Pflügers Archiv - European Journal of Physiology

, Volume 465, Issue 2, pp 271–281

E2f1-deficient NOD/SCID mice have dry mouth due to a change of acinar/duct structure and the down-regulation of AQP5 in the salivary gland

Authors

    • Department of Regulatory PhysiologyDokkyo Medical University School of Medicine
  • Takanori Narita
    • Laboratory of Veterinary Biochemistry, College of Bioresource SciencesNihon University
  • Miwako Matsuki-Fukushima
    • Department of PhysiologyNihon University School of Dentistry at Matsudo
  • Ken Okabayashi
    • Laboratory of Veterinary Biochemistry, College of Bioresource SciencesNihon University
  • Tatsuro Ito
    • Department of Pediatric DentistryNihon University School of Dentistry at Matsudo
    • Department of BacteriologyNational Institute of Infectious Diseases
  • Hidenobu Senpuku
    • Department of BacteriologyNational Institute of Infectious Diseases
  • Hiroshi Sugiya
    • Laboratory of Veterinary Biochemistry, College of Bioresource SciencesNihon University
    • Oral Health Science Center Project, hrc8Tokyo Dental College
Molecular and Cellular Mechanisms of Disease

DOI: 10.1007/s00424-012-1183-y

Cite this article as:
Satoh, K., Narita, T., Matsuki-Fukushima, M. et al. Pflugers Arch - Eur J Physiol (2013) 465: 271. doi:10.1007/s00424-012-1183-y

Abstract

Non-obese diabetic (NOD) mice have been used as a model for dry mouth. NOD mice lacking the gene encoding E2f1, a transcription factor, develop hyposalivation more rapidly progressively than control NOD mice. However, the model mice are associated with an underlying disease such as diabetes. We have now established E2f1-deficient NOD/severe combined immunodeficiency disease (NOD/SCID.E2f1−/−) mice to avoid the development of diabetes (Matsui-Inohara et al., Exp Biol Med (Maywood) 234(12):1525–1536, 2009). In this study, we investigated the pathophysiological features of dry mouth using NOD/SCID.E2f1−/− mice. In NOD/SCID.E2f1−/− mice, the volume of secreted saliva stimulated with pilocarpine is about one third that of control NOD/SCID mice. In behavioral analysis, NOD/SCID.E2f1−/− mice drank plenty of water when they ate dry food, and the frequency and time of water intake were almost double compared with control NOD/SCID mice. Histological analysis of submandibular glands with hematoxylin–eosin stain revealed that NOD/SCID.E2f1−/− mice have more ducts than NOD/SCID mice. In western blot analysis, the expression of aquaporin 5 (AQP5), a marker of acinar cells, in parotid and in submandibular glands of NOD/SCID.E2f1−/− mice was lower than in NOD/SCID mice. Immunohistochemical analysis of parotid and submandibular acini revealed that the localization of AQP5 in NOD/SCID.E2f1−/− mice differs from that in NOD/SCID mice; AQP5 was leaky and diffusively localized from the apical membrane to the cytosol in NOD/SCID.E2f1−/− mice. The ubiquitination of AQP5 was detected in submandibular glands of NOD/SCID.E2f1−/− mice. These findings suggest that the change of acinar/duct structure and the down-regulation of AQP5 in the salivary gland cause the pathogenesis of hyposalivation in NOD/SCID.E2f1−/− mice.

Keywords

Salivary glands AQP5 protein Down-regulation Dry mouth Ubiquitin Disease model

Abbreviations

AQP5

Aquaporin 5

NOD

Non-obese diabetic

SCID

Severe combined immunodeficiency disease

SS

Sjögren’s syndrome

BSA

Bovine serum albumin

RT-PCR

Reverse transcription/polymerase chain reaction

GAPDH

Glyceraldehyde-3-phosphate dehydrogenase

B2M

β2 microglobulin

HPRT

Hypoxanthine phosphoribosyl transferase

SDS

Sodium dodecyl sulfate

PAGE

Polyacrylamide gel electrophoresis

IP

Immunoprecipitation

WB

Western blot analysis

Ub-AQP5

Ubiquitinated-AQP5

Supplementary material

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Copyright information

© Springer-Verlag Berlin Heidelberg 2012