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H Breer

Publications and source records attributed to H Breer.

At least 127 records · Page 7Linked to original sources
PubMed1992

Expression of odorant receptors in spatially restricted subsets of chemosensory neurones.

From a rat olfactory library a cDNA clone (OR37) which is supposed to encode an odorant receptor protein has been isolated and characterized. Specific antisense RNA and in situ hybridization techniques have been employed to monitor the olfactory epithelium for the distribution of olfactory neurones expressing the OR37-gene. The OR37-transcripts were detected only in a subset of receptor cells segregated in two restricted areas of the olfactory epithelium. The clusters of reactive cells appear symmetrically in both nasal cavities. Within a reactive region only a subset of the cells expressed the receptor. The segregation of neurones expressing a distinct receptor supports the notion that a spatial component may be involved in coding odour quality.

Amino Acid Sequence↗
PubMed1991

Purification and reconstitution of the high affinity choline transporter.

The high-affinity choline transporter has been solubilized from synaptosomal membranes by various detergents. The solubilized carrier protein has been incorporated into liposomes after removal of the detergent by dialysis. Using the reconstitution of choline transport activity as an assay, the components catalyzing choline translocation were purified from the detergent extract by ion-exchange chromatography on a Mono-Q column followed by immunoaffinity chromatography. Monitoring the active fractions by sodium dodecylsulfate polyacrylamide gel electrophoresis and isoelectrofocussing gave one major protein with an apparent molecular weight of about 90,000 and an isoelectric point of pH 4.7. The isolated protein appeared to be heavily glycosylated as shown by lectin binding; upon treatment with endoglycosidase F the polypeptide was degraded to an apparent molecular weight of about 65,000. Accumulation of choline into liposomes reconstituted with the purified protein was driven by artificially imposed sodium gradients and inhibited by hemicholinium-3.

Animals↗
PubMed1991

Cloning of genomic and complementary DNA encoding insect pheromone binding proteins: evidence for microdiversity.

Genomic DNA from the silk moth Antheraea pernyi bearing the gene of a pheromone binding protein has been isolated from a partial genomic library using specific cDNA probes. The DNA spans 3.5 kilobases, contains three exons and two intervening sequences that interrupt the protein coding region of the gene. A DNA fragment of a second gene was isolated and the complete primary structure of a corresponding cDNA clone was unravelled. The expression of two different genes, giving rise to different pheromone binding proteins, implies a more specific function of these proteins than was hitherto assumed.

Amino Acid Sequence↗
PubMed1991

Generation of monoclonal antibodies detecting specific epitopes in olfactory and respiratory epithelia.

Two panels of monoclonal antibodies have been generated, each panel having a distinct specificity for antigens located in the ciliary zone of either the olfactory or respiratory epithelium of rats. Tissue specificity was confirmed in enzyme-linked immunosorbent assays on membrane fractions from various tissues. During ontogeny, the expression of olfactory-specific antigens preceeds that of respiratory-specific antigens; this observation correlates with differences in the genesis of the respective cilia type and confirms that different molecular entities are recognized. A spatial segregation of immunoreactivity in the chemosensory epithelium was observed for one of the olfactory-specific monoclonal antibodies; negative zones were located in the dorsal recess of the nasal cavity and on the tips of the turbinates. Olfactory-specific antibodies reacted with distinct polypeptide bands on Western blots from olfactory ciliary preparations.

Animals↗
PubMed1991

Molecular reaction cascades in olfactory signal transduction.

Odorant induced second messenger signals in ciliary preparations from rat olfactory epithelia were monitored in the subsecond time range using a rapid kinetic methodology. Application of micromolar concentrations of odorants induced a rapid and transient elevation of second messenger concentrations. The odorous compounds analyzed induced in a mutually exclusive way the formation of either cyclic adenosine monophosphate or inositol-triphosphate. The activating effects of odorants on intracellular signalling cascades appear to be mediated via different G-proteins. Thus, at least two different second messenger pathways appear to be involved in olfactory signal transduction. Selective inhibition of odor-induced second messenger responses by certain lectins indicate that glycoproteins appear to be involved in the perception or transduction of olfactory signals. In the presence of protein kinase inhibitors the odorant-induced second messenger response is no longer transient but persistent over a longer time period, suggesting that termination of the signal is realized via feedback phosphorylation of functional elements in the reaction cascade.

Adenylyl Cyclases↗
PubMed1990

Rapid kinetics of second messenger formation in olfactory transduction.

Olfactory transduction is thought to be mediated by a membrane-bound receptor protein initiating a multistep reaction cascade which ultimately leads to a depolarizing generator current. There is considerable evidence for the involvement of adenylate cyclase in vertebrate olfactory transduction, and some data indicate that phospholipase C may have a central role in insect olfaction. However, one must show that odorants not only stimulate enzyme activity but also induce changes in concentrations of relevant second messengers. One important criterion for a candidate second messenger of chemo-electrical transduction is that its formation must precede the onset of the odorant-induced membrane permeability changes which proceed on a subsecond time-scale. Here we report an odorant-induced, transient accumulation of cyclic AMP in isolated olfactory cilia from rats, and the generation of inositol trisphosphate in antennal preparations from insects, both of which show subsecond time courses that are sufficiently rapid to mediate the odorant-regulated permeability of olfactory receptor cells.

Animals↗
PubMed1990

Rapid activation of alternative second messenger pathways in olfactory cilia from rats by different odorants.

The molecular mechanisms mediating the chemo-electrical signal transduction in olfactory receptor cells are still elusive. In this study odor induced formation of second messengers in rat olfactory cilia was monitored in a subsecond time range using a rapid kinetic device. Application of micromolar concentration of citralva induced a rapid, transient elevation of the cyclic adenosine monophosphate level, whereas the concentration of inositol trisphosphate was not affected. In contrast, pyrazine caused a rise in the concentration of inositol trisphosphate, not affecting the level of cyclic adenosine monophosphate. Analysis of the kinetic parameter for the odorant induced reaction indicated that apparently two systems are operating simultaneously. The activating effects of odorants appear to be mediated via different G-proteins. Thus, at least two different second messenger pathways appear to be involved in olfactory signal transduction.

Animals↗
PubMed1990

Regulation of high affinity choline uptake.

High affinity uptake of choline, the rate-limiting, regulatory step for the synthesis of acetylcholine (ACh), was found to be regulated via presynaptic auto- and heteroreceptors. The transport rate was reduced by a muscarinic agonist and neuropeptides, but was significantly enhanced by octopamine. Intracellular messengers, including cyclic nucleotides, appear to modulate the transport activity, apparently by activating specific protein kinases.

Acetylcholine↗
PubMed1990

Identification of a cationic channel in synaptosomal membranes.

Synaptosomal membranes were fused with liposomes using the 'hydration technique' to produce giant proteoliposomes amenable to patch clamp recordings. Single channel currents of a cationic channel with particular properties were detected. In a solution of 150 mM NaCl, the channel displayed a unit conductance of 136 pS and a mean open state lifetime of 1.1 ms. The gating of the channel was shown to be voltage as well as calcium dependent. Pharmacological studies revealed that the channel was insensitive to a variety of channel blockers, but was inactivated by ruthenium red. Presumably, this channel may play a role in regulating the evoked release of neurotransmitters.

Animals↗
PubMed1990

Neuronal acetylcholine receptor channels from insects: a comparative electrophysiological study.

The channel properties of nicotinic acetylcholine receptor subtypes in the nervous system of insects (Locusta migratoria) have been characterized. Single channel measurements were performed using patch-clamp techniques as well as planar lipid bilayer reconstitution approaches. In reconstitution experiments using receptor-preparations isolated from neuronal membranes by alpha-toxin affinity chromatography, a ligand-gated channel type was found, which showed a high conductance and a short mean lifetime. Patch-clamp experiments on synapse-free somata of isolated nerve cells revealed an acetylcholine-gated channel type with a lower conductance but a longer lifetime. The two different agonist-activated channel types are supposed to represent synaptic and extrasynaptic acetylcholine receptors.

Animals↗
PubMed1990

Primary structure of a pheromone-binding protein from Antheraea pernyi: homologies with other ligand-carrying proteins.

An antennal cDNA clone encoding the complete sequence (163 amino acids) of a pheromone-binding protein precursor from the male silk moth, Antheraea pernyi, was isolated using oligonucleotide probes. The cloned cDNA was expressed and the translation product detected by specific antibodies. The deduced protein sequence consists of a signal peptide of 21 amino acids and a mature binding protein of 142 amino acid residues. The predicted structure of this protein is homologous to binding-proteins from different insect species which have previously been identified, but shows no similarities to odorant-binding proteins from vertebrates, suggesting that soluble odorant-binding proteins in insects and vertebrates represent an evolutionary convergence.

Amino Acid Sequence↗
PubMed1990

Odorant-sensitive phospholipase C in insect antennae.

Exogenous tritiated phosphatidylinositol bisphosphate added to antennal preparations from locust and cockroach was hydrolysed releasing inositol trisphosphate. High activity of phospholipase C was detected in the soluble as well as in the membrane fraction. At low free calcium concentrations hydrolysis of the labelled lipid was stimulated by odorants and pheromones in a GTP-dependent manner. Consequently the level of inositol trisphosphate in antennal preparations increased upon odorant stimulation.

Animals↗
PubMed1990

Molecular cloning, sequencing and expression of cDNA encoding a G0-protein from insect.

A locust cDNA clone encoding the complete sequence of a guanine nucleotide-binding protein was isolated and its nucleotide sequence determined. Comparing the deduced amino acid sequence with primary structures of other G-proteins revealed striking homologies with the vertebrate G0-protein. The cloned cDNA was expressed and the translation product detected by specific antibodies. Northern blot analysis revealed that the corresponding mRNA exists in two forms, preferentially expressed in the nervous tissue.

Amino Acid Sequence↗
PubMed1989

Immunocytochemical localization of nicotinic acetylcholine receptors in the terminal abdominal ganglion of the cockroach (Periplaneta americana).

A polyclonal, monospecific antiserum raised against a nicotinic acetylcholine receptor protein affinity-purified from insect nervous tissue, was employed to demonstrate the localization of antigenic sites in the neuropile of the terminal (sixth) abdominal ganglion of the cockroach Periplaneta americana. In agreement with previously published autoradiographic mapping of specific [125I]alpha-bungarotoxin binding sites, specific areas of the central neuropile of this ganglion were densely stained, but not the cercal afferent axons. No staining was detected corresponding to the dense, peripheral, partly non-specific binding of alpha-bungarotoxin seen in autoradiographs of the same tissue. Certain peripherally located neuronal cell bodies, including the cell body of giant interneuron 2, contained intracellularly located antigenic sites.

Animals↗
PubMed1989

Molecular cloning of an insect pheromone-binding protein.

Clones coding for the pheromone binding protein precursor have been selected from a cDNA library derived from antennae of the male moth, Antheraea polyphemus. The deduced protein sequence consists of a signal peptide of 20 amino acid residues and a mature binding protein of 142 amino acid residues. RNA blot hybridization indicated that the mRNA is selectively expressed in the antennae of the male moth.

Animals↗
PubMed1989

Isolation and reconstitution of the high-affinity choline carrier.

Monoclonal antibodies, which block the high-affinity uptake of choline in synaptosomal ghosts, have been used to purify a membrane polypeptide (80 kDa) from insect synaptosomal membranes. This isolated protein was found to catalyse the sodium-dependent, hemicholinium-sensitive accumulation of choline after reconstitution into liposomes, thus, apparently represents the high-affinity choline transporter.

Animals↗