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Biomedical subjects

R P Markus

Publications and source records attributed to R P Markus.

At least 19 recordsLinked to original sources
PubMed2002

Evaluation of the cholinomimetic actions of trimethylsulfonium, a compound present in the midgut gland of the sea hare Aplysia brasiliana (Gastropoda, Opisthobranchia).

Trimethylsulfonium, a compound present in the midgut gland of the sea hare Aplysia brasiliana, negatively modulates vagal response, indicating a probable ability to inhibit cholinergic responses. In the present study, the pharmacological profile of trimethylsulfonium was characterized on muscarinic and nicotinic acetylcholine receptors. In rat jejunum the contractile response induced by trimethylsulfonium (pD2 = 2.46 +/- 0.12 and maximal response = 2.14 +/- 0.32 g) was not antagonized competitively by atropine. The maximal response (Emax) to trimethylsulfonium was diminished in the presence of increasing doses of atropine (P<0.05), suggesting that trimethylsulfonium-induced contraction was not related to muscarinic stimulation, but might be caused by acetylcholine release due to presynaptic stimulation. Trimethylsulfonium displaced [3H]-quinuclidinyl benzilate from rat cortex membranes with a low affinity (Ki = 0.5 mM). Furthermore, it caused contraction of frog rectus abdominis muscles (pD2 = 2.70 +/- 0.06 and Emax = 4.16 +/- 0.9 g), which was competitively antagonized by d-tubocurarine (1, 3 or 10 microM) with a pA2 of 5.79, suggesting a positive interaction with nicotinic receptors. In fact, trimethylsulfonium displaced [3H]-nicotine from rat diaphragm muscle membranes with a Ki of 27.1 microM. These results suggest that trimethylsulfonium acts as an agonist on nicotinic receptors, and thus contracts frog skeletal rectus abdominis muscle and rat jejunum smooth muscle via stimulation of postjunctional and neuronal prejunctional nicotinic cholinoreceptors, respectively.

Animals↗
PubMed2001

Melatonin and N-acetylserotonin inhibit leukocyte rolling and adhesion to rat microcirculation.

The hormone melatonin produced by the pineal gland during the daily dark phase regulates a variety of biological processes in mammals. The aim of this study was to determine the effect of melatonin and its precursor N-acetylserotonin on the microcirculation during acute inflammation. Arteriolar diameter, blood flow rate, leukocyte rolling and adhesion were measured in the rat microcirculation in situ by intravital microscopy. Melatonin alone or together with noradrenaline did not affect the arteriolar diameter or blood flow rate. Melatonin inhibited both leukocyte rolling and leukotriene B(4) induced adhesion while its precursor N-acetylserotonin inhibits only leukocyte adhesion. The rank order of potency of agonists and antagonist receptor selective ligands suggested that the activation of MT(2) and MT(3) melatonin binding sites receptors modulate leukocyte rolling and adhesion, respectively. The effect of melatonin and N-acetylserotonin herein described were observed with concentrations in the range of the nocturnal surge, providing the first evidence for a possible physiological role of these hormones in acute inflammation.

Animals↗
PubMed2001

Interaction between the adrenal and the pineal gland in chronic experimental inflammation induced by BCG in mice.

OBJECTIVE: To investigate the adrenal gland influence on diurnal rhythm of chronic inflammation induced by BCG in mice and its interaction with the pineal gland. METHODS: C57Bl/6 mice were injected with BCG in the footpad and maintained in a 12/12 h light-dark cycle. All the experimental manipulations were done after 20-45 days. Paw swelling was measured every 4 h for 48 or 72 h and decomposed by Fourier transformation. Vascular permeability was evaluated by Evans Blue overflow, in mice killed at midday or midnight. 6-Sulphatoxymelatonin urine concentration was determined by radioimmunoassay in samples taken during the dark or light phase. RESULTS: Adrenalectomy or metyrapone treatment abolished the paw swelling diurnal rhythm, the nocturnal reduction in vascular permeability, and the nocturnal increase in 6-sulphatoximelatonin in the urine. Nocturnal administration of melatonin to adrenalectomized mice restored the paw swelling diurnal variation and the reduction of vascular permeability of the inflamed paw. CONCLUSION: Adrenal cortical hormones are important for the maintenance of the diurnal rhythm of chronic inflammation (paw swelling and vascular permeability), probably by promoting a nocturnal surge of melatonin, which is the hormone that modulates the diurnal variation of chronic inflammation.

Adrenal Glands↗
PubMed2001

Characterisation of P2Y(1)-like receptor in cultured rat pineal glands.

The rat pineal gland possesses P2 receptors which potentiate the effect of noradrenaline-induced N'-acetyl-5-hydroxytryptamine (N'-acetyl-5-HT) production. In the current study, this receptor was characterised according to agonist selectivity and signal transduction mechanisms. 2-MethylthioATP (2MeSATP), 2-chloroATP (2-ClATP), adenosine 5'-O-2-thiodiphosphate, (ADPbetaS), ATP and ADP, but not UTP, potentiated noradrenaline-induced N'-acetyl-5-HT production in a concentration-dependent manner. 2MeSATP neither induced the production of adenosine 3':5'-cyclic monophosphate (cyclic AMP), nor inhibited its formation when the glands were stimulated by forskolin. The phospholipase C inhibitor 1-[6-[[(17beta)-3-Methoxyestra-1,3,5(10)-trien-17-yl]amino]hexyl]-1H-pyrrole-2,5-dione (U73122), but not the inactive analogue, 1-[6-[[(17beta)-3-Methoxyestra-1,3,5(10)-trien-17-yl]amino]hexyl]-2,5-pyrrolidinedione (U73343), blocked the 2MeSATP effect. The P2 receptor antagonist pyridoxalphosphate-6-azophenyl-2',4'-dissulphonic acid (PPADS), which inhibits phospholipase C-coupled P2Y(1) receptors, blocked the 2MeSATP effect. In conclusion, our data strongly suggest that the P2-like receptor that is present in rat pinealocytes and which is responsible for the potentiation of noradrenaline-induced N'-acetyl-5-HT production is a P2Y(1)-like receptor, coupled to a G protein which stimulates phospholipase C.

Adenosine Diphosphate↗
PubMed2001

Pinealectomy-associated decrease in ribosomal gene activity in rats.

A decrease in ribosomal gene activity is an essential feature of the aging process as it was observed in Alzheimer's disease, in elderly Down's patients and in elderly healthy people. It is well known that aging is also associated with a reduction in melatonin synthesis. We studied 24 male Wistar rats cytogenetically, by using Ag-stained NOR (6 three-month-old rats underwent pinealectomy and were studied after 20 days; 6 control rats of the same age; 6 three-month-old rats underwent pinealectomy and were studied after 8 months; 6 control rats of the same age). Our results indicate that the absence of the pineal gland leads to a decrease in NOR activity. Further studies are necessary to determine if pinealectomy in rats could provide an animal model for aging.

Aging↗
PubMed2001

Tertian and quartan fevers: temporal regulation in malarial infection.

The periodicity in the development of Plasmodium parasites in infected animals, including man, has been known for almost 100 years. In turn, this periodicity is a consequence of the synchronous maturation of the parasite during its intracellular development. The cyclic fever that characterizes malarial infections is the outward manifestation of the parasite development. Until recently, little was known about the mechanisms by which parasite synchronicity is established and maintained. This review surveys the recent literature bearing on two main questions. (1) What are the mechanisms involved in the process of parasite synchronicity? (2) Do the circadian rhythms of the host interfere with the parasite cycle?

Animals↗
PubMed2000

Purinergic and noradrenergic cotransmission in the rat pineal gland.

ATP is coreleased with noradrenaline in several noradrenergic synapses, and P2-like receptors were shown to be present in rat pineal glands. A new method of functional investigation was developed to assess the importance of both transmitters (noradrenaline and ATP) in eliciting the synthesis of melatonin and its precursor N'-acetyl-5-hydroxytryptamine (N'-acetyl-5-HT) through transmural electrical field stimulation of cultured pineal glands. Incubation with the beta-adrenoceptor antagonist propranolol (>10(-7) M) blocked almost completely the production of N'-acetyl-5-HT, whilst the P2 receptor antagonists pyridoxalphosphate-6 azophenyl-2',4'-disulfonic acid (PPADS, >3x10(-6) M) and suramin (>10(-6) M) blocked it partially. These findings indicate a physiologically relevant role for the purinergic cotransmission in this system.

Adenosine Triphosphate↗
PubMed2000

Characterization of beta-adrenoceptors responsible for venom production in the venom gland of the snake Bothrops jararaca.

We have shown that the stimulation of beta-adrenoceptors is an important step in venom production in the Bothrops jararaca venom gland. In the present study, the pharmacological profile of the beta-adrenoceptor present in Bothrops jararaca venom gland was characterized by radioligand binding assay and by the ability of isoprenaline to promote accumulation of cyclic AMP in dispersed secretory cells. In both cases, the venom glands were obtained from non-extracted snakes (quiescent stage) or from snakes which venom was extracted 4 days before sacrifice (venom production stimulated stage). [125I]-iodocyanopindolol ([125I]-ICYP) bound to extracted gland membranes in a concentration-dependent and saturable manner, but with low affinity. Propranolol, beta1- or beta2-selective adrenoceptors ligands displaced the [125I]-ICYP binding with low affinity, while selective beta3-adrenoceptor ligands did not displace the [125I]-ICYP binding. The displacement of [125I]-ICYP by propranolol was similar in non-extracted and extracted glands, showing the presence of beta-adrenoceptors in both stages. In dispersed secretory cells of non-extracted glands, isoprenaline (1 microM) increased the cyclic AMP production and propranolol (10 microM) was able to block this effect. On the other hand, in extracted glands, isoprenaline had no effect. The results suggest that the beta-adrenoceptors present in the Bothrops jararaca venom glands are different from those (beta1, beta2 or beta3) described in mammals, but are coupled to the Gs protein, like the known beta-adrenoceptor subtypes. Moreover, previous in vivo stimulation of venom production desensitizes the beta-adrenoceptors system and, although the receptors could be detected by binding studies, they are not coupled to the Gs protein, indicating that beta-adrenoceptors stimulation contributes to the initial steps of venom synthesis.

Adrenergic beta-1 Receptor Agonists↗
PubMed1999

Adenosine inhibits the renal plasma-membrane (Ca2+ + Mg2+)-ATPase through a pathway sensitive to cholera toxin and sphingosine.

Adenosine, a potent autacoid produced and released in kidneys, affects nearly all aspects of renal function, and an increase in cytosolic calcium has been implicated in adenosine effects. The aim of this work was to investigate whether adenosine modifies the calcium pump present in basolateral membranes of kidney proximal tubule cells. Adenosine exerts a biphasic influence on (Ca2+ + Mg2+)-ATPase activity. Inhibition occurs up to 0.1 microM and then gradually disappears as the adenosine concentration increases to 100 microM, an effect mimicked by the adenosine analog N6-cyclohexyladenosine, which preferentially binds to A1-type receptors. In contrast, the A2 receptor agonist 5', N-ethylcarboxamideadenosine is ineffective. The A1 receptor antagonist 8-cyclopentyl-1,3-dimethylxanthine blocks the inhibitory effect of 0.1 microM adenosine and stimulates (Ca2+ + Mg2+)-ATPase activity in the presence of 1 mM adenosine, a concentration high enough to occupy the low-affinity A2 receptors. Inhibition by adenosine increases as medium ATP is lowered to micromolar concentrations, is maintained in the presence of pertussis toxin, and is completely abolished with 0.1 microM cholera toxin or 1 microM sphingosine. The inhibitory effect of adenosine can be reproduced by guanosine 5'-[gamma-thio]triphosphate, inositol 1,4, 5-trisphosphate or the diacylglycerol analog 12-O-tetradecanoylphorbol 13-acetate. In conjunction with the selectivity for its analogs and for its receptor agonist, the concentration profile of adenosine effects indicates that both inhibitory (A1) and stimulatory (A2) receptors are involved. The results obtained with the toxins indicate that a pathway that is modulated by G-proteins, involves a phospholipase C and a protein kinase C, and is affected by local variations in adenosine concentrations participates in the regulation of the (Ca2+ + Mg2+)-ATPase resident in basolateral membranes of kidney proximal tubules.

Adenosine↗
PubMed1999

Melatonin modulation of presynaptic nicotinic acetylcholine receptors located on short noradrenergic neurons of the rat vas deferens: a pharmacological characterization.

Melatonin, the pineal hormone produced during the dark phase of the light-dark cycle, modulates neuronal acetylcholine receptors located presynaptically on nerve terminals of the rat vas deferens. Recently we showed the presence of high affinity nicotine-binding sites during the light phase, and low and high affinity binding sites during the dark phase. The appearance of the low affinity binding sites was due to the nocturnal melatonin surge and could be mimicked by exposure to melatonin in vitro. The aim of the present research was to identify the receptor subtypes responsible for the functional response during the light and the dark phase. The rank order of potency of agonists was dimethylphenylpiperazinium (DMPP) = cytisine > nicotine > carbachol and DMPP = nicotine = cytisine > carbachol, during the light and dark phase, respectively, due to an increase in apparent affinity for nicotine. Mecamylamine similarly blocked the DMPP response during the light and the dark phase, while the response to nicotine was more efficiently blocked during the light phase. In contrast, methyllycaconitine inhibited the nicotine-induced response only at 21:00 h. Since alpha7 nicotinic acetylcholine receptors (nAChRs) have low affinity for nicotine in binding assays, we suggest that a mixed population composed of alpha3ss4 - plus alpha7-bearing nAChR subtypes is present at night. This plasticity in receptor subtypes is probably driven by melatonin since nicotine-induced contraction in organs from animals sacrificed at 15:00 h and incubated with melatonin (100 pg/ml, 4 h) is not totally blocked by mecamylamine. Thus melatonin, by acting directly on the short adrenergic neurons that innervate the rat vas deferens, induces the appearance of the low affinity binding site, probably an alpha7 nAChR subtype.

Animals↗
PubMed1997

Age-related changes in the reactivity of postsynaptic adrenoceptors in the rat vas deferens: differences between the epididymal and prostatic portion.

1. Age-related changes in the reactivity of postsynaptic alpha-adrenoceptors of isolated portions (epididymal and prostatic) and in whole vas deferens have been studied using 4, 12 and 20 month-old rats. 2. The pD2 values for adrenaline-induced contractions were reduced in the epididymal portion and whole vas deferens of middle-aged and old animals, but not in the prostatic portion. No age related change to phenylephrine or clonidine sensitivity was observed. 3. pA2 values of prazosin and yohimbine were not changed by aging in any preparation. Phentolamine pA2 values were reduced in the epididymal portion and in the whole vas deferens when adrenaline, but not when phenylephrine concentration-response curves were displaced by the antagonist. The mean pA2 value of yohimbine (6.78) indicates that this antagonist blocks alpha(1)-adrenoceptors in the rat vas deferens. 4. The data presented here suggest that age-related decreases in the sensitivity to adrenaline and phentolamine (when measured by displacing adrenaline concentration-effect curves) in the whole vas deferens are probably due to a variation in the proportion of alpha(1)-adrenoceptor subtypes in the epididymal portion of the rat vas deferens.

Adrenergic alpha-Agonists↗
PubMed1997

Circadian rhythm in experimental granulomatous inflammation is modulated by melatonin.

Biological rhythms are detected in a variety of physiological and pathological conditions in man and animals, such as rheumatoid arthritis and asthma. Here we describe a circadian rhythm in experimental infectious and non-infectious granuloma. After 30 days of BCG (Bacillus Calmette-Guerin) or nystatin inoculation in the left hind foot of C57B1/6 mice, there is an oscillation with a period of approximately 24 hr in the variation of paw thickness, indicating a circadian rhythm. The acrophase occurred during the light phase, between 9:00 and 13:00 hr, while the nadir occurred in the dark phase, between 21:00 and 01:00 hr. The vascular permeability around the granulomatous lesions was higher at 12:00 hr than at 24:00 hr. This is in agreement with the observation that the thickness of a paw with granulomatous lesion is larger during the light phase. This rhythmic variation was eliminated by either pinealectomy or superior cervical ganglionectomy, which greatly reduce melatonin levels in the blood. Nocturnal replacement of melatonin in pinealectomized mice led to the re-establishment of the circadian rhythm. Thus, the rhythm of the granulomatous lesion is due to the rhythmic melatonin release by the pineal gland. This approach opens new questions regarding the modulation of chronic inflammation in inflammatory diseases that present rhythmic symptoms throughout the day.

Animals↗
PubMed1997

Control of venom production and secretion by sympathetic outflow in the snake Bothrops jararaca.

Many studies have examined the morphological and biochemical changes in the secretory epithelium of snake venom glands after a bite or milking. However, the mechanisms of venom production and secretion are not yet well understood. The present study was undertaken to evaluate the role of the sympathetic nervous system in the control of venom production and secretion. Venom glands were obtained from Bothrops jararaca (Viperidae) snakes, either unmilked previously or milked 4, 7 or 15 days before they were killed. Levels of tyrosine-hydroxylase-like immunoreactivity were higher in venom glands collected 4 days after milking, coinciding with the maximal synthetic activity of the secretory cells. The only catecholamine detected by high-performance liquid chromatography was noradrenaline, indicating the presence of noradrenergic fibres in these glands. In reserpine-treated milked snakes, no venom could be collected, and electron microscopic analysis showed narrow rough endoplasmic reticulum cisternae, instead of wide cisternae, and less well-developed Golgi apparatus compared with milked untreated snakes, indicating impairment of protein synthesis and secretion. The administration of isoprenaline or phenylephrine (beta- and alpha-adrenoceptor agonists, respectively) to reserpine-treated milked snakes promoted the widening of the rough endoplasmic reticulum and restored venom production, but only phenylephrine restored the development of the Golgi apparatus and the formation of many secretory vesicles. These results provide the first evidence that the sympathetic nervous system plays an important role in venom production and secretion in the venom glands of Bothrops jararaca. Understanding the importance of noradrenergic stimulation in venom production may provide new insights for research into the treatment of snakebites.

Animals↗
PubMed1996

Are imidazoline receptors involved in sympathetic neurotransmission in rat vas deferens?

1. An involvement of imidazoline receptors in the modulation of neurotransmitter release was investigated in the prostatic portion of the rat vas deferens stimulated transmurally at 0.2 Hz or by single pulses. 2. Idaxozan and yohimbine induced a concentration-dependent potentiation of the contractile response to 0.2-Hz transmural stimulation in the epididymal and prostatic portion of the vas. 3. After reserpine treatment, idazoxan, but not yohimbine, still potentiated the contractile response, suggesting a possible involvement of imidazoline receptors. 4. Clonidine and rilmenidine, agonists with different affinities to alpha 2-adrenoceptors and imidazoline receptors, inhibited with the same potency the contractile responses to a single pulse transmural stimulation. 5. Yohimbine (a selective alpha 2-adrenoceptor antagonist) antagonized the inhibitory concentration effect curve to rilmenidine in a competitive manner. pA2 values for idaxozan (an antagonist to alpha 2-adrenoceptors and imidazoline receptors) were not different when noradrenaline or rilmenidine were used as agonists. Phenoxybenzamine blocked the effect of both agonists. 6. Thus, the potency relationship of agonists, as well as the effect of the antagonists, did not favor the hypothesis that imidazoline receptors are involved in the idazoxan-potentiating effect in the rat vas deferens.

Adrenergic alpha-2 Receptor Agonists↗
PubMed1996

Melatonin modulation of presynaptic nicotinic acetylcholine receptors in the rat vas deferens.

Melatonin, the pineal hormone produced during the dark phase of the light-dark cycle, modulates neuronal acetylcholine receptors located presynaptically on the sympathetic nerve terminals of the rat vas deferens. The pD2 values for nicotine were significantly higher at night (4.20 +/- 0.01) than in the afternoon (3.80 +/- 0.07). Exogenous melatonin shifted the concentration-response curves for nicotine to the left, mimicking the effect of darkness. Melatonin modifies both the displacement and the saturation curves of [3H](-)nicotine binding. In membranes from animals killed at 15:00 h, the binding of [3H](-)nicotine (5-6 nM) was first potentiated and then inhibited by sequential concentrations of (-)nicotine. Higher concentrations of [3H](-)nicotine (50-60 nM) were displaced by all concentrations of nonlabeled ligand. However, when membranes from tissues exposed to melatonin (exogenous or endogenous) were tested, the lower [3H](-)nicotine concentration was displaced progressively by increasing concentrations of nonlabeled ligand. Equilibrium binding studies show a single class of high-affinity nicotinic binding sites with an apparent Kd value of 16 nM and an average maximal number of binding sites of 66 fmol mg-1 protein when animals were killed at the afternoon. Melatonin, although it did not change the properties of high-affinity binding sites, induced the appearance of a second population of lower apparent affinity (Kd = 36.7 nM; Bmax = 185.4 fmol/mg). Melatonin does not modify the functional response and the displacement of [3H](-)nicotine by dimethylphenylpiperazinium. The data suggest that nicotinic neuronal acetylcholine receptors stimulated by dimethylphenylpiperazinium do not change between the light and dark phases. Rather, the higher sensitivity to nicotine in prostatic portions incubated with exogenous melatonin, and in organs from animals killed at night, after the rise of endogenous melatonin, is probably due to the appearance of low-affinity neuronal nicotinic ACh binding sites.

Acetylcholine↗
PubMed1994

Modulation of sympathetic neurotransmission by melatonin.

Melatonin is of considerable interest for its regulatory influence on a variety of physiological processes including biological rhythms and neuroendocrine functions. We showed that melatonin potentiates sympathetic neurotransmission in the prostatic portion of the rat vas deferens, by increasing contractions in response to noradrenaline and ATP released by acetylcholine stimulation of presynaptic nicotinic receptors. Melatonin in vitro (100 pg/ml; for 4 h) increased the maximal acetylcholine-induced contraction only when the hypogastric ganglion was present, and this effect was blocked by cycloheximide (100 micrograms/ml). Melatonin also modulated the sympathetic trophic influence on smooth muscle, since it reduced [35S]methionine incorporation into the vas deferens in the hypogastric ganglion-vas deferens preparation. Thus, it is suggested that the regulation of protein synthesis might be one of the mechanisms whereby melatonin modulates endogenous rhythms and synchronizes them to the environmental light cycle.

Acetylcholine↗
PubMed1994

Presence of P2-purinoceptors in the rat pineal gland.

1. The effects of noradrenaline, ATP, adenylyl-imidodiphosphate (AMP-PNP), adenosine, alpha,beta-methylene-ATP and the P2-purinoceptor antagonist, suramin on N'-acetyl-5-hydroxytryptamine production were studied in cultured denervated rat pineal glands. 2. Noradrenaline (3 nM-1 microM) increased N'-acetyl-5-hydroxytryptamine production as measured both in the gland and the culture medium. 3. In noradrenaline (10 nM)-stimulated pineal glands, ATP (0.03 nM-1 mM) or AMP-PNP (0.1 microM-1 mM) increased N'-acetyl-5-hydroxytryptamine production in a concentration-dependent manner. 4. Alpha,beta-Methylene-ATP at the concentration of 0.1 mM, but not 3 microM, attenuated the enhancement by ATP (0.1 mM) of noradrenaline (10 nM)-induced N'-acetyl-5-hydroxytryptamine production. 5. Suramin (0.1 mM) blocked the potentiating effect of ATP (0.1 mM), but not the potentiating effect of adenosine (0.1 mM) in glands incubated with noradrenaline (10 nM). 6. These findings suggest that the rat pineal gland possesses P2-purinoceptors which when stimulated potentiate the effect of noradrenaline but do not, by themselves, induce an increase in N'-acetyl-5-hydroxytryptamine production.

Adenosine↗