Evidence of NK1 and NK2 tachykinin receptors and their involvement in histamine release in a murine mast cell line

• Published in: Neuropeptides (Edinburgh) Vol. 21; no. 2; pp. 65 - 72
• Main Authors: Krumins, S.A., Broomfield, C.A.
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier Ltd 01.02.1992; Elsevier
• Subjects: Amino Acid Sequence; Animals; Binding, Competitive; Biological and medical sciences; Cell Line; Cell Membrane - metabolism; Cell receptors; Cell structures and functions; Fundamental and applied biological sciences. Psychology; Histamine Release - drug effects; Histamine Release - physiology; Mast Cells - metabolism; Mice; Molecular and cellular biology; Molecular Sequence Data; Neurokinin A - pharmacology; Neuropeptide receptors; p-Methoxy-N-methylphenethylamine - pharmacology; Peptide Fragments - pharmacology; Peptides - pharmacology; Receptors, Neurotransmitter - physiology; Receptors, Tachykinin; Substance P - metabolism; Substance P - pharmacology; Substance P; Rodentia; Binding site; Neuropeptide; Histamine; Vertebrata; Mammalia; Mouse; Neuromediator; Established cell line; Peptidergic receptor; Mast cell; Tachykinin; Release
• ISSN: 0143-4179; 1532-2785
• DOI: 10.1016/0143-4179(92)90516-Y

Binding of [ 3H]substance P (SP) and histamine release were examined using a cloned mouse mast cell line. SP binding was saturable and specific. In the presence of 30 mM Na 2SO 4 50 mM Tris buffer, SP interacted with two types of binding sites with K d values of 0.3 and 40 nM. High-affinity SP binding was blocked by the inclusion of 0.5 uM of the NK1 receptor selective ligand septide in the binding mixture. Neurokinin A (NKA) evoked concentration-dependent histamine release. At concentrations in the nanomolar range, the NK1 preferring agonists SP, SP methylester and physalaemin evoked ≤5% net release of histamine, which was substantially less than the maximum effect of NKA (+37%) in the micromolar range. Pretreatment of the cells with the NK2 antagonist peptide A reduced NKA-induced histamine release. [D-Arg 1,D-Phe 5,D-Trp 7,9,Leu 11]-substance P, a putative SP antagonist, also elicited histamine release in the micromolar range, apparently acting as an agonist at the NK2 site. Compound 48 80 , N-terminal SP fragments, neurokinin B and the two selective NK2 receptor antagonists cyclo(Gln-Trp-Phe-(R)-[ANC-2]Leu-Met) (peptide A) and cyclo(Gln-Trp-Phe-Gly-Leu-Met) (peptide B) were ineffective. Although the results suggest the coexistence of functional NK1 and NK2 receptors, it appears that in this mast cell line neurokinin-induced histamine release is primarily mediated by the NK2 receptor, characterized biochemically as a low affinity binding site with a K d value of 40 nM for SP.