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Expression of Adrenomedullin and its Receptors in Human Salivary Tissue

S. Kapas1, K. Pahal1, A.T. Cruchley1, E. Hagi-Pavli1, and J.P. Hinson2,*

1 Molecular Signalling Group, Clinical Sciences Research Centre, and
2 Department of Endocrinology, Barts & the London, Queen Mary’s School of Medicine & Dentistry, Suite 12, Dominion House, Bartholomew Close, London EC1A 7BE, UK;



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Figure 1. Adrenomedullin concentrations (pmol/L) in normal human saliva and serum. Each value represents means ± SEM, n = 4. *P < 0.05 and ***P < 0.001 compared with levels in serum. {dagger} p < 0.001 compared with either parotid or submandibular alone (one-way ANOVA followed by a Dunnett’s test).

 


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Figure 2. Western blotting of synthetic human adrenomedullin and saliva. Pooled fractions of submandibular, parotid, or whole saliva were subjected to immunoblotting. Only one band, of approximately 6 kDa, was observed in all samples. Experiments were carried out 3 times, with the same result.

 


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Figure 3. Immunohistochemical localization of adrenomedullin in a human minor salivary gland. Note ductal epithelial cell staining (lower panel). Similar findings were observed in the parotid and submandibular salivary glands (not illustrated). Upper panel, negative staining control involving the primary rabbit adrenomedullin antibody absorbed with adrenomedullin antigen before application to the specimen. Scale bar = 50 µm. This image was chosen on the basis that the histology is clearly identifiable and the localization of adrenomedullin can be seen clearly.

 


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Figure 4. Scatchard analysis and displacement curves of human [125I] adrenomedullin. (A,B) Data from HSY cells. (C,D) Data from HSG cells. (A,C) Scatchard analysis of specific [125I] adrenomedullin binding; displacement with adrenomedullin. (B,D) Displacement of [125I] adrenomedullin binding by increasing concentrations of adrenomedullin (filled squares), CGRP (open squares), calcitonin (open circles), amylin (filled circles), or PAMP (filled triangles). Each point represents the mean of triplicate determinations.

 





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