The bladder does not appear to have a dynamic secreted continuous mucous gel layer

James Michael Olu N'Dow, N. Jordan, C. N. Robson, D. E. Neal, J. P. Pearson

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Purpose:
We determined whether the nature of any protective barrier in the bladder is composed of a secreted mucous gel layer.

Materials and Methods:
We collected 24-hour urine samples for analysis from 8 healthy 22 to 49-year-old volunteers and 5, 19 to 59-year-old patients treated with bladder reconstruction, in addition to scrapings from 100 freshly slaughtered pig bladders. Samples were subjected to homogenization, dialysis, freeze-drying, papain digestion, gel chromatography, equilibrium density gradient centrifugation, periodic acid-Schiff assay and amino acid analysis. Normal human bladder, pig bladder, normal ileum and transposed intestinal segments were studied for the presence of a mucous layer using a new method of histological analysis.

Results:
Mucin content in normal urine is 2.7 mg/24 hours, meaning that less than 0.6% of nondialyzable material in normal urine is mucin. The mucin content of urine from reconstructed bladders amounted to 86 mg/24 hours (5.2% of nondialyzable material). We observed that glycosaminoglycans accounted for 41% of the peak total elution volume of PAS positive material in normal urine. Mucin estimation in urine can be grossly overestimated if contaminating glycoconjugates are not removed. Biochemical analysis of material scraped off the pig bladder surface demonstrated that the maximum thickness of a continuous layer that could be achieved was 13.6 µm. While we could visualize an obvious mucous layer on control ileal samples and biopsies of transposed ileal segments from patients with bladder reconstruction, we were unable to note a distinct, measurable mucous layer lining the bladder surface in humans or pigs.

Conclusions:
Mucin levels in normal human and pig urine would be enough for slow turnover of a thin barrier but the large increase in mucin in the urine of patients with transposed intestinal segments demonstrates that any layer in normal bladder is much different than that lining the transposed intestinal segment. The most likely constituents of this barrier are membrane bound rather than secreted mucins along with the proteoglycan components of the glycocalix.


Original languageEnglish
Pages (from-to)2025-2031
Number of pages6
JournalJournal of Urology
Volume173
Issue number6
DOIs
Publication statusPublished - Jun 2005

Fingerprint

Urinary Bladder
Gels
Mucins
Urine
Swine
Glycocalyx
Periodic Acid
Glycoconjugates
Papain
Freeze Drying
Density Gradient Centrifugation
Proteoglycans
Glycosaminoglycans
Ileum
Gel Chromatography
Dialysis
Digestion
Volunteers
Biopsy
Amino Acids

Keywords

  • bladder
  • urinary diversion
  • mucins
  • mucus
  • urothelium

Cite this

The bladder does not appear to have a dynamic secreted continuous mucous gel layer. / N'Dow, James Michael Olu; Jordan, N.; Robson, C. N.; Neal, D. E.; Pearson, J. P.

In: Journal of Urology, Vol. 173, No. 6, 06.2005, p. 2025-2031.

Research output: Contribution to journalArticle

N'Dow, James Michael Olu ; Jordan, N. ; Robson, C. N. ; Neal, D. E. ; Pearson, J. P. / The bladder does not appear to have a dynamic secreted continuous mucous gel layer. In: Journal of Urology. 2005 ; Vol. 173, No. 6. pp. 2025-2031.
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T1 - The bladder does not appear to have a dynamic secreted continuous mucous gel layer

AU - N'Dow, James Michael Olu

AU - Jordan, N.

AU - Robson, C. N.

AU - Neal, D. E.

AU - Pearson, J. P.

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N2 - Purpose: We determined whether the nature of any protective barrier in the bladder is composed of a secreted mucous gel layer. Materials and Methods: We collected 24-hour urine samples for analysis from 8 healthy 22 to 49-year-old volunteers and 5, 19 to 59-year-old patients treated with bladder reconstruction, in addition to scrapings from 100 freshly slaughtered pig bladders. Samples were subjected to homogenization, dialysis, freeze-drying, papain digestion, gel chromatography, equilibrium density gradient centrifugation, periodic acid-Schiff assay and amino acid analysis. Normal human bladder, pig bladder, normal ileum and transposed intestinal segments were studied for the presence of a mucous layer using a new method of histological analysis. Results: Mucin content in normal urine is 2.7 mg/24 hours, meaning that less than 0.6% of nondialyzable material in normal urine is mucin. The mucin content of urine from reconstructed bladders amounted to 86 mg/24 hours (5.2% of nondialyzable material). We observed that glycosaminoglycans accounted for 41% of the peak total elution volume of PAS positive material in normal urine. Mucin estimation in urine can be grossly overestimated if contaminating glycoconjugates are not removed. Biochemical analysis of material scraped off the pig bladder surface demonstrated that the maximum thickness of a continuous layer that could be achieved was 13.6 µm. While we could visualize an obvious mucous layer on control ileal samples and biopsies of transposed ileal segments from patients with bladder reconstruction, we were unable to note a distinct, measurable mucous layer lining the bladder surface in humans or pigs. Conclusions: Mucin levels in normal human and pig urine would be enough for slow turnover of a thin barrier but the large increase in mucin in the urine of patients with transposed intestinal segments demonstrates that any layer in normal bladder is much different than that lining the transposed intestinal segment. The most likely constituents of this barrier are membrane bound rather than secreted mucins along with the proteoglycan components of the glycocalix.

AB - Purpose: We determined whether the nature of any protective barrier in the bladder is composed of a secreted mucous gel layer. Materials and Methods: We collected 24-hour urine samples for analysis from 8 healthy 22 to 49-year-old volunteers and 5, 19 to 59-year-old patients treated with bladder reconstruction, in addition to scrapings from 100 freshly slaughtered pig bladders. Samples were subjected to homogenization, dialysis, freeze-drying, papain digestion, gel chromatography, equilibrium density gradient centrifugation, periodic acid-Schiff assay and amino acid analysis. Normal human bladder, pig bladder, normal ileum and transposed intestinal segments were studied for the presence of a mucous layer using a new method of histological analysis. Results: Mucin content in normal urine is 2.7 mg/24 hours, meaning that less than 0.6% of nondialyzable material in normal urine is mucin. The mucin content of urine from reconstructed bladders amounted to 86 mg/24 hours (5.2% of nondialyzable material). We observed that glycosaminoglycans accounted for 41% of the peak total elution volume of PAS positive material in normal urine. Mucin estimation in urine can be grossly overestimated if contaminating glycoconjugates are not removed. Biochemical analysis of material scraped off the pig bladder surface demonstrated that the maximum thickness of a continuous layer that could be achieved was 13.6 µm. While we could visualize an obvious mucous layer on control ileal samples and biopsies of transposed ileal segments from patients with bladder reconstruction, we were unable to note a distinct, measurable mucous layer lining the bladder surface in humans or pigs. Conclusions: Mucin levels in normal human and pig urine would be enough for slow turnover of a thin barrier but the large increase in mucin in the urine of patients with transposed intestinal segments demonstrates that any layer in normal bladder is much different than that lining the transposed intestinal segment. The most likely constituents of this barrier are membrane bound rather than secreted mucins along with the proteoglycan components of the glycocalix.

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KW - mucins

KW - mucus

KW - urothelium

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DO - 10.1097/01.ju.0000158454.47299.ae

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JO - Journal of Urology

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