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Measurement of Fluid Secretion from Intact Airway Submucosal Glands

  • Jeffrey J. WineEmail author
  • Nam Soo Joo
  • Jae Young Choi
  • Hyung-Ju Cho
  • Mauri E. Krouse
  • Jin V. Wu
  • Monal Khansaheb
  • Toshiya Irokawa
  • Juan Ianowski
  • John W. Hanrahan
  • Alan W. Cuthbert
  • Kim V. Tran
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 742)

Abstract

Human airways are kept sterile by a mucosal innate defense system that includes mucus secretion. Mucus is secreted in healthy upper airways primarily by submucosal glands and consists of defense molecules mixed with mucins, electrolytes, and water and is also a major component of sputum. Mucus traps pathogens and mechanically removes them via mucociliary clearance while inhibiting their growth via molecular (e.g., lysozyme) and cellular (e.g., neutrophils, macrophages) defenses. Fluid secretion rates of single glands in response to various mediators can be measured by trapping the primary gland mucus secretions in an oil layer, where they form spherical bubbles that can be optically measured at any desired interval to provide detailed temporal analysis of secretion rates. The composition and properties of the mucus (e.g., solids, viscosity, pH) can also be determined. These methods have now been applied to mice, ferrets, cats, pigs, sheep, and humans, with a main goal of comparing gland secretion in control and CFTR-deficient humans and animals.

Key words

Submucosal gland exocrine secretion CFTR mucosal innate defense ion channel ion transporter calcium-activated chloride channel (CaCC) cAMP Ca2+ myoepithelial cell serous cell mucous cell 

Notes

Acknowledgments

We are grateful to the transplant patients and their families whose cooperation provided the tissues needed for studies of human gland secretion. For help in obtaining informed consent from patients we thank D. Weill, N.R. Henig, J. Theodore, T.E. Robinson, M. Wine, and K. Tran. For access to surgical tissues we thank B.A. Reitz, G.J. Berry, R.C. Robbins, R.I. Whyte, and the staff of the Stanford Transplant team. Jennifer Lyons provided useful discussions and comments. Technical help and data analysis were provided by Tracy Hsu, Christina Tseng, Molly Pam, Wei Chen, Sidney Chang, Kim Tran, and Jonathan Chen. The work was supported by NIH Grant DK-51817, the Cystic Fibrosis Foundation, and CFRI.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Jeffrey J. Wine
    • 1
    Email author
  • Nam Soo Joo
    • 2
  • Jae Young Choi
    • 2
    • 3
  • Hyung-Ju Cho
    • 2
  • Mauri E. Krouse
    • 2
  • Jin V. Wu
    • 2
  • Monal Khansaheb
    • 2
  • Toshiya Irokawa
    • 4
  • Juan Ianowski
    • 5
  • John W. Hanrahan
    • 6
  • Alan W. Cuthbert
    • 7
  • Kim V. Tran
    • 2
  1. 1.Cystic Fibrosis Research LaboratoryStanford UniversityStanfordUSA
  2. 2.Cystic Fibrosis Research LaboratoryStanford UniversityStanfordUSA
  3. 3.Department of OtorhinolaryngologyYonsei UniversitySeoulKorea
  4. 4.Health Administration CenterTohoku UniversitySendaiJapan
  5. 5.Department of PhysiologyUniversity of SaskatchewanSaskatoonCanada
  6. 6.Department of PhysiologyMcGill UniversityMontrealCanada
  7. 7.Department of MedicineUniversity of Cambridge, Addenbrooke’s HospitalCambridgeUK

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