Melanocortin analogue Org2766 binds to rat Schwann cells, upregulates NGF low-affinity receptor p75, and releases neurotrophic activity.
Binding of the stable melanocortin(4-9) analogue, Org2766 [Met(O2)-Glu-His-Phe-D-Lys-Phe] to cultured rat sciatic nerve Schwann cells was demonstrated using a biotinylated derivative in semiquantitative histochemical and CELISA assays. Org2766 bound to Schwann cells, but not to fibroblasts, and was displaced maximally by unlabeled Org2766, alpha-MSH and ACTH(1-24). Displacement of Org2766 from the binding sites was considerably reduced by N- and C-truncation of the peptide. Specific binding of Org2766 was also demonstrated in the immortal rat Schwann cell line SCL4.1/F7 and was more pronounced in cells displaying a differentiated morphology. Org2766 and alpha-MSH increased cyclic AMP content of Schwann cells but neither stimulated DNA synthesis when applied alone. However, in the presence of a priming (subthreshold) concentration of the mitogen, cholera toxin, Org2766 and alpha-MSH caused a delayed increase in DNA synthesis. Org2766 did not modulate the expression of several differentiation-related Schwann cell markers. However, Org2766 increased immunoreactivity for p75 low-affinity NGF receptor on Schwann cells and evoked the release of neurotrophic factor(s) that synergized with NGF in stimulating neurite outgrowth in rat DRG neurons. The results indicate that Schwann cells are a primary target for the action of Org2766 and provide evidence for an indirect mechanism by which melanocortins might stimulate neurite sprouting in regenerating peripheral nerve axons.
Dyer
J K
JK
School of Biological Sciences, University of Bristol, UK.
Philipsen
H L
HL
Tonnaer
J A
JA
Hermkens
P H
PH
Haynes
L W
LW
D000324:Adrenocorticotropic Hormone / Q000031:analogs & derivatives* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D002772:Cholera Toxin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D004247:DNA / Q000096:biosynthesis; D004357:Drug Synergism; D008969:Molecular Sequence Data; D009414:Nerve Growth Factors / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism*; D051381:Rats; D020800:Receptor, Nerve Growth Factor; D018013:Receptors, Neuropeptide / Q000378:metabolism*; D012583:Schwann Cells / Q000378:metabolism*; D013268:Stimulation, Chemical; D015854:Up-Regulation
D009414:Nerve Growth Factors; D010446:Peptide Fragments; D020800:Receptor, Nerve Growth Factor; D018013:Receptors, Neuropeptide; C013765:Org 2766; D000324:Adrenocorticotropic Hormone; D004247:DNA; D002772:Cholera Toxin; D000242:Cyclic AMP