Desensitization of histamine H₁ receptor‐mediated inositol phosphate production in HeLa cells

Abstract

1 Histamine stimulated the accumulation of total [3H]-inositol phosphates (IPn) in control HeLa cells with an EC50 of 3.7 ± 0.7 JAM in the presence of 10 mM LiCl. The maximum response to histamine after 15 min incubation was 43 ± 5% over basal accumulation and occurred at a concentration of 1 mM histamine.2 The histamine-induced IPX, production in HeLa cells was confirmed as H, receptor-mediated, since the H, antagonist mepyramine (10-6 M) inhibited the histamine response (10-4 M) by 83 ± 7%, whereas the H2 antagonist, ranitidine (10-4M), and H3 antagonist, thioperamide (10-6M), were ineffective. 3 Histamine (10-M) pretreatment of HeLa cells for 30 min desensitized the subsequent histamineinduced IP,, accumulation. The desensitized cells accumulated IP. in response to histamine with an EC50 of 1.7 ± 0.7 JAM after 15 min incubation. The maximum histamine-induced IP,, accumulation at I0-4 M was 19 ± 5% over basal and was significantly lower (P <0.03) than the maximum response in control cells. 4 The desensitization of histamine-induced IP,, accumulation was time-dependent and, at a desensitizing histamine concentration of 10-4 M, the half-maximal attenuation occurred after approximately 9 min and maximum desensitization was achieved by 15-20 min. The desensitization of the IP,, accumulation was a reversible phenomenon and full recovery of the response occurred 150 min after the removal of the desensitizing histamine-containing medium. The half-time for the recovery of the histamine-induced response was estimated at 120 min.5 Bradykinin stimulated IP,, accumulation in HeLa cells, and the ECm in control cells of 1.9 ± 0.2 nM was not significantly different from the EC50 value from histamine-pretreated cells of 1.6 ± 0.9 nM. The bradykinin response at 1 ILM was 194 ± 48% over basal IP,, accumulation in control cells and this value was significantly different (P <0.04) from the 1 JAM bradykinin-induced IP,, accumulation in histaminepretreated HeLa cells of 143 ± 38% over basal. 6 NaF stimulated IP,, accumulation in control HeLa cells in a dose-related manner, with the maximum effect occurring at 15-20 mM. The ECm value for NaF-stimulated IP,, accumulation in control cells was 10.5 ± 1.1 mm and the maximum response was 136 ± 41 % over basal after 20 min incubation. In histamine desensitized HeLa cells the EC50 value for NaF was 12.3 ± 0.4 mM after 20 min stimulation, which was not significantly different from the value obtained in control cells. The maximum NaFstimulated IP,, formation in desensitized cells of 68 ± 23% over basal occurred at 15 -20 mM and was significantly lower (P<0.01) than that obtained in control cells.7 We show here that the acute histamine pretreatment of HeLa cells results in the desensitization of histamine H, receptor-mediated IP,, production. The desensitization was not restricted to the H, receptor-mediated signal transduction pathway, but also includes both the bradykinin-and NaFmediated responses, supporting a heterologous desensitization mechanism. Our results are consistent with the si...

Agonist‐induced desensitization of histamine Hi receptor‐mediated inositol phospholipid hydrolysis in human umbilical vein endothelial cells

“…Qualitatively, the regulation of HI receptor mediated inositol phosphate formation in HUVEC appears to have similar characteristics to those observed in some other cell types. For example, the regulation of histamine-induced inositol phosphate formation in HUVEC by phorbol esters, and the reversal of this effect by inhibitors of protein kinase C such as staurosporine, displays very similar characteristics to the responses seen in cultured airway smooth muscle cells, HeLa cells and DDT1MF-2 cells (Bristow & Zamani, 1993;Daykin et al, 1993;Hamilton & Sims, 1987;Dickenson & Hill, 1993). In most of these cell systems, however, the histamine-induced HI-receptor desensitization appears to be independent of protein kinase C. However the role of other protein kinases has not been investigated in these other cell types.…”

“…A number of histamine H1 receptor-mediated responses in isolated tissues and immortalised cell lines have been shown to desensitize following prolonged agonist exposure (Taylor & Richelson, 1979;Donaldson & Hill, 1985;Lurie et al, 1985;Brown et al, 1986;Leurs et.al., 1990;Bristow & Zamani, 1993;Daykin et al, 1993). The mechanisms underlying this process are poorly defined and in particular controversy exists over the potential role of protein kinase C. In order to investigate some of the mechanisms underlying desensitization of the human histamine HI receptor in a non-transformed cell line, we have studied the regulation of histamine-induced inositol phospholipid hydrolysis in human umbilical vein endothelial cell (HUVEC).…”

“…Similarly, both al-adrenoceptors (in smooth muscle cells) and cholecystokinin receptors (in pancreatic acinar cells) are phosphorylated and desensitize following short term agonist exposure (Leeb-Lundberg et al, 1987;Klueppelberg et al, 1991). However, these effects all occur rapidly following agonist exposure and less is known about the mechanisms underlying the longer term effects of agonist on the regulation of PLC-coupled receptors.A number of histamine H1 receptor-mediated responses in isolated tissues and immortalised cell lines have been shown to desensitize following prolonged agonist exposure (Taylor & Richelson, 1979;Donaldson & Hill, 1985;Lurie et al, 1985;Brown et al, 1986; Leurs et.al., 1990;Bristow & Zamani, 1993;Daykin et al, 1993). The mechanisms underlying this process are poorly defined and in particular controversy exists over the potential role of protein '."…”

Desensitization of histamine H1 receptor‐mediated inositol phosphate accumulation in guinea pig cerebral cortex slices

“…After removal of histamine the desensitized tissue had recovered to 84% of the control response by 120 min and to 90% by 150 min and was not significantly different from the histamine response measured in the control slices (at 120 min). (Bristow & Young, 1991;Bristow & Zamani, 1993) and the P2y purinoceptor-coupled phosphoinositidase C system in Turkey erythrocytes (Martin & Harden, 1989 Histamine pretreatment of rat cerebellar granule cells (Dillon-Carter & Chuang, 1989), NlE-115 mouse neuroblastoma cells (Taylor & Richelson, 1979), HeLa cells (Bristow & Zamani, 1993), BC3H-1 muscle cells (Brown et al, 1986), 1321N1 human astrocytoma cells (McDonough et al, 1988), DDT, MF-2 cells (Cowlen et al, 1990), guinea-pig ileum (Donaldson & Hill, 1986), rabbit aorta (Lurie et al, 1985), guinea-pig jejunum (Leurs et al, 1990) and also of mouse cerebral cortex slices (Quach et al, 1981) (Mitsuhashi & Payan, 1988;Murray et al, 1989;Cowlen et al, 1990) Here we show that HI receptor desensitization in guineapig cerebral cortex involves a reduction in the maximum histamine-induced response, an effect that has also been found in NIE-115 neuroblastoma cells (Taylor & Richelson, 1979), BC3H-l muscle cells (Brown et al, 1986), DDT, MF-2 smooth muscle cell line (Cowlen et al, 1990) and HeLa cells (Bristow & Zamani, 1993). We observe about a 60% desensitization of the 10-3 M histamine-stimulated [3H]-IPn production, which is of similar magnitude to the degree of desensitization of response observed in mouse brain slices (Quach et al, 1981), NIE-115 cells (Taylor & Richelson, 1979) and DDT, MF-2 smooth muscle cells (Cowlen et al, 1990).…”

“…For instance, homologous desensitization occuring at the level of the HI receptor has been shown to occur in the DDT1 MF-2 smooth muscle cell line (Cowlen et al, 1990) and mouse cerebral cortex slices (Quach et al, 1981), whereas the heterologous type of receptor desensitization involving the HI receptor and the attenuation of other receptor-mediated events after prolonged histamine pretreatment has also been reported in HeLa cells (Bristow & Zamani, 1993), BC3H-1 smooth muscle cells (Brown et al, 1986) and 1321N1 human astrocytoma cells (McDonough et al, 1988 (Bristow and Banford, unpublished observation) and the mechanism of histamineinduced desensitization must remain a target for future studies.…”