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R Jahns

Publications and source records attributed to R Jahns.

18 recordsLinked to original sources
PubMed1997

Probing human beta1- and beta2 -adrenoceptors with domain-specific fusion protein antibodies.

In order to generate antibodies suitable for immunological studies on beta-adrenoceptors constitutively expressed at low levels in cells or tissues we have produced fusion proteins of the amino- and carboxy-terminus, and the second extracellular loop of the human beta1- or beta2-adrenoceptors with bacterial glutathione-S-transferase in E. coli. Rabbit antibodies raised against these fusion proteins strongly reacted with intact human beta1- or beta2-adrenoceptors in a subtype- and domain-specific manner. Antibodies directed against the second extracellular loop of the beta1-adrenoceptor reacted stronger with non-denatured receptors and decreased the affinity of the 3H-labelled antagonist (-)-4-(3-t-butylamino-2-hydroxypropoxy)-[5,7-3H]benzimidazol-2-one ([3H]CGP 12 177), indicating a specific interaction with the native receptor. In contrast, antibodies directed against carboxy- and amino-terminal receptor domains reacted strongly both with denatured and non-denatured receptors but did not interfere with binding of [3H]CGP 12 177. Affinity purified antibodies were used for detecting the beta1- or the beta2-adrenoceptor subtype heterologously produced in Sf9 cells by enzyme-linked immunosorbent assay, Western blotting, immunoprecipitation, and indirect immunofluorescence microscopy. Moreover, we could demonstrate that avidity, titers, and specificity of these antibodies were high enough for studying beta-adrenoceptors constitutively expressed in human A431 cells, where we observed a patched membrane distribution of the receptors.

Animals↗
PubMed1997

Regulation of Na+/H+ antiporter in trout red blood cells.

The trout red blood cell Na+/H+ antiporter (beta NHE) plays two interesting properties: it is the only NHE own to be activated by cyclic AMP, and the activation process is followed by a desensitisation of the transport system itself. Cloning and expression of beta NHE have provided inificant information about Na+/H+ activation, in particular that activation by cyclic AMP is directly dependent upon the presence of two protein kinase A consensus sites in the cytoplasmic tail of the antiporter. Expression of beta NHE in fibroblasts demonstrates that the protein kinase A (PKA) and protein kinase C (PKC) activation pathways are independent and do not converge a common kinase. Moreover, the hydrophilic C-terminal fragment is essential to the mediation of the various hormonal responses. NHE1 (the human ubiquitous isoform) is not activated by cyclic AMP, but a "NHE1 transmembrane domain/beta NHE cytoplasmic domain' chimera is fully activated by cyclic AMP. In red cells, activation of beta NHE is the result of phosphorylation by PKA of at least two independent sites. Desensitisation, inhibited by the phosphatase inhibitor okadaic acid, may consist of the dephosphorylation of one of these two sites. Furthermore, Calyculin A (CIA), another specific protein phosphatase inhibitor, induces in unstimulated cells a Na+/H+ exchange activity whose exchange properties are very different from those of the adrenergically stimulated antiporter. It is suggested that CIA may be able to revive "sequestered' antiporters. We propose that the molecular events underlying beta NHE desensitisation could be similar to those involved in rhodopsin desensitisation. Antibodies were generated against trout red cell arrestin in order to analyse the binding of arrestin to the activated exchanger. Recombinant trout arrestin was produced in a protease-deficient strain of Escherichia coli and its functionality tested in a reconstituted rhodopsin assay.

Amino Acid Sequence↗
PubMed1996

Probing human beta 1- and beta 2-adrenoceptors with domain-specific fusion protein antibodies.

In order to generate antibodies suitable for immunological studies on beta-adrenoceptors constitutively expressed at low levels in cells or tissues we have produced fusion proteins of the amino- and carboxy-terminus, and the second extracellular loop of the human beta 1- or beta 2-adrenoceptors with bacterial glutathione-S-transferase in E. coli. Rabbit antibodies raised against these fusion proteins strongly reacted with intact human beta 1- or beta 2-adrenoceptors in a subtype- and domain-specific manner. Antibodies directed against the second extracellular loop of the beta 1-adrenoceptor reacted stronger with non-denatured receptors and decreased the affinity of the 3H-labelled antagonist (-)-4-(3-t-butylamino-2-hydroxypropoxy)-[5,7-3H]benzimidazol-2-one ([3H]CGP 12 177), indicating a specific interaction with the native receptor. In contrast, antibodies directed against carboxy- and amino-terminal receptor domains reacted strongly both with denatured and non-denatured receptors but did not interfere with binding of [3H]CGP 12 177. Affinity purified antibodies were used for detecting the beta 1- or the beta 2-adrenoceptor subtype heterologously produced in Sf9 cells by enzyme-linked immunosorbent assay, Western blotting, immunoprecipitation, and indirect immunofluorescence microscopy. Moreover, we could demonstrate that avidity, titers, and specificity of these antibodies were high enough for studying beta-adrenoceptors constitutively expressed in human A431 cells, where we observed a patched membrane distribution of the receptors.

Animals↗
PubMed1996

Trout red blood cell arrestin (TRCarr), a novel member of the arrestin family: cloning, immunoprecipitation and expression of recombinant TRCarr.

Arrestins are cytosolic proteins involved in the desensitization of G-protein-coupled receptors. We report the cloning of trout red blood cell arrestin which shows 76, 82 and 52% identity with bovine beta-arrestin1, beta-arrestin2 and retinal arrestin respectively. Antibodies were generated against the C-terminus of trout red blood cell arrestin. These antibodies detected arrestin in erythrocyte cytosol and were able to precipitate the native protein. The Na+/H+ antiporter of trout red blood cell is activated by beta-adrenergic stimulation and is then desensitized whereas the transmembrane signalling pathway is not. To investigate the subcellular distribution of arrestin on beta-adrenergic activation and desensitization of the antiporter, precipitation experiments were carried out on trout erythrocytes. A desensitization-dependent shift in cytosolic arrestin to the membranes could not be detected using the immunoprecipitation technique but we cannot exclude the possibility that a small number of cytosolic arrestins might be involved in the regulation of membrane proteins in trout erythrocyte. Recombinant trout arrestin was produced in a protease-deficient Escherichia coli strain and its functionality was tested in a reconstituted rhodopsin assay. The recombinant protein provides a suitable tool for investigating the target for arrestin in trout red blood cell, which still remains to be identified.

Amino Acid Sequence↗
PubMed1994

[Analysis of the pressure-surface relation of the left ventricle with automatic echocardiographic contour detection].

UNLABELLED: Echocardiographic automatic border detection is a new on-line technique determinating the interface between blood and myocardial tissue thus having the potential to calculate cyclic changes in left ventricular cavity area in real time. It was the main purpose of the current study to evaluate left ventricular pressure-area relationship after administration of nitrates. In 12 patients with normal left ventricular function pressure-area relation was studied after a Swan-Ganz thermodilution catheter was placed in the wedge position and a high fidelity pig tail catheter was placed in the left ventricle. Left ventricular pressure and cyclic changes of cavity area were simultaneously analysed and displayed together as waveforms on the echo screen using a computer interfaced with the echo machine. All measurements were done before and five minutes after administration of 0.8 mg nitroglycerin. Mean systolic and diastolic blood pressure decreased significantly from 145/12 mmHg to 127/8 mmHg (p < 0.05). Mean systolic area decreased slightly from 10 cm2 to 9 cm2 (n.s.) whereas mean enddiastolic area decreased significantly from 18 cm2 to 15 cm2 (p < 0.05). Accordingly there was a downward and leftward shift of the diastolic pressure-area relationship following administration of nitroglycerin. CONCLUSION: Echocardiographic automated border recognition seems to be a promising new on-line method in the detection of left ventricular cavity area changes underlining its potential usefulness in the evaluation of left ventricular performance.

Adult↗
PubMed1975

Types of neuronal responses in the rat thalamus to peripheral temperature changes.

Recordings were made of spike-trains from 163 neurons of the rostral part of the ventrobasal thalamus complex of the rat while the temperature of the scrotal skin was altered. The following results were obtained: 55 neurons were non-thermosensitive, 7 neurons cold-sensitive and 101 neurons warm-sensitive. In the case of the warm-sensitive cells a definite discrimination was possible: 61.4% of the neurons altered their firing behavior during peripheral cooling, changing from relatively even spike frequency to burst firing. This change could be induced repeatedly by altering the temperature. 38.6% of the neurons, on the other hand, reacted to cooling by slowing down their frequency. By way of statistical methods the firing patterns of the two response-types were analyzed more precisely and the differences in response to temperature stimuli more exactly defined. Likewise, the spatial distribution of the two response-types of warm-sensitive cells exhibited differences; whereas the cells devoid of burst activity occured rather uniformly in the region of the thalamus studied, the cells with bursting activity were confined more to the mediocaudal region. These findings are discussed with regard to the phenomenon of peripheral bursts and to the projection of thermoafferent pathways onto the ventrobasal thalamus complex. The functional interpretation of the various cell reactions and their role in the central processing of thermoafferent signals still remains unexplained and requires comparative studies of peripheral and central parts of the thermoafferent system.

Animals↗
PubMedDate not supplied

Activation and suppression of the trapezius muscle induced by transcranial magnetic stimulation.

Motor evoked potentials (MEPs) and silent periods (SPs) in the trapezius muscle induced by transcranial magnetic stimulation (TMS) were investigated in 15 healthy subjects. Stimuli were applied with a Novametrix Magnetic stimulator using a 14 cm circular coil 4 cm lateral to the vertex on the biauricular line. Surface electrodes were used for simultaneous bilateral electromyographic recordings of the trapezius. TMS invariably induced contralateral MEPs (latency 10.5 +/- 1.3 ms, mean +/- SD), with ipsilateral responses in 53% of the subjects (latency 11.1 +/- 2.5 ms). The mean duration of the SPs was approximately 90 ms on both sides. There were no significant side differences between any of the MEP or SP parameters. To study the influence of subcortical inhibition phenomena TMS induced responses were assessed following electrical mental nerve stimulation with interstimulus intervals (ISI) of 0-100 ms. MEP latencies significantly increased at ISI of 10-100 ms, whereas MEP amplitudes and SPs did not change. These findings may reflect a trigeminal induced exteroceptive suppression of trapezius muscle activity.

Accessory Nerve↗