C. Stratowa et al., FUNCTIONAL-CHARACTERIZATION OF THE HUMAN NEUROKININ RECEPTORS NK1, NK2, AND NK3 BASED ON A CELLULAR-ASSAY SYSTEM, Journal of receptor and signal transduction research, 15(1-4), 1995, pp. 617-630
The neurokinin receptor family is known to modulate phospholipase C ac
tivity. In order to find new compounds modulating the activity of thes
e receptors we have developed a cellular screening system that measure
s the biological activity of receptors coupled to the IP3/DAG signal t
ransduction pathway via the transcriptional activation of a reporter g
ene. For the establishment of neurokinin test cell lines the reporter
cell line A20, stably transformed with the luciferase gene under the c
ontrol of a promoter containing TPA response elements (TRE), which did
not respond to neurokinin agonists, was used. Stable test cell lines
were developed by transfecting the reporter cell line A20 with the gen
es for the human neurokinin receptors NK1, NK2 or NK3, respectively. I
n these cell lines, expression of luciferase was inducible by substanc
e P, neurokinin A and neurokinin B, respectively. The order of potency
of the three neurokinins substance P, neurokinin A and neurokinin B w
as consistent with published data and results from ligand binding stud
ies performed with the NK1 and NK2 test cell lines. The agonistic effe
ct of the neurokinins could be inhibited in a dose-dependent manner by
simultaneous addition of neurokininspecific antagonists like the non-
peptide antagonists CP-99,994 and SR 48968.