Identification of Protein Kinase C Phosphorylation Sites Involved in Phorbol Ester-Induced Desensitization of the Histamine H1 Receptor

The histamine H1 receptor (H1R)-mediated signaling cascade is inhibited by phorbol ester-induced protein kinase C (PKC) activation. Cloning studies of the H1Rs have shown that several potential PKC phosphorylation sites are located in the third intracellular loop of H1R. To elucidate the molecular mechanism of PKC-mediated desensitization, we identified amino acid residues that are involved in the desensitization of the H1R. Two amino acid residues (Ser396, Ser398) were determined to be PKC phosphorylation sites by in vitro phosphorylation studies using a series of synthetic peptides. Treatment with phorbol ester decreased histamine-induced accumulation of inositol phosphates in Chinese hamster ovary cells expressing the H1R with a rightward shift in the EC50 value, which implies the uncoupling of the receptor from the G protein. Site-directed mutagenesis studies showed that substitution of alanine for Ser398 but not for Ser396 markedly attenuated the effect of phorbol ester, which suggests that the Ser398 residue was primarily involved in PKC-mediated desensitization.


Gene expression of histamine H1 receptor in guinea pig primary sensory neurons: a relationship between H1 receptor mRNA-expressing neurons and peptidergic neurons

Pharmacological studies have suggested that a subgroup of primary sensory neurons is responsive to histamine via the histamine H1 receptor. We addressed this issue using in situ hybridization histochemistry with a cRNA probe for the guinea pig H1 receptor gene. About 15% of the trigeminal and lumber dorsal root ganglion (DRG) neurons, but none of nodose ganglion neurons, were intensely labeled with this probe. The H1 receptor mRNA-positive neurons were exclusively small in size, and were demonstrated to give rise to unmyelinated fibers by ultrastructural analysis of isolectin B4-labeling. However, the H1 receptor mRNA-expressing DRG neurons were not immunoreactive to substance P (SP) and calcitonin gene-related peptide (CGRP). A marked increase in the number of mRNA-positive DRG neurons were observed 1-5 days after a crush injury of the sciatic nerve (3-4-fold of the control value). These neurons turned mRNA-positive after the nerve crush were also mainly small-sized. The mRNA signals were detected in many peptidergic (SP/CGRP) neurons, in contrast to the normal state. On the other hand, in the neurons which showed intense labeling in the normal condition, the mRNA signals were down-regulated. These results suggest that primary sensory neurons include two kinds of H1 receptor-expressing sensory neurons, one expressing H1 receptor mRNAs in the normal state and the other up-regulating the mRNAs following the peripheral nerve damage.

Brain Research. Molecular Brain Research


Postnatal expression of H1receptor mRNA in the rat brain: correlation to L-histidine decarboxylase expression and local upregulation in limbic seizures

Histamine is implicated in the regulation of brain functions through three distinct receptors. Endogenous histamine in the brain is derived from mast cells and neurons, but the importance of these two pools during early postnatal development is still unknown. The expression of histamine H1-receptor in the rat brain was examined using in situ hybridization during postnatal development and in adults. For comparison, the expression of L-histidine decarboxylase (HDC) in the two pools was revealed. H1-receptor was evenly expressed throughout the brain on the first postnatal days, but resembled the adult, uneven pattern already on postnatal day 5 (P5). HDC was expressed in both mast cells and tuberomammillary neurons from birth until P5, after which the mast cell expression was no more detectable. In adult rat brain, high or moderate levels of H1-receptor expression were found in the hippocampus, zona incerta, medial amygdaloid nucleus and reticular thalamic nucleus. In most areas of the adult brain the expression of H1-receptor mRNA correlates well with binding data and histaminergic innervation. A notable exception is the hypothalamus, with high fibre density but moderate or low H1-receptor expression. Systemic kainic acid administration induced increased expression of H1-receptor mRNA in the caudate-putamen and dentate gyrus, whereas no change was seen in the hippocampal subfields CA1-CA3 or in the entorhinal cortex 6 h after kainic acid injections. This significant increase supports the concept that histaminergic transmission, through H1-receptor, is involved in the regulation of seizure activity in the brain.


In situ detection of H1-receptor mRNA and absence of apoptosis in the transient histamine system of the embryonic rat brain

In the developing brain, histamine is one of the first neurotransmitters to appear. The concentration of histamine in the prenatal brain is fivefold that of adult levels. During the prenatal development a large transiently histamine-immunoreactive cell population distinct from the adult histaminergic system can be found within a subpopulation of the developing serotonergic raphe nuclei neurons. Also histamine-immunoreactive nerve fibers are widely distributed already during the prenatal development extending to the diencephalon, the thalamus, the cortex, and the spinal cord. Large numbers of histamine-containing mast cells also migrate into the brain during the late prenatal life. The wide distribution and high prenatal concentrations imply important functions for the histaminergic system during intrauterine development. However, little is known about the actual functions of histamine during development, and which of the histamine receptors are present in the prenatal rat brain is currently unknown. In the present study, we used in situ hybridization to study the distribution of H1-receptor (H1R) mRNA in the embryonic rat brain and spinal cord. H1R mRNA could be detected in rat brain and in spinal cord on embryonic day (E) 14, and the expression pattern seemed to partially localize in areas containing histamine-immunoreactive nerve fibers through E14-E20. H1R mRNA was also detected by reverse transcriptase polymerase chain reaction from embryonic brain samples and by Northern hybridization. The possible involvement of apoptosis in the disappearance of the developing transiently histaminergic system was studied by using apoptosis detection based on the terminal dUTP nick end labeling (TUNEL) technique and with c-Fos immunostaining. Although histamine immunoreactivity disappears dramatically from the developing raphe nuclei after E18, only occasional apoptotic nuclei could be seen in the histamine-immunoreactive cell bodies. The presence of H1R mRNA during the embryonic development renders it possible that histamine could exert an H1R-specific function at the time of the embryonic histamine peak.


Unique binding pocket for KW-4679 in the histamine H1 receptor

The histamine H1 receptor has an aspartate (Asp) residue in transmembrane helix 3 (TM3), which is well-conserved among biogenic amine receptors. The Asp residue is one of the most crucial amino acids for ligand binding. The tested histamine H1 receptor antagonists with tri- and tetracyclic structures were not selective for histamine H1 receptors and showed affinity for several other biogenic amine receptors. In contrast, KW-4679 ((Z)-11-[3-(dimethylamino)propylidene]-6,11-dihydrodibenz[b, e]oxepin-2-acetic acid hydrochloride), a tricyclic compound, was a selective histamine H1 receptor antagonist. [3H]KW-4679 had high affinity (Kd value of 2.5 +/- 0.12 nM) for wild-type human histamine H1 receptors. In the [3H]KW-4679 binding assay, replacement of Asp107 by alanine by site-directed mutagenesis greatly reduced the affinities (280-2100-fold) of tri- and tetracyclic compounds, whereas this mutation led to a comparatively small reduction (14-fold) in KW-4679 affinity. These results demonstrate that the tested tri- and tetracyclic histamine H1 receptor antagonists which have a tight interaction with the Asp residue are not selective for the histamine H1 receptor. Furthermore, the high selectivity of KW-4679 might be explained by a unique binding pocket, which consists of the Asp residue and other acceptor sites, in the histamine H1 receptor.


Histamine-induced calcium mobilization in single cultured cells expressing histamine H1 receptors: a relationship between its sensitivity and the density of H1 receptors

International Journal of Molecular Medicine


Regulation of the human histamine H1 receptor stably expressed in Chinese hamster ovary cells


Plasma diamine oxidase activity in asthmatic children


Mobilization of gastric histamine during repeated administration of a proton potassium adenosine triphosphatase inhibitor in intact and antrectomized rats

Intact and antrectomized female rats were treated with the potent proton pump inhibitor, E3810 (daily 40 mg/kg weight, s.c.) for 4 weeks. Plasma gastrin concentration and urinary excretion of N-terminal big gastrin increased until day 14 and persisted at a high level in intact rats treated with E3810, but did not increase in antrectomized rats. Urinary excretion of histamine increased progressively and reached 7 times the control value following 4 weeks of treatment with E3810 in intact rats, but not in antrectomized rats. At the termination of the treatment, the endocrine cell density in the oxyntic mucosa of intact rats had increased by 85% with increased histamine content and elevated histidine decarboxylase activity, while antrectomized rats showed a low histamine level and low histidine decarboxylase activity. Administration of gastrin-17 I (10 micrograms/kg weight, sc) itself caused a significant increase in urinary excretion of histamine, which was inhibited by the specific gastrin receptor antagonist, L-365,260. These results suggests that the massive urinary excretion of histamine caused by the treatment with E3810 reflects gastrin-induced mobilization of gastric histamine and that neither E3810 itself nor E3810-induced luminal pH elevation has direct effects on mobilization of oxyntic mucosal histamine.


Helicobacter pylori Increases Gene Expression of Hepatocyte Growth Factor in Human Gastric Mucosa

Biochemical and Biophysical Research Communications


Purification and characterization of [3H]mepyramine (histamine H1 antagonist)-binding protein from rat liver: a highly homologous protein with cytochrome P450 2D


Determination of histidine decarboxylase mRNA in various rat tissues by the polymerase chain reaction


Mepyramine, a histamine H1 receptor antagonist, inhibits the metabolic activity of rat and human P450 2D forms

The Journal of Pharmacology and Experimental Therapeutics


Purification and characterization of [3H]mepyramine (histamine H1 antagonist)-binding protein from rat liver: a highly homologous protein with cytochrome P450 2D.


Formation of inositol phosphates mediated by M3 muscarinic receptors in type-1 and type-2 astrocytes from neonatal rat cerebral cortex.

Muscarinic receptor subtype in type-1 and type-2 astrocytes from rat neonalal cerebral cortex was examined for carbachol-elicited inositol phosphate (IP) formation. The formation of carbachol-elicited IP was inhibited by various muscarinic antagonists in the following relative order of potency: 4-DAMP > or = atropine > pirenzepine > AF-DX 116. This pharmacological profile suggests that the activation of the M3 muscarinic receptor subtype is responsible for the stimulation of IP formation in both astrocytes.


Expression of the histamine H1 receptor gene in relation to atherosclerosis


Interleukin-1β inhibits gastric histamine secretion and synthesis in the rat.


Glucocorticoids enhance histamine-evoked catecholamine secretion from bovine chromaffin cells.


Does [3H]mepyramine binding site represent the histamine H1 receptor? Re-examination of the histamine H1 receptor with quinine.

Journal of Pharmacology et Experimental Therapeutics


Site-directed mutagenesis of the histamine H1 receptor: roles of aspartic acid107, asparagine198 and threonine194.


Purification and characterization of diamine oxidase (histaminase) from rat small intestine.


Molecular cloning of the human histamine H1 receptor gene.


Expression and characterization of the bovine histamine H1 receptor in cDNA-transfected C6 astroglioma cells.


Tissue distribution of histamine N-methyltransferase-like immunoreactivity in rodents.


Role of nucleosides and nucleotide mixture in intestinal mucosal growth under total parenteral nutrition.

The preventive effect of a mixture of nucleosides and a nucleotide (OG-VI) on total parenteral nutrition (TPN)-associated gut mucosal atrophy was investigated. Male Wistar rats were randomly divided into two groups. The TPN group (n = 11) received a standard TPN diet (250 kcal and 1.78 g nitrogen.kg-1.day-1) for 6 days; the TPN + OG-VI group (n = 10) received OG-VI (2.5 ml.kg-1.day-1) in addition to the standard TPN solution for 6 days. To obtain information on a normal fed condition of the intestine, another 10 rats were maintained on oral rat chow for 6 days. Compared with the TPN group, mucosal wet weights in the jejunum (22.8 +/- 1.8 vs. 20.5 +/- 2.0 mg/cm) and the ileum (19.4 +/- 2.2 vs. 16.8 +/- 2.3 mg/cm) were significantly greater in the TPN + OG-VI group. Likewise, diamine oxidase activities in the jejunum (13.7 +/- 4.27 vs. 8.02 +/- 3.40 nmol.min-1.cm-1) and the ileum (33.9 +/- 6.89 vs. 25.9 +/- 7.93 nmol.min-1.cm-1) were significantly higher in the TPN + OG-VI group. Moreover, the bromodeoxyuridine labeling index in the TPN + OG-VI group (36.9 +/- 4.30%) was significantly higher than in the TPN group (31.0 +/- 3.03%). The addition of OG-VI to the standard TPN diet improved mucosal growth and maturity by increasing the proliferating activity of crypt cells. External provision of purines and pyrimidines may be necessary to sustain mucosal function during TPN.


High stable expression of the bovine histamine H1-receptor coupled to [Ca2+]i mobilization in CHO-K1 cells.

The bovine H1-receptor DNA was transfected into Chinese hamster ovary cells (CHO-K1) using an expression vector. Binding studies revealed very high expression levels of the receptor which was found to have a Kd for [3H]-mepyramine of 1 nM. Addition of histamine resulted in a concentration-dependent increase in intracellular calcium which was found to involve both release from intracellular stores and entry across the plasma membrane. Furthermore, the response demonstrated the pharmacological characteristics of an H1-receptor-mediated event. Thus, we present the first report of the high, functional expression of the bovine H1-receptor coupled to mobilisation of intracellular calcium in CHO-K1 cells.

Biochemical and Biophysical Research Communications


Molecular cloning of guinea-pig histamine H1 receptor gene.

The histamine H1 receptor gene was isolated from a guinea-pig gene library. The gene contains no introns and encodes a protein of 488 amino acid residues. The structure of the guinea-pig histamine H1 receptor is predicted to contain seven putative transmembrane regions, which are similar to those of receptors coupling with GTP binding proteins. Although the third intracellular domain, the predicted binding site for the GTP binding protein, showed only 50% identity with those of the bovine and rat H1 receptors, the expressed guinea-pig H1 receptor was fully able to bind with [3H]mepyramine. Northern blot analysis indicated that the cerebrum, cerebellum, lung, adrenal, intestine, and heart expressed 3.3 kb guinea-pig H1 receptor mRNA. Expression of histamine H1 mRNA of guinea-pig peripheral organs was greater than that of rat organs, suggesting the high sensitivity of guinea-pig organs as to histamine is due to the contents of histamine H1 receptor mRNA. In addition, the lung, adrenal, intestine, and heart expressed 3.9 kb mRNA. In situ hybridization showed that the hippocampus, cerebral cortex, thalamus, and granular layer of the cerebellum each contained a large amount of histamine H1 receptors. Southern blot analysis showed that there was another gene quite similar to the cloned histamine H1 receptor gene.


Genomic cloning of the rat histamine H1 receptor.

A rat histamine H1 receptor gene which lacked introns was isolated from a rat genomic library using recently cloned bovine histamine H1 receptor cDNA [Yamashita et al., Proc. Natl. Acad. Sci. USA, 88, 11515-11519 (1991)]. The receptor protein deduced from this isolated gene was composed of 486 amino acids and showed characteristic properties of G protein-coupled receptors. At the 5'-flanking region of the receptor gene, we have located potential TATA box sequences and consensus sequences for the glucocorticoid response element and AP-2 element. After being subcloned into a mammalian expression vector, the isolated gene was transfected to C6 glioma cells. These cells showed significant binding toward [3H]mepyramine. The binding was inhibited by H1 antagonists or histamine. The mode of this binding was comparable to the binding of membranes derived from rat tissues toward [3H]mepyramine. Northern blot analysis detected a 3.0 kb nucleotide band for histamine H1 receptor mRNAs from rat brain and small intestine when these mRNAs were hybridized with the isolated rat H1 gene. The present results demonstrate the isolation of the rat histamine H1 receptor gene.


Histamine content, synthesis and degradation in human nasal mucosa.


Histamine content, synthesis and degradation on nasal mucosa and lung of guinea-pig treated with toluene diisocyanate (TDI).


Somatostatin inhibits gastrin-induced histamine secretion and synthesis in the rat.


Vasopressin-induced changes in brain histamine and H-1 receptors.


Effect of capsaicin desensitization on nasal allergy-like symptoms and histamine release in the nose induced by toluene diisocyanate in guinea pigs.


Type-1 and type-2 astrocytes are distinct targets for prostaglandins D2, E2 and F2α.

Accumulating evidence has revealed that astrocytes are potential targets for various neurotransmitters. Here we investigated the effects of prostaglandins (PGs) on signal transduction in purified primary cultures of rat type-1 and type-2 astrocytes. PGF2 alpha, PGD2, and 9 alpha,11 beta-PGF2, a metabolite of PGD2 and a stereoisomer of PGF2 alpha, evoked a rapid rise in the intracellular Ca2+ concentration ([Ca2+]i) in type-1, but not in type-2, astrocytes. STA2, a stable analogue of thromboxane A2, was less effective, and PGE2 showed little effect. The PG-induced rise in [Ca2+]i was not blocked by an antagonist of either PGD2 receptor or thromboxane A2 receptor. PGF2 alpha and 9 alpha,11 beta-PGF2 stimulated rapid formation of inositol trisphosphate followed by inositol bisphosphate and inositol monophosphate. On the other hand, PGE2 increased the intracellular level of cyclic AMP in type-2 astrocytes, rather than in type-1 astrocytes. The potency of PGs for cyclic AMP formation was in the following order: PGE2 greater than PGE1 greater than or equal to STA2 much greater than iloprost, a stable analogue of PGI2. PGD2 and PGF2 alpha had no effect on cyclic AMP formation. These results demonstrate that type-1 astrocytes preferentially express PGF2 alpha receptors, the activation of which leads to phosphoinositide metabolism and [Ca2+]i elevation, whereas type-2 astrocytes possess PGE receptors that are linked to cyclic AMP formation.


Re-examination of [3H]mepyramine binding assay for histamine H1 receptor using quinine.


Histamine N-methyltransferase from rat kidney. Cloning, nucleotide sequence, and expression in Escherichia coli cells.


Expression cloning of a cDNA encoding the bovine histamine H1 receptor.

Proceedings of the National Academy of Sciences of the United States of America


Adenosine induces System A amino acid transport in cultured rat hepatocytes.


Presynaptic localization of histamine H3-receptors in rat brain.

Biochemical and Biophysical Research Communications


Synaptic and extra-synaptic distribution of histamine H1-receptors in rat and guinea pig brains.


Immuno-cross-reactivity of histidine and dopa decarboxylases.


ナノ粒子による炎症性伝達物質吸着を利用した炎症制御の研究

ナノ粒子による炎症性伝達物質吸着を利用した炎症制御の研究

ケミカルエンジニヤリング


Up-regulation of histamine H1 receptors in an allergic rat nasal mucosa model


Increase in the level of histidine decarboxylase mRNA expression in nasal mucosa of rats sensitized by toluene diisocyanate