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N Grundström

Publications and source records attributed to N Grundström.

At least 19 recordsLinked to original sources
PubMed1997

Multiple alpha 2-adrenoceptor signalling pathways mediate pigment aggregation within melanophores.

It has previously been shown that alpha 2-adrenoceptors (alpha 2-ARs) mediate pigment granule (melanosome) aggregation in melanophores of the teleost fish Labrus ossifagus. The present investigation scrutinized the signalling mechanisms of melanosome aggregation induced by sympathetic nerve stimulation or by exogenous addition of alpha-AR agonists and cAMP analogues. The following was observed: i) nerve-induced melanosome aggregation was associated with a rapid decrease in the cAMP level; ii) noradrenaline or medetomidine (an alpha 2-AR agonist) caused melanosome aggregation and reduced the cAMP content; iii) RP-8-Cl-cAMP, a membrane-permeating inhibitor of protein kinase A induced melanosome aggregation; and iv) B-HT 920 (an alpha 2-AR agonist) and methoxamine (an alpha 1-AR agonist) induced melanosome aggregation, although they did not reduce cAMP. It has been suggested that in some teleost species alpha 1-ARs mediate melanosome aggregation by increasing the level of intracellular calcium. However, we found that the effect of methoxamine in melanophores from Labrus ossifagus could be blocked by yohimbine (an alpha 2-AR antagonist) but not by equimolar concentration of prazosin (an alpha 1-AR antagonist). Furthermore, 1 microM ionomycin (a calcium ionophore) did not induce melanosome aggregation. Our findings therefore do not indicate that alpha 1-ARs and/or an increase in intracellular calcium mediate melanosome aggregation in Labrus ossifagus. Our results suggest that alpha 2-AR-mediated melanosome aggregation is induced by multiple signalling pathways. One of these involves a reduction in cAMP, but none involves an increase in intracellular calcium.

Animals↗
PubMed1995

Pertussis toxin attenuates presynaptic inhibition of cholinergic but not nonadrenergic noncholinergic contraction in guinea pig airways.

Several alpha-2- and opioid receptors exert their intracellular effects via a pertussis toxin-sensitive G protein. Both cholinergic and nonadrenergic noncholinergic nerve-mediated contractions of airways can be reduced by stimulation of presynaptic alpha-2- and opioid receptors. Using guinea pig trachea and bronchi, pretreated with pertussis toxin 100 micrograms/kg or 0.9% NaCl, we investigated whether these inhibitory effects on airway contractions, evoked by electrical field stimulation, are mediated via a pertussis toxin-sensitive G protein. The results indicate a difference between cholinergic and excitatory nonadrenergic non-cholinergic neurotransmission. Inhibition of cholinergic contraction via presynaptic alpha-2- and opioid receptors seems to be mediated via a pertussis toxin-sensitive G protein, whereas inhibition of nonadrenergic noncholinergic contraction is not affected by pertussis toxin.

Animals↗
PubMed1993

Cloning and expression of a fish alpha 2-adrenoceptor.

1. Pigment granule aggregation in specialized cells (melanophores) from the skin of teleost fishes has been shown to be mediated by receptors with an alpha 2-adrenoceptor pharmacology. We now report the cloning of the alpha 2-F, a fish skin alpha 2-receptor from the cuckoo wrasse (Labrus ossifagus). 2. Degenerate oligonucleotides corresponding to conserved regions of the human alpha 2-adrenoceptor subtypes were used in a polymerase chain reaction (PCR) with cDNA prepared from mRNA isolated subtypes were used in a polymerase chain reaction (PCR) with cDNA prepared from mRNA isolated from the skin of the cuckoo wrasse. An 876 base pair (bp) product was obtained that was homologous with that of the human alpha 2-adrenoceptor and was used to screen a genomic library from the cuckoo wrasse. 3. A clone (pTB17BS) consisting of approximately 5 kb of genomic DNA was obtained which contained the nucleotide sequence of the initial PCR product. In addition, it contained an open reading frame that encoded a protein of 432 amino acids and approximately 2 kb of 5'untranslated sequence. The deduced amino acid sequence of this protein showed 47-57% identity with the human alpha 2-adrenoceptors and thus appeared to encode a fish alpha 2-adrenoceptor. 4. In the 5'-untranslated region of the gene, nucleotide sequences were present suggesting that transcription of the alpha 2-F might be regulated by cyclic AMP, calcium and/or steroids. 5. The alpha 2-F was expressed in COS-7 cells and radioligand binding studies were performed with [3H]-rauwolscine. The binding was of high affinity and it was saturable with a KD of 0.8 +/- 0.1 nM and a Bmax of 5.7 +/- 1.0 pmol mg-1 of protein.6. Competition curves for the displacement of specific [3H]-rauwolscine binding showed the following order of potency: for agonists, medetomidine > clonidine >p-aminoclonidine> B-HT 920> (- )-noradrenaline;for antagonists, rauwolscine > atipamezole > yohimbine > phentolamine > prazosin.7. These results show that alpha2-F has characteristics of both the human alpha2-CIO and alpha2-C4 and that it might represent an ancestral alpha2-adrenoceptor subtype.

Amino Acid Sequence↗
PubMed1993

Involvement of nitric oxide and peptides in the inhibitory non-adrenergic, non-cholinergic (NANC) response in bovine mesenteric artery.

The cyclic GMP mediated non-adrenergic non-cholinergic (NANC) relaxation in field stimulated bovine mesenteric artery and its modulation by various factors was studied. Electrical field stimulation of precontracted (Phe 2.5 microM or histamine 5 microM) bovine mesenteric arteries resulted in relaxations varying between 10-70% in different preparations. Tetrodotoxin (3 microM) completely blocked the inhibitory NANC response. Preincubation with high concentrations (100 microM-1 mM) of NG-nitro-L-arginine for 15 min. significantly reduced the relaxation induced by electrical field stimulation. Blockade of cyclooxygenases and prostaglandin synthesis by indomethacin had no effect on the relaxatory response to electrical field stimulation. Neither the alpha 2-adrenoceptor selective antagonist yohimbine (1 microM) nor the alpha 2-adrenoceptor selective agonist UK 14,304 (1 microM) had any significant effect on the electrical field stimulation-induced relaxation. Pertussis toxin (100 ng/ml) was without effect on relaxations elicited by electrical field stimulation. GTP in the concentration range 10 microM-1 mM slightly potentiated the relaxant response. N-carboxymethyl-Phe-Leu (an inhibitor of enkephalinase) or aprotinin (an inhibitor of several proteases) had no significant effect on the electrical field stimulation response. Addition of trypsin (100 U/ml) in combination with chymotrypsin (20 U/ml) significantly reduced the electrical field stimulation-induced relaxation. In the present study we have found indications for the involvement of nitric oxide and possibly also peptides in mediating the inhibitory NANC response (relaxation) in bovine mesenteric arteries.

Adrenergic alpha-Agonists↗
PubMed1992

Relationship between histamine, lipoxygenase and cyclooxygenase products in antigen-induced contraction in guinea-pig tracheal tube preparations.

We have used a tracheal tube preparation to study antigen-induced contraction in sensitized guinea pig airways. Treatment with both the cyclooxygenase inhibitor indomethacin and the lipoxygenase inhibitor MK-886 (L-663,536) affected this contraction in preparations with intact epithelium. Indomethacin potentiated and MK-886 inhibited part of the contraction. Leukotriene release from tracheal tubes was measured after antigen challenge, and was found to be significant in preparations with an intact epithelium. When the epithelium was removed, the histamine receptor antagonist mepyramine reduced antigen-induced contraction by 90%. Our results show that when the epithelium is absent, histamine is the most important mediator in the contraction. With the epithelium left intact, the contraction is more complex: both the cyclooxygenase and lipoxygenase pathways are involved, and our findings indicate that eicosanoid production is associated with the airway epithelium.

Animals↗
PubMed1992

The modulatory effects of prostaglandins on both excitatory and inhibitory non-adrenergic non-cholinergic neurotransmission in guinea-pig airways.

Guinea-pig trachea and bronchi were used to investigate the effects of indomethacin and prostaglandin E2 (PGE2) on non-adrenergic non-cholinergic excitatory (e-NANC) and inhibitory (i-NANC) neurotransmission evoked by electrical field stimulation. Indomethacin potentiated e-NANC responses in bronchi with intact epithelium but had no effect on epithelium-denuded preparations. Inhibitory NANC responses were increased by indomethacin independent of the epithelium. Both i-NANC and e-NANC neurotransmission were suppressed by PGE2 in a dose-dependent manner. These results indicate that endogenous prostaglandins (e.g. PGE2) generated from the epithelium have an inhibitory effect on i-NANC and e-NANC nerve responses in airways. The epithelium is presumably not the only source for generation of prostaglandins that are involved in i-NANC neurotransmission.

Acetylcholine↗
PubMed1992

Epithelial modulation of allergen and drug effects in guinea pig airways.

The effect of egg albumin (EA) challenge on tracheal tube preparations from sensitized guinea pigs was studied with regard to EA permeability, histamine release and penetrability, and the contractile response of the preparation. We used a plethysmographic method that allowed simultaneous measurement of changes in smooth muscle tension and collection of samples for determination of mediators. Our results clearly show that epithelial damage potentiates the contractile response to histamine, potassium ions, and acetylcholine. Epithelial damage did not alter the maximal contractile response in preparations challenged with high antigen concentrations (EA, 1 mg/ml), but histamine release measured in the perfusate increased substantially. The permeability of the preparations to EA was greater when the epithelium was damaged. No increase in the permeability in response to the EA challenge was observed. The present study has demonstrated that guinea pig airway epithelium constitutes a barrier for both antigen and drugs. We also present a method for recording contractile responses from intact whole tracheal preparations, in which the epithelium can still act as a barrier, as is the case in vivo.

Allergens↗
PubMed1991

The melanophore aggregating response of isolated fish scales: a very rapid and sensitive diagnosis of whooping cough.

Pertussis toxin (PT) has been found to block noradrenaline-induced pigment aggregation in fish melanophores, and, based on this, a rapid and highly sensitive assay for PT was developed. Some preliminary results have also indicated that it may be possible to detect PT-like activity in saliva samples from patients with clinically suspected pertussis. In the present study the diagnostic value of the fish melanophore method was evaluated in 70 patients suspected of having pertussis; culture, serology and physician diagnosis were used as reference methods. In 60 of the patients, pertussis was verified by at least one of the reference methods. The melanophore test showed PT-like activity in saliva samples from 58 of the patients. Three patients with reference-verified pertussis showed no PT-like activity in the test; among these, one patient had been immunized and had also been treated with erythromycin during 3 days immediately prior to visiting the hospital. The melanophore test has three major advantages: it allows detection of pertussis in the early and curable stage of the disease; it takes only 2 h to perform; and it requires no sophisticated equipment.

Adolescent↗
PubMed1991

Is the airway epithelium responsible for histamine metabolism in the trachea of guinea pigs?

The aim of the study was to investigate whether or not the epithelium plays an active role in histamine metabolism, via the histaminase and/or methyltransferase pathways. Isolated tracheas from guinea pigs sensitized to egg albumin (EA) were used. The epithelium was either left intact or removed from the preparations. The tracheal tubes were mounted in a chamber, allowing estimation of smooth muscle tension, and perfused with buffer. In some experiments the perfusate was collected for determination of histamine and methylhistamine. Mepyramine was used to evaluate the contribution of histamine to the EA-induced contraction. Mepyramine reduced the contraction by 90% when the epithelium was removed; with intact epithelium the reduction was 47%. Aminoguanidine, a histaminase inhibitor, significantly potentiated the response to EA when the epithelium was left intact. Traces of methylhistamine were detected in tissue extracts and perfusates. We conclude that histaminase is present in the preparation and that it can contribute to the inhibitory effect of the epithelium by means of histamine degradation.

Amine Oxidase (Copper-Containing)↗
PubMed1991

MCH-induced pigment aggregation in teleost melanophores is associated with a cAMP reduction.

It has previously been shown that alpha 2-adrenoceptors are involved in noradrenaline-induced pigment aggregation within fish melanophores. In the present investigation, melanin concentrating hormone (MCH) elicited pigment aggregation (EC50 approximately 1 x 10(-7) M) that was associated with a significant reduction in the cAMP content; 1 x 10(-7) M MCH reduced the cAMP content from a basal level of 50.4 +/- 2.8 pmol/mg protein to 36.9 +/- 3.8 pmol/mg protein. Like the alpha 2-adrenoceptor-induced pigment aggregation, the MCH response was effectively blocked by the adenylate cyclase stimulator forskolin. These findings suggest that attenuation of cAMP may serve as an intracellular signal transduction mechanism for both MCH and noradrenaline.

Animals↗
PubMed1991

Influence of some alpha 2-receptor agonists and antagonists on the excitatory non-adrenergic, non-cholinergic neurotransmission in the airways of guinea-pigs in vivo.

Neural control of smooth muscle tension in the airways of guinea-pigs can be subdivided into at least four components: an excitatory cholinergic, an inhibitory noradrenergic, an excitatory non-adrenergic, non-cholinergic and an inhibitory non-adrenergic, non-cholinergic component. The existence of alpha 2-adrenoceptors that modulate the excitatory non-adrenergic, non-cholinergic nerve activity has also been demonstrated. The aim of the present study was to investigate the dose-dependence of the selective alpha 2-receptor agonists UK 14,304, dexmedetomidine and clonidine on these alpha 2-adrenoceptors in the airway of guinea-pigs in vivo. Electrical stimulation of the cervical vagus nerves in anaesthetized guinea-pigs resulted in a biphasic response in insufflation pressure: an immediate increase due to excitation of cholinergic nerve fibres and a slower, longer-lasting increase due to activation of the excitatory non-adrenergic, non-cholinergic fibres to be studied. The cholinergic compound was abolished by atropine, leaving the excitatory non-adrenergic, non-cholinergic component as the sole response recorded during vagal stimulation. UK 14,304 and dexmedetomidine attenuated the response in a dose-dependent manner, whereas the effect of the partial agonist clonidine was less dose-related. UK 14,304 and dexmedetomidine reduced the insufflation pressure about 60 and 53% respectively at the highest dose. In conclusion, the neurotransmission in the excitatory non-adrenergic, non-cholinergic nerves is attenuated by selective alpha 2-receptors in a dose-dependent manner. The present study adds support to earlier observations that the excitatory non-adrenergic, non-cholinergic neural system in the airways of guinea-pigs is modulated by alpha 2-adrenoceptors.

Adrenergic alpha-Agonists↗
PubMed1991

Role of nitric oxide and cyclic GMP as mediators of endothelium-independent neurogenic relaxation in bovine mesenteric artery.

Electrical field stimulation (EFS) of phenylephrine-contracted bovine mesenteric arteries pretreated with guanethidine elicited a relaxation that amounted to roughly 40%. This relaxation was sensitive to tetrodotoxin pretreatment, suggesting a neurogenic origin. The EFS-induced relaxation was correlated to an increase in cGMP level, from 14.2 +/- 2.5 pmol/g wet wt in nonstimulated arteries to 31.6 +/- 3.4 pmol/g wet wt after 1 minute of EFS. cAMP values were not affected by EFS. Methylene blue (5 microM) and the compound LY 83583 (10 microM), inhibitors of soluble guanylate cyclase, inhibited the EFS-induced relaxation by 60% and 50%, respectively. Zaprinast (1 microM), a selective inhibitor of cGMP degradation, significantly (p = 0.005) potentiated the EFS-induced relaxation. The relaxation induced by EFS in bovine mesenteric arteries exhibits characteristics similar to the relaxations evoked by organic nitroesters and endothelium-dependent vasodilators, both of which are suggested to be mediated by cGMP and probably with nitric oxide as the common activator of the cGMP system. The possible involvement of nitric oxide as a mediator of EFS-induced relaxations was investigated with the use of known modulators of endogenous nitric oxide production. Preincubation of the arteries with 1 mM arginine or 1 mM N-alpha-benzoyl-L-arginine, both reported to potentiate endogenous nitric oxide production, or 5 mM L-canavanine, 0.25 mM NG-monomethyl-L-arginine, or 0.1 mM NG-nitro-L-arginine, alleged inhibitors of endogenous nitric oxide production, were without effect on the relaxation induced by EFS. However, pyrogallol, a generator of superoxide anions, was a potent inhibitor of relaxations induced by EFS in bovine mesenteric arteries.(ABSTRACT TRUNCATED AT 250 WORDS)

3',5'-Cyclic-GMP Phosphodiesterases↗
PubMed1991

Antagonistic effect of amiloride on alpha-2 adrenoceptor-mediated pigment aggregation: pharmacological heterogeneity between B-HT 920 and noradrenaline.

Pigment aggregation in melanophores from the cuckoo wrasse (Labrus ossifagus L.) has previously been shown to be mediated by alpha-2 adrenoceptors. In the present investigation, the effect of amiloride on pigment aggregation induced by noradrenaline and B-HT 920 was studied. Amiloride caused a parallel shift to the right in concentration-response curves for 5-allyl-2-amino-5,6,7,8-tetrahydrothiazolo-[4,5-d]azepine-dihyd roc hloride: (B-HT 920), with no change in the maximal response, indicating simple competitive inhibition. Subsequent Schild plot analysis gave a straight line with a slope of almost unity (0.95) and a KB of 5 x 10(-6) M. In contrast to the effect of amiloride on B-HT 920-induced pigment aggregation, only a very high concentration of amiloride (1 x 10(3) M) inhibited the corresponding effect of noradrenaline. Na+/H+ exchange has been suggested as a possible mechanism for alpha-2 adrenoceptors, and the amiloride analogs 5-(N,N-dimethyl)amiloride and 5-(N,N-hexamethylene)amiloride are known to be more specific than amiloride itself in this respect. However, these analogs inhibited the effect of B-HT 920 significantly less effectively than amiloride. Furthermore, manipulating the extracellular pH between 6.8 and 7.8 did not affect the concentration response curves of B-HT 920. Neither did the prostanoid pathway inhibitors, quinacrine or indomethacin, inhibit the pigment-aggregating effect of B-HT 920. The present results suggest that amiloride competes with B-HT 920 as receptor antagonist, and that Na+/H+ exchange is insignificant for the alpha-2 adrenoceptor-stimulated pigment aggregation.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenergic alpha-Agonists↗
PubMed1990

Can the epithelium affect allergen-evoked histamine release from the perfused guinea pig trachea?

The aim of this study was to investigate the influence of the epithelium on allergen-evoked histamine release. Isolated tracheal preparations from guinea-pigs, with the epithelium either left intact or removed, were used. The trachea was perfused and challenged with egg-albumin (EA) either extra- or intraluminally. Fractions of the perfusate were collected for analysis of the histamine and EA contents. When EA was added intraluminally and the epithelium was removed, a marked peak in histamine release could be detected. In the presence of intact epithelium there was no marked histamine peak. EA added extraluminally gave the same results but the histamine peak was less pronounced. The epithelium apparently acts as a barrier, the nature of which remains to be elucidated.

Allergens↗
PubMed1990

Fish scales as biosensors for catecholamines.

Certain fish scales contain specialized cells (chromatophores) with pigment granulas, which can be dispersed or aggregated in the cells. The degree of aggregation is determined by a transmitter substance, noradrenaline, released by the sympathetic nerve endings in the skin. Isolated scales from, for example, cuckoo wrasse (Labrus ossifagus) retain a large sensitivity to externally applied noradrenaline (or more generally catecholamines) for several weeks. The degree of aggregation in isolated scales can be measured objectively by simple photometric techniques. We demonstrate in this paper how fish scales can be used to monitor catecholamine levels in human blood plasma. A discussion of other potential biosensor applications of this intact biological receptor-effector system is also given.

Animals↗
PubMed1990

Suppression by neuropeptide Y of capsaicin-sensitive sensory nerve-mediated contraction in guinea-pig airways.

1. In the present study we have examined whether neuropeptide Y (NPY) interferes with non-adrenergic, non-cholinergic nerve-mediated contractions and relaxations in the guinea-pig airways. In these experiments we have used ring preparations of bronchi and trachea, incubated in the presence of atropine, propranolol and indomethacin (each 1 microM). 2. The contractile response to electrical stimulation of non-adrenergic, non-cholinergic nerve fibres was suppressed by NPY and NPY 13-36 in a concentration-dependent manner, these agents having similar inhibitory potencies. NPY caused a more complete inhibition than the C terminal fragment. 3. NPY affected neither the basal tension nor the substance P-evoked contraction in the bronchi and trachea and did not interfere with nerve-mediated, non-adrenergic relaxation in the trachea. 4. On the basis of these results, it is suggested that NPY may act on the terminals of sensory neurones in the airways to prevent antidromic, excitatory neurotransmission by inhibiting transmitter release.

Animals↗