Lectin histochemical examination of rabbit bladder glycoproteins and characterization of a mucin isolated from the bladder mucosa.
The glycocalyx of the mucosal surface of urinary bladder acts as an effective barrier against invasion by pathogenic microorganisms and injury from toxic substances in the urine. Defects in these bladder mucosal components could thus be important factors in the development of diseases such as interstitial cystitis and lower urinary tract infections. However, information on the nature of glycoconjugates of mammalian bladder mucosa is very limited. In this study, the glycoconjugates of rabbit bladder were examined histochemically using biotinylated lectins with specificities for a variety of carbohydrate moieties. Three [Artocarpus integrifolia (Jacalin), Datura stramonium (DSL), and Maackia amurensis II (MAL-II)] of the lectins bound predominantly to the luminal cell layer, with decreased binding to the basal layers of the epithelium. In contrast, Ricinus communis I and Sambucus nigra lectins did not bind to the cells in the epithelium but strongly interacted with the subepithelial layers, especially the lamina propria. The intensity of the staining by Jacalin and MAL-II was significantly reduced by prior treatment of the bladder sections with O-sialoglycoprotein endopeptidase, indicating that the ligands of these lectins are primarily mucin glycoproteins. In parallel biochemical studies, a high-molecular-weight glycoprotein with characteristics typical of epithelial mucins was purified from the mucosa of rabbit bladder explant cultures metabolically labeled with [(3)H]glucosamine. Quantitative analysis of the sialic acid, uronic acid, and hexosamine contents of delipidated rabbit bladder mucosa revealed a larger proportion of sialoglycoproteins compared with glycosaminoglycans. Taken together, the results of histochemical and biochemical analyses indicate that glycoproteins rather than glycosaminoglycans are the major components of the bladder epithelium, and that the former include a mucin.
Buckley
M
M
Department of Biochemistry, Penn State University College of Medicine, Hershey, Pennsylvania, 17033, USA.
Xin
P
P
Washington
S
S
Herb
N
N
Erickson
D
D
Bhavanandan
V P
VP
D000818:Animals; D019921:Biotinylation; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D006025:Glycosaminoglycans / Q000032:analysis / Q000737:chemistry; D006595:Hexosamines / Q000032:analysis; D037102:Lectins / Q000378:metabolism*; D008666:Metalloendopeptidases / Q000378:metabolism; D008970:Molecular Weight; D009077:Mucins / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D009092:Mucous Membrane / Q000737:chemistry / Q000378:metabolism; D019158:N-Acetylneuraminic Acid / Q000032:analysis; D009924:Organ Culture Techniques; D011817:Rabbits; D012795:Sialoglycoproteins / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D001743:Urinary Bladder / Q000737:chemistry* / Q000166:cytology; D014574:Uronic Acids / Q000032:analysis
D006025:Glycosaminoglycans; D006595:Hexosamines; D037102:Lectins; D009077:Mucins; D012795:Sialoglycoproteins; D014574:Uronic Acids; D008666:Metalloendopeptidases; C070738:O-sialoglycoprotein endopeptidase; D019158:N-Acetylneuraminic Acid