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Endocrinology Vol. 142, No. 1 339-347
Copyright © 2001 by The Endocrine Society

ARTICLES

Vasoactive Intestinal Peptide (VIP)/Pituitary Adenylate Cyclase-Activating Peptide Receptor Subtypes in Mouse Calvarial Osteoblasts: Presence of VIP-2 Receptors and Differentiation-Induced Expression of VIP-1 Receptors1

Pernilla Lundberg, Inger Lundgren, Hitoshi Mukohyama, Petri P. Lehenkari, Michael A. Horton and Ulf H. Lerner

Department of Odontology (P.L., I.L., H.M., U.H.L.), Oral Cell Biology, Umeå University, Umeå, Sweden; Department of Musculoskeletal Research (P.L., I.L., U.H.L.), National Institute for Working Life, Umeå, Sweden; and Bone and Mineral Centre (P.P.L., M.A.H.), Department of Medicine, The Rayne Institute, University College London, London WC1E 6JJ, United Kingdom

Address all correspondence and requests for reprints to: Pernilla Lund-berg, Department of Odontology, Oral Cell Biology, Umeå University, Umeå, SE-901 87, Sweden. E-mail: Pernilla Lundberg{at}odont.umu.se

Three distinct complementary DNAs for vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating peptide (PACAP) receptors have been cloned and designated VIP-1 receptor (VIP-1R), VIP-2 receptor (VIP-2R), and PACAP receptor (PACAP-R). In the present study, we have characterized the binding sites on primary mouse calvarial osteoblasts for VIP and related peptides. By analyzing the cAMP response, the rank order of response observed was PACAP 38 > PACAP 27 > helodermin > VIP > helospectin > glucagon > PHI >>> secretin. The VIP-2R/PACAP-R antagonist, PACAP 6–38, inhibited both VIP- and PACAP-stimulated cAMP formation. Binding studies using an atomic force microscopy (AFM) technique showed high affinity binding for VIP and PACAP 38, but not for secretin. Radioligand binding studies using 125I-VIP and 125I-PACAP 38 demonstrated a more specific and higher affinity binding for PACAP 38 than for VIP. Secretin failed to inhibit both 125I-VIP and 125I-PACAP 38 binding. RT-PCR demonstrated that undifferentiated mouse calvarial osteoblasts express messenger RNA for VIP-2R, but not for VIP-1R or PACAP-R. When the osteoblasts were cultured for 20 days to induce bone noduli formation, VIP-1R, in addition to VIP-2R, were expressed when the nodules started to mineralize at 12 days. Taken together, these data demonstrate that mouse calvarial osteoblasts express functional VIP-2R with higher affinity binding for PACAP than for VIP and that the VIP-1R expression is induced during osteoblastic differentiation.




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