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Biomedical subjects

U Ackermann

Publications and source records attributed to U Ackermann.

At least 37 records · Page 2Linked to original sources

Cloning and expression analysis of Aedes aegypti opsin: adaptation of an in situ hybridization protocol for mosquitoes.

Opsin is a G protein coupled photoreceptor that activates a signal transduction cascade in the ommatidia. Its primary and secondary structure is conserved both in insects and vertebrates as exemplified by the Drosophila opsins. Through serendiplious cloning of a PCR fragment, we have identified an opsin cDNA. The latter was used to clone full length cDNAs from a mosquito head library. The main purpose of cloning was to have a positive control probe to establish an in situ hybridization protocol for less abundant probes. Opsin-mRNA is localized specifically to the visual receptor cells in the ommatida. No other cells in the brain or the remainder of the body are positive. This is confirmed by Northern blot analysis. The sequence of the receptor, of which we have found two different transcripts, confirms its typical topology, including the seven transmembrane spanning regions and the intracellular carboxy terminus that has potential phosphorylation sites. Our in situ hybridization protocol combines several procedures: the most important points are: (a) the immediate processing of sections after cutting, and (b) the sections are never allowed to dry out once the procedure was started. Our protocol has a much higher sensitivity, using approximately 50 x lower concentrations of probe compared to published protocols. In addition to the detection of opsin-mRNA, it has been successfully applied to the detection of the low abundant insulin receptor homologue. Furthermore, Aedes aegypti probes were visualizing a similar tissue specificity when applied to the malaria mosquito Anopheles albimanus.

Aedes↗

Potassium depletion and salt-sensitive hypertension in Dahl rats: effect on calcium, magnesium, and phosphate excretions.

Weanling male inbred Dahl rats (Jr salt-sensitive (S) and salt-resistant (R) strains) were placed on high (4%, HK) and low (0.2%, LK) potassium diets for 4 weeks. Both diets contained 8% sodium chloride, 2.5% calcium, 0.8% magnesium, and 2.0% phosphorous. Balance studies were carried out during the final week on the diets. Mean arterial blood pressure was determined, and dietary intake and urinary output of water, sodium, chloride, potassium, calcium, magnesium, and phosphate were monitored daily during this period. The data show that blood pressures of S rats were significantly higher than those of R rats on both HK and LK diets; however, reduced dietary potassium was associated with increased blood pressure in both strains. Urinary excretions of calcium and magnesium were higher, and urinary phosphate excretion was lower, in S compared to R rats. Decreased potassium intake was associated with increased excretion of calcium, magnesium and phosphate in both strains. The changes in calcium and magnesium excretion were significantly correlated to blood pressure across strains and diets. We conclude that the effects of a high salt diet on increasing blood pressure can be potentiated by lack of potassium, even in previously salt-resistant rats. Increased blood pressure is associated with increased divalent cation excretion. It is not yet known whether this is a cause-and-effect relationship.

Animals↗

The interaction between atrial natriuretic peptide and cardiac parasympathetic function.

We have demonstrated previously that atrial natriuretic peptide (ANP) inhibits hypotension-induced reflex tachycardia via a parasympathetic mechanism. The present study further defines that parasympathetic mechanism. We tested the hypothesis that ANP, during vagus nerve stimulation, acts as a physiological antagonist to interfere with alpha 1-adrenoceptor modulation of efferent cardiac vagal action. Sprague Dawley rats were divided into five groups, each group receiving a different infusion. Infusates included one of vehicle (Ringer's solution; RS), an alpha 1-adrenoceptor agonist (phenylephrine; PE), a combination of agonist and either a known alpha 1-adrenoceptor antagonist (prazosin; PE+PRZ) or the putative physiologic antagonist, ANP (PE+ANP). The fifth group received all three drugs, PE+PRZ+ANP. Under Inactin anesthesia (100 mg/kg i.p.), efferent autonomic input to the heart was surgically interrupted. Animals were also adrenalectomized to limit the effects of circulating catecholamines. We then monitored each group for the change in heart rate (delta HR) in response to efferent vagus nerve stimulation at various frequencies (2 Hz, 5 Hz, 10 Hz). Infusion of PE significantly (P < 0.01 by ANOVA) attenuated the magnitude of delta HR when compared to the RS group. This attenuation of vagally-induced bradycardia was eliminated by the addition of the alpha 1-adrenoceptor antagonist, prazosin (PE+PRZ group). The PE+ANP group responded with results similar to those of the PE+PRZ group. There was no difference between delta HR responses of the PE+PRZ+ANP group and the PE+PRZ group.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenergic alpha-Antagonists↗

ANP has no postsynaptic effect on autonomic regulation of cardiac pacemaker rate in the rat.

We studied whether atrial natriuretic peptide (ANP) influences sinoatrial node pacemaker activity or whether it modifies the response to activation of postsynaptic autonomic receptors. Male Sprague-Dawley rats were anesthetized with pentobarbital sodium (45 mg/kg). Their hearts were removed quickly and placed in physiological salt solution. The atria were isolated; the right intra-atrial chamber was exposed to allow intracellular recording from sinoatrial node pacemaker cells. The tissue was placed in a temperature-regulated recording chamber and superfused with warmed oxygenated physiological salt solution. With use of standard microelectrode recording techniques, action potentials were recorded from spontaneously depolarizing cells in the presence of muscarine (62.5-500 nM) or norepinephrine (0.1 and 1.0 microM). Muscarine reduced the frequency of action potentials dose dependently, whereas norepinephrine increased their frequency. The addition of ANP (0.1-100 nM) to the superfusion had no effect on the frequency of action potentials during the superfusion of physiological salt solution or in the presence of either muscarine or norepinephrine. We conclude that ANP does not act on cardiac pacemaker cells to modulate the effect of neurotransmitters.

Action Potentials↗

Beta 1- and beta 3-subunits can associate with presynthesized alpha-subunits of Xenopus oocyte Na,K-ATPase.

Oligomerization of newly synthesized alpha- and beta-subunits is a prerequisite for the structural and functional maturation of Na,K-ATPase. In this study, we have tested the competence of presynthesized alpha- and beta-subunits to assemble into functional enzyme complexes. Antisense oligonucleotides complementary to alpha-mRNA were used to inhibit alpha-subunit synthesis in Xenopus oocytes leaving a presynthesized trypsin-sensitive alpha-subunit pool. beta-Subunits expressed in these oocytes from injected cRNA assembled with the preexisting alpha-subunits, rendered them trypsin-resistant, and permitted the expression of more ouabain binding sites at the plasma membrane. Similarly, presynthesized beta 1- or beta 3-subunits produced in Xenopus oocytes by injection of beta-cRNA and later of specific antisense oligonucleotides were stabilized and transported out of the endoplasmic reticulum when alpha-cRNA was injected into oocytes. These data indicate that alpha- and beta-subunits can insert into endoplasmic reticulum membranes independent of each other in an assembly-competent form and retain their ability for oligomerization after synthesis.

Animals↗

Atrial natriuretic peptide inhibits compensatory responses when cardiac performance is depressed.

We tested the hypothesis that the atrial natriuretic peptide (ANP) mediated decrease in baroreceptor sensitivity that is seen in normal rats is more pronounced in a state of depressed cardiac performance. Holtzman rats (n = 15) were injected with Adriamycin (1 mg/kg i.p. 3 times/week for 8-10 weeks). Control rats (n = 17) were injected with 0.9% saline. Experiments were done in conscious animals that had been catheterized for i.v. infusions and for measurement of arterial blood pressure (ABP) and heart rate (HR). ANP (250 ng.kg-1.min-1) or saline vehicle was infused i.v. Graded periodic bolus injections of phenylephrine or sodium nitroprusside were given to assess baroreceptor sensitivity (beats.min-1.mmHg-1) up to 60 mmHg (1 mmHg = 133.3 Pa) above and below resting ABP. The following day the experiment was repeated with the ANP-vehicle regimen reversed. Finally, the rats were anesthetized and the rate of left ventricular pressure increase (dP/dt) was measured. Data evaluation included calculation of least squares linear regression slopes of peak delta HR vs. peak delta ABP, applying corrections for experimental errors in both the dependent and independent variables. Adriamycin rats (A) did not differ significantly from control rats (C) with respect to either initial ABP (A = 105 +/- 5; C = 100 +/- 3; mean mmHg +/- SEM) or initial HR (335 +/- 9 vs. 312 +/- 13 beats.min-1). However, their indices of cardiac performance were significantly depressed.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Modulation of Na,K-ATPase expression during early development of Xenopus laevis.

In amphibian and mammalian systems, regulation of Na+ transport via the Na,K-ATPase plays an important role in distinct developmental processes such as blastocoele formation and neurulation. In this study, we have followed the Na,K-ATPase activity, the biosynthesis, and the cellular accumulation of catalytic alpha-subunits after fertilization of Xenopus laevis eggs up to neurula formation. Our data show that Na,K-ATPase activity increases significantly between stages 4 and 6 and again between stages 13 and 24. The four-fold rise in Na,K-ATPase activity during blastocoele formation is not mediated by an increased cellular pool of alpha-subunits. On the other hand, a five-fold increase of the biosynthesis rate around midblastula precedes a progressive accumulation up to neurula stage mainly of alpha 1-subunits and to a lesser extent of a second alpha-immunoreactive species. In contrast, newly synthesized glycoproteinic beta 1-subunits of Na,K-ATPase cannot be detected up to late neurula. These data indicate that (1) upregulation of Na,K-ATPase activity during blastocoele and neurula formation are mediated by different regulation mechanisms and (2) alpha- and possibly beta-isoforms are expressed in a developmentally regulated fashion during early Xenopus development.

Animals↗

[Correlation between respiratory tract symptoms in young children and NO2 concentration of outside air].

The aim of a one year study was to ascertain whether air pollution measured as NO2-concentration has an measurable influence on the health of infants in Switzerland. Measurements of NO2-concentration were carried out by means of passive collectors in ambient air, living room and at the child itself. The results of 1225 children printed to a significant coherence between respiratory symptoms per day and child and the NO2 load of ambient air as an guide pollutant. Even in consideration of other childish, familiar and environmental factors the coherence remains. Although no causal relation thereby proved this factor supports the observation of parents and physicians that children more often fall ill of respiratory diseases in polluted air being in accordance with similar investigations abroad.

Air Pollutants↗

Mutual dependence of Na,K-ATPase alpha- and beta-subunits for correct posttranslational processing and intracellular transport.

In this study, we have followed the fate of newly synthesized alpha- and beta-subunits of Na,K-ATPase in Xenopus oocytes injected with alpha and/or beta cRNA to examine whether assembly of the two subunits is needed for a correct folding and/or for intracellular transport of Na,K-ATPase. Our data indicate that (1) assembly of alpha- and beta-subunits occurs at the level of the ER, (2) beta-subunits are needed for the newly synthesized alpha-subunit to adopt a stable configuration and (3) alpha- and beta-subunits mutually depend on each other to be transported out of the ER.

Animals↗

Significant association between outdoor NO2 and respiratory symptoms in preschool children.

A study of 1225 preschool children was conducted in four regions of Switzerland with different levels of air pollution to investigate the relationship between air pollution and respiratory symptoms. Daily symptoms were recorded by parents on a diary form and air pollution exposure assessed by personal NO2 samplers. Each family participated for 6 weeks and personal samplers were changed every week. The frequency of respiratory symptoms per child and day was found to increase with increasing levels of NO2 measured outdoors. This relationship remained significant in a multiple regression model in which the factors smoking, origin, indoor air pollution, age and sex, season, and parents appreciation of air pollution at the living site were taken into account.

Air Pollutants↗

Influence of atrial natriuretic factor on autonomic control of heart rate.

Atrial natriuretic factor (ANF) may prevent reflex tachycardia. To determine whether this action occurs in the central or peripheral nervous system, Sprague-Dawley rats, anesthetized with Inactin (100 mg/kg ip), underwent surgical cardiac denervation and subsequent vagus nerve and stellate ganglion stimulation. We compared the change in heart rate (delta HR) in response to high- and low-frequency stimulation in a group receiving a vehicle infusion with a group receiving an ANF infusion (0.28 microgram.kg-1.min-1). We found that ANF had no effect on delta HR in the absence of all stimulation or during sympathetic stimulation at any frequency. There was no significant interaction between ANF enhancement of parasympathetic activity and the level of sympathetic stimulation. ANF acted peripherally to significantly (P less than 0.05) increase the magnitude of delta HR in response to parasympathetic stimulation at high (5 Hz) and low (2 Hz) frequencies. We speculate that a common step in mechanism of action of ANF and acetylcholine may be responsible for the enhancement of cardiac parasympathetic effects by ANF.

Animals↗

[Correlation of respiratory tract symptoms in young children and NO2 concentrations of the outside air].

In four regions of Switzerland (2 towns, 1 municipal agglomerate, 1 rural area) the relationship between the degree of air pollution measured in terms of NO2 and the incidence of airway symptomatology was investigated in 1,225 young children. The airway symptoms were recorded by the parents in a diary; the NO2 loading was measured with the aid of personal collecting tubes at the place of residence of the child. A significant relationship was found to exist between the mean incidence of airway symptomatology per child and day, and the individually measured NO2 concentration in the outside air at the child's place of residence. This relationship remained significant even when, in a multiple regression analysis, account was also taken of other major factors, such as smoking, nationality, individual susceptibility to airway diseases, the season of the year, and the subjective assessment of the air pollution at the place of residence, were also taken into account.

Air Pollutants↗

Angiotensin restores atrial natriuretic factor-induced decrease of baroreceptor sensitivity in normotensive rats, but not in spontaneously hypertensive rats.

The effect of atrial natriuretic factor (ANF) on baroreflex sensitivity was determined in unanesthetized normotensive (Wistar-Kyoto, WKY) or spontaneously hypertensive rats (SHR) during acute hypertensive stimuli (phenylephrine) or hypotensive stimuli (sodium nitroprusside). The i.v. dose of rat ANF [( Ser99,Tyr126]ANF) was 50 ng/min per rat, sufficient to decrease mean arterial blood pressure (ABP) by about 6 mmHg (1 mmHg = 133.3 Pa) in WKY. SHR showed no change in ABP with this ANF dose. During a control infusion of physiological saline, the mean heart rate (HR) response to increases in ABP was -1.30 +/- 0.27 beats/min (bpm)/mmHg in WKY and -0.37 +/- 0.22 in SHR (p less than 0.05). These values were not affected significantly by ANF. However, ANF blunted chronotropic responses to ABP decreases. The control values of the delta HR/delta ABP slope in WKY and SHR were -2.34 +/- 0.57 and -2.01 +/- 0.37 bpm/mmHg, respectively. In the presence of ANF, the slope changed to -0.36 +/- 0.43 (i.e., bradycardia in response to hypotension) in WKY and to +0.20 +/- 0.21 in SHR (p less than 0.005 for the difference from control for both). This ANF-induced loss of baroreflex sensitivity was reversed in WKY by the addition of angiotensin I (sufficient to increase ABP by 5 mmHg in control rats). Angiotensin did not restore baroreflex sensitivity in ANF-infused SHR, and ANF had no effect on the ABP increase caused by angiotensin in either group. The data suggest that ANF does not act on baroreceptor structures directly, but inhibits mechanisms involved in efferent sympathetic activation. Parasympathetic responses do not appear to be compromised.

Angiotensin I↗

Atrial natriuretic factor alters autonomic interactions in the control of heart rate in conscious rats.

Regulation of heart rate was studied in rats receiving either i.v. saline at 64 microL/min or synthetic 28-residue rat atrial natriuretic peptide (ANF) at a dose sufficient to decrease mean arterial blood pressure by 10%. Autonomic influences were deduced from steady-state heart rate responses of each group to propranolol, atropine, or propranolol and atropine combined. A multiplicative model of heart rate control was used to derive quantitatively from the data the modulation of intrinsic heart rate by sympathetic and parasympathetic mechanisms. Animals receiving ANF showed a lower heart rate than control animals. This relative bradycardia was abolished by atropine. Blocking of sympathetic effects with propranolol had no effect on basal heart rate in either group, and atropinization led to significant increases in heart rate in both groups of rats. Mathematical analysis of the results showed that the bradycardia produced by ANF was due predominantly to a reduced intrinsic heart rate and to enhanced vagal inhibition of postganglionic sympathetic activity. Parasympathetic contribution to heart rate in the absence of sympathetic activity was negligible in control rats and small during ANF. We conclude that the major influences of ANF on heart rate control are a decrease of intrinsic heart rate and enhanced parasympathetic inhibition of postganglionic presynaptic sympathetic activity.

Animals↗

Cardiovascular effects of atrial natriuretic extract in the whole animal.

Atrial tissue extract (AE) and ventricular tissue extract cause identical decreases in total peripheral resistance when they are injected i.v. into anesthetized rats. However, only AE causes significant hypotension because of cardiac inhibition. This involves both bradycardia and failure of stroke volume to increase appropriately. The observations cannot be explained by direct action of AE on myocytes, but are more likely to be the result of interactions with cardiovascular reflex mechanisms. Excitation of chemosensitive cardiac receptors with vagal afferents appears to be an important afferent mechanism. The efferent limb for the negative chronotropic response resides partly in the vagus nerves and partly in cardiac sympathetic nerves. The negative inotropic response of AE was not altered by vagotomy, spinal section, atropine, or propranolol. These results suggest that atrial peptides may cause the release of a negatively inotropic substance from a site that is not yet identified.

Afferent Pathways↗

Renal venous pressure and volume natriuresis in the rat.

Anesthetized rats were surgically prepared so that left renal venous pressure could be maintained at its normal level (near 1 mmHg) (1 mmHg = 133.322 Pa) while right renal venous pressure was free to change in response to acute isoncotic blood volume expansion (n = 10). The purpose of the study was to test whether the renal venous pressure changes normally accompanying a 33% blood volume expansion have an effect on inulin clearance (CIn) and on the rates of excretion of water (V) or sodium (UNaV) by the kidneys. In 10 control rats blood volume was not expanded. The results showed that the volume expansion used resulted in a peak pressure of 5.3 +/- 0.5 (SEM) mmHg in the right renal vein. Both kidneys showed significant increases in CIn, V, and UNaV over their respective preexpansion values. However, there were no significant differences between these parameters measured in the right or in the left kidney even though the differences between the two renal venous pressures were highly significant during the infusion and for the next 20 min. In a further six rats, a caval snare was placed superior to the right renal vein so that both renal venous pressures could be held at normal during blood volume expansion. The ensuing diuresis and natriuresis were not different from those observed in the preceding group. It is concluded that changes in renal venous pressure, of the magnitude associated with volume expansion, have no significant effect on renal excretion rates.

Animals↗