrophages take up hyaluronan during lung development. Dev Biol. 1993; 155(2):324-36. DOI: 10.1006/dbio.1993.1032. View

12.

Ripellino J, Klinger M, Margolis R, Margolis R . The hyaluronic acid binding region as a specific probe for the localization of hyaluronic acid in tissue sections. Application to chick embryo and rat brain. J Histochem Cytochem. 1985; 33(10):1060-6. DOI: 10.1177/33.10.4045184. View

13.

Irving-Rodgers H, Ziolkowski A, Parish C, Sado Y, Ninomiya Y, Simeonovic C . Molecular composition of the peri-islet basement membrane in NOD mice: a barrier against destructive insulitis. Diabetologia. 2008; 51(9):1680-8. PMC: 2516190. DOI: 10.1007/s00125-008-1085-x. View

14.

Kaldjian E, Gretz J, Anderson A, Shi Y, Shaw S . Spatial and molecular organization of lymph node T cell cortex: a labyrinthine cavity bounded by an epithelium-like monolayer of fibroblastic reticular cells anchored to basement membrane-like extracellular matrix. Int Immunol. 2001; 13(10):1243-53. DOI: 10.1093/intimm/13.10.1243. View

15.

Hull R, Johnson P, Braun K, Day A, Wight T . Hyaluronan and hyaluronan binding proteins are normal components of mouse pancreatic islets and are differentially expressed by islet endocrine cell types. J Histochem Cytochem. 2012; 60(10):749-60. PMC: 3524560. DOI: 10.1369/0022155412457048. View

16.

Calvitti M, Baroni T, Calastrini C, Lilli C, Caramelli E, Becchetti E . Bronchial branching correlates with specific glycosidase activity, extracellular glycosaminoglycan accumulation, TGF beta(2), and IL-1 localization during chick embryo lung development. J Histochem Cytochem. 2004; 52(3):325-34. DOI: 10.1177/002215540405200303. View

17.

Aytekin M, Comhair S, de la Motte C, Bandyopadhyay S, Farver C, Hascall V . High levels of hyaluronan in idiopathic pulmonary arterial hypertension. Am J Physiol Lung Cell Mol Physiol. 2008; 295(5):L789-99. PMC: 2584887. DOI: 10.1152/ajplung.90306.2008. View

18.

Artwohl J, Brown P, Corning B, Stein S . Report of the ACLAM Task Force on Rodent Euthanasia. J Am Assoc Lab Anim Sci. 2006; 45(1):98-105. View

19.

Toole B, Yu Q, Underhill C . Hyaluronan and hyaluronan-binding proteins. Probes for specific detection. Methods Mol Biol. 2001; 171:479-85. DOI: 10.1385/1-59259-209-0:479. View

20.

Selbi W, de la Motte C, Hascall V, Day A, Bowen T, Phillips A . Characterization of hyaluronan cable structure and function in renal proximal tubular epithelial cells. Kidney Int. 2006; 70(7):1287-95. DOI: 10.1038/sj.ki.5001760. View