Antipeptide Polyclonal Antibodies that Recognize a Substance P-Binding Site in Mammalian Tissues: A Biochemical and Immunocytochemical Study
Version of Record online: 23 NOV 2002
Journal of Neurochemistry
Volume 63, Issue 1, pages 333–343, July 1994
How to Cite
Bret-Dibat, J. L., Zouaoui, D., Déry, O., Zerari, F., Grassi, J., Maillet, S., Conrath, M. and Couraud, J. Y. (1994), Antipeptide Polyclonal Antibodies that Recognize a Substance P-Binding Site in Mammalian Tissues: A Biochemical and Immunocytochemical Study. Journal of Neurochemistry, 63: 333–343. doi: 10.1046/j.1471-4159.1994.63010333.x
- Issue online: 23 NOV 2002
- Version of Record online: 23 NOV 2002
- Received August 17, 1993; revised manuscript received December 1, 1993; accepted December 1, 1993.
- NK1 receptor;
- Substance P;
- Complementary peptide;
- Parotid gland;
- Spinal cord
Abstract: We used complementary peptide methodology to obtain antibodies against the receptor for the neuropeptide substance P, specifically directed at the ligand-binding domain. Rabbits were immunized with two distinct peptides derived from the sequence of the RNA complementary to the mRNA for substance P. Binding experiments revealed that antipeptide polyclonal antibodies were able to recognize, through their paratope, a specific binding site on the rat parotid cell membranes. Substance P and antibodies competed for this binding site, because preincubation of membranes in the presence of substance P significantly reduced antibody binding, and conversely, preincubation of membranes in the presence of antibodies partly inhibited the binding of radioiodinated substance P. Immunocytochemical experiments performed on the rat cervical spinal cord show that the distribution of labeling by antibodies is similar to that observed by conventional autoradiography using 125I-substance P. Here again, control experiments demonstrated that antibodies and substance P were competing for the same binding site on the spinal cord. These biochemical and immunocytochemical data indicate that antipeptide antibodies recognize a substance P membrane binding site in nervous and nonnervous mammalian tissues. This site is likely to correspond to the NK1 specific receptor for substance P.