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H Betz

Publications and source records attributed to H Betz.

At least 217 records · Page 12Linked to original sources
PubMed1988

Characterization of an invertebrate nicotinic acetylcholine receptor gene: the ard gene of Drosophila melanogaster.

The ard gene encodes a neuronal nicotinic acetylcholine receptor (AChR) protein from Drosophila (ARD protein). Cytogenetically this gene maps at position 64B/C on the left arm of the 3rd chromosome. Five introns interrupt the protein coding region of the gene, and one is found upstream of the translation start site. The ard gene thus contains less introns than vertebrate muscle AChR genes, but, with one exception, the positions of the resident introns are precisely conserved. Implications for the evolution of AChR genes are discussed.

Animals↗
PubMed1988

Inhibition of beta-bungarotoxin binding to brain membranes by mast cell degranulating peptide, toxin I, and ethylene glycol bis (beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid.

The presynaptically active snake venom neurotoxin beta-bungarotoxin (beta-Butx) is known to affect neurotransmitter release by binding to a subtype of voltage-activated K+ channels. Here we show that mast cell degranulating (MCD) peptide from bee venom inhibits the binding of 125I-labeled beta-Butx to chick and rat brain membranes with apparent Ki values of 180 nM and 1100 nM, respectively. The mechanism of inhibition by MCD peptide is noncompetitive, as is inhibition of 125I-beta-Butx binding by the protease inhibitor homologue from mamba venom, toxin I. Beta-Butx and its binding antagonists thus bind to different sites of the same membrane protein. Removal of Ca2+ by ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid inhibits the binding of 125I-beta-Butx by lowering its affinity to brain membranes.

Animals↗
PubMed1988

Neuronal acetylcholine receptors in Drosophila: the ARD protein is a component of a high-affinity alpha-bungarotoxin binding complex.

The ard gene of Drosophila melanogaster encodes a structural homologue of vertebrate nicotinic acetylcholine receptors (AChR) and is expressed exclusively in nervous tissue. To study the nature of the ARD protein, antibodies were raised against fusion constructs containing two regions of this polypeptide. One segment is putatively extracellular (amino acids 65-212), the other domain is exposed to the cytoplasm (amino acids 305-444). The ARD antisera obtained served to investigate the physical relationship between the ARD protein and alpha-bungarotoxin (alpha-Btx) binding sites occurring in Drosophila. Two different high-affinity binding sites for [125I]alpha-Btx, a highly potent antagonist of vertebrate muscle AChR, were detected in fly head membranes. Equilibrium binding and kinetic studies revealed Kd values of approximately 0.1 nM (site 1) and approximately 4 nM (site 2). The estimated maximal binding (Bmax) was approximately 240 and 1080 fmol/mg protein respectively. Both sites exhibited a nicotinic-cholinergic pharmacology. Immunoprecipitation experiments with the ARD antisera indicated that the ARD protein is associated with the [125I]alpha-Btx binding site 1 only. These data support the previously postulated hypothesis that the ARD protein is part of an alpha-Btx binding neuronal AChR of Drosophila. Furthermore, they indicate heterogeneity in nicotinic-cholinergic binding sites in the insect nervous system.

Animals↗
PubMed1988

Basilar artery disease--clinical outcome and Doppler sonographical follow-up.

In the past 5 years we have investigated 29 patients with symptomatic basilar artery stenoses (14 cases) and occlusions (14) and a patent primitive trigeminal artery with thin-calibered basilar and vertebral arteries (1) using directional continuous-wave Doppler sonography of the vertebral arteries. A total of 19 patients survived, and 17 of them were clinically and sonographically reexamined after 40.4 +/- 15.8 months (mean +/- SD). Among the 8 patients with basilar stenoses, 6--with no further transient ischemic attacks (TIAs) in the interval--exhibited an increase in the summed modified Pourcelot indices (relative end-diastolic flow velocities) of the vertebrals by 0.18 +/- 0.16; the other 2 showed a decrease by 0.26 each, in 1 case temporally related to a TIA, in the 2nd case without further clinical deterioration. In the 8 survivors with basilar occlusions, 5 remained--by sonographic criteria--unchanged with summed modified Pourcelot indices of the vertebrals of 0.00, while 3 patients exhibited a slight increase in the summed modified Pourcelot indices of 0.13 +/- 0.03. While the difference between the outcome of subsets of patients treated with regimens of 30,000-40,000 units heparin/day or phenprocoumon and less radical drugs were statistically not significant, the former regimen appeared clinically more efficacious in preventing further deterioration in approximately two-thirds of the patients affected. Due to the potential recurrence of neurological symptoms, a treatment period with phenprocoumon of 6 months after discharge from hospital appears justified. Due to these therapeutic efforts, approximately half of the patients initially affected survived with no or only a mild neurological deficit.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗
PubMed1988

The efficacy of EC-IC anastomoses: evaluation by directional c-w Doppler sonography.

We report the results obtained by means of directional c-w Doppler sonography on 33 patients with superficial temporal-to-middle cerebral artery anastomoses. The efficiency of the anastomosis was evaluated by the modified Pourcelot indices (relative end-diastolic flow velocities) of the preauricular superficial temporal artery and of the bypass-supplying branch at the edge of the burr-hole. The influence of intermittent compression of the bypass-supplying branch on the modified Pourcelot index of the ipsilateral common carotid was used as a further criterion in the sonographic evaluation. All efficient anastomoses, defined by a modified Pourcelot index of at least 0.20 at the edge of the burr-hole, exhibited a reduction of the relative end-diastolic flow velocity of 0.08 on the average in the common carotid during compression. In the 18 cases with unilateral occlusion of the internal carotid, bypass surgery was predominantly efficacious in those patients who showed a reduction in the sum of the modified Pourcelot indices of the remaining brain-supplying arteries of at least two standard deviations with respect to the mean of age-matched controls. The subgroups of patent and absent collaterals through the ophthalmic artery did not show any difference with respect to the percentage of efficient anastomoses. In all 4 patients with bilateral internal carotid artery occlusion, bypass surgery was effective, while 50% of the patients with intracranial carotid artery disease exhibited an insufficient anastomosis.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1988

Localization of glycine receptors in the rat central nervous system: an immunocytochemical analysis using monoclonal antibody.

The localization of glycine receptors was immunocytochemically examined in the rat brain using a monoclonal antibody against the affinity-purified glycine receptor. Glycine receptors were concentrated in the lower brainstem, whereas no immunoreactivity was observed in the diencephalon and forebrain except in a few diencephalic nuclei. The highest density of receptors was found in the cranial motor nuclei, reticular formation, parabrachial area, dorsal and ventral cochlear nuclei, and dorsal and ventral tegmental nuclei. Differences were observed in the distribution of immunoreactive elements in the various brain regions. In the cerebellar cortex, the immunoreactivity was exclusively seen along the dendrites of the Purkinje cells. On the other hand, glycine receptors were detected on the cellular membrane of the soma of the cochlear nuclei, trigeminal motor nucleus, parabrachial area, lateral reticular nucleus, dorsal nucleus of the lateral lemniscus, cerebellar nuclei, trigeminal spinal nucleus, anterior horn and reticular formation. In other regions, the receptors were evenly distributed throughout the neuropil.

Animals↗
PubMed1988

Conserved quaternary structure of ligand-gated ion channels: the postsynaptic glycine receptor is a pentamer.

The postsynaptic glycine receptor of rat spinal cord is a glycosylated membrane protein that, after affinity purification, contains membrane-spanning subunits of Mr 48,000 and 58,000 and an associated peripheral polypeptide of Mr 93,000. Here, the quaternary structure of the transmembrane core of the receptor was investigated by chemically crosslinking its subunits. Upon treatment with crosslinking reagents of different side-chain specificities and lengths, a consistent set of adducts up to Mr 260,000 was detected after separation by NaDodSO4/PAGE. The observed pattern of adducts was similar irrespective of whether purified receptor protein or synaptosomal membranes were crosslinked. Compositional analysis revealed that the crosslinked adducts contained the Mr 48,000 and 58,000 subunits in varying ratios but not the peripheral Mr 93,000 polypeptide. Thus adducts of intermediate molecular weight represent dimers, trimers, and tetramers of the transmembrane subunits, whereas the major adduct of Mr 260,000 corresponds to a pentameric assembly of subunits forming the ion channel of the glycine receptor. This subunit arrangement is similar to that reported for the nicotinic acetylcholine receptor of fish electric organ and skeletal muscle. Hence, we suggest that the different ligand-gated ion channels of excitable membranes share a similar quaternary structure.

Animals↗
PubMed1988

Molecular cloning of the antagonist-binding subunit of the glycine receptor.

The postsynaptic receptor for the inhibitory neurotransmitter glycine is a heterooligomeric membrane protein which, after affinity purification on an antagonist column, contains three polypeptides of 48K, 58K and 93K. Sequencing of cDNA clones of the antagonist-binding 48K subunit revealed a structural organization similar to and significant amino acid homology with nicotinic acetylcholine receptor proteins. Our data suggest the existence of a set of related genes encoding transmembrane channel-forming neurotransmitter receptor polypeptides of excitable membranes.

Amino Acid Sequence↗
PubMed1987

Co-localization of choline acetyltransferase and postsynaptic glycine receptors in motoneurons of rat spinal cord demonstrated by immunocytochemistry.

Male rats were perfused with paraformaldehyde and picric acid. The cervical spinal cord was cryosectioned and immunostained with a monoclonal antibody against the postsynaptic receptor for the neurotransmitter glycine. The anterior horn contained glycinoceptive neurons of varying morphology. Cholinergic cells were identified in the same tissue sections when subsequently immunostained with a monoclonal antibody against choline acetyltransferase, the biosynthetic enzyme of acetylcholine. Immunoreactivity for the glycine receptor was detected in the plasma membrane and for choline acetyltransferase in the perikaryal cytoplasm of identical anterior horn cells, classified as small, medium and large motoneurons. This suggests that motoneurons have receptors for glycine on their cell surface.

Animals↗
PubMed1987

The chloride channel blocking agent, t-butyl bicyclophosphorothionate, binds to the gamma-aminobutyric acid-benzodiazepine, but not to the glycine receptor in rodents.

The inhibitory neurotransmitters glycine and gamma-aminobutyric acid (GABA) both activate transmembrane chloride channels of similar physical characteristics. A common ion channel component has therefore been postulated for both the glycine and GABA receptor proteins. Different convulsant drugs as picrotoxin and t-butyl bicyclophosphorothionate (TBPS) have been reported as channel-blocking ligands of the GABA receptor. Here, we show that the distribution of [35S]TBPS binding sites parallels the binding of the GABA receptor ligand [3H]flunitrazepam, but not that of the glycine receptor antagonist [3H]strychnine. Binding was examined in membrane fractions from different regions of the rat CNS and of the mutant mouse spastic, an animal deficient in glycine receptors. Also, affinity purification of the glycine receptor on aminostrychnine-agarose resulted in almost complete removal of [35S]TPBS binding sites from the receptor preparation. It is concluded that TBPS selectively binds to the GABA, but not glycine, receptor chloride channel complex.

Animals↗
PubMed1987

The Mr 93,000 polypeptide of the postsynaptic glycine receptor complex is a peripheral membrane protein.

The glycine receptor of mammalian spinal cord is an oligomeric membrane protein that, after affinity purification on aminostrychnine-agarose or immobilized antibody, contains three polypeptides of Mr 48,000, 58,000, and 93,000. Here, the association and the properties of the polypeptides of the rat glycine receptor were investigated. Upon phase partitioning in the nonionic detergent Triton X-114, the three receptor polypeptides behaved as a hydrophilic protein complex exhibiting phospholipid binding. Sucrose gradient centrifugation or gel filtration in the presence of dithiothreitol and Triton X-100 separated the Mr 93,000 polypeptide from the Mr 48,000 and 58,000 polypeptides, which harbor the antagonist binding site of the glycine receptor. Alkaline or dimethylmaleic acid anhydride treatment of crude synaptic membrane fractions resulted in extraction of the Mr 93,000 polypeptide. Lectin binding was observed for the Mr 48,000 and 58,000 glycine receptor subunits but not the Mr 93,000 polypeptide. These results indicate that the Mr 93,000 polypeptide is a peripheral membrane protein that is located at the cytoplasmic face of the postsynaptic glycine receptor complex.

Amino Acids↗
PubMed1987

Synaptophysin: molecular organization and mRNA expression as determined from cloned cDNA.

Synaptophysin is a major glycoprotein of Mr approximately 38,000 (in deglycosylated form: Mr approximately 34,000) characteristic of a certain class of small (30-80 nm diameter) neurosecretory vesicles, including presynaptic vesicles, but also vesicles of various neuroendocrine cells of both neuronal and epithelial phenotype. Using synaptophysin-specific antibodies we have isolated cDNA clones from rat nervous tissue libraries, which identify an approximately 2.5-kb mRNA in rat and human cells, including neuroendocrine tumours, that contains a reading frame for a polypeptide of 307 amino acids with a total mol. wt of 33 312. The deduced amino acid sequence, which was partly confirmed by comparison with sequences of two tryptic peptides obtained from purified synaptophysin, revealed four hydrophobic regions of 24 amino acids each, which are characterized, according to conformation prediction analyses, by marked alpha-helicity. The sequence shows a single potential N-glycosylation site, which is assigned to the vesicle interior, and a carboxy-terminal tail of 89 amino acids which contains glycine-rich tetrapeptide repeats, the epitope of monoclonal antibody SY38, and a number of collagenase-sensitive sites accessible on the surface of the intact vesicles. These features suggest that the polypeptide spans the vesicle membrane four times, with both N and C termini located on the outer, i.e. cytoplasmic, surface of the vesicles.

Amino Acid Sequence↗
PubMed1987

Vertebral artery Doppler sonography.

We have examined the vertebral and subclavian arteries in 1,205 patients using directional continuous-wave (c-w) Doppler sonography, and compared the sonographic findings with the results of unilateral or bilateral retrograde brachial arteriographies in the same patients. Doppler sonography revealed 33 false positives among 909 cases with normal angiographic findings. Some types of vertebral artery (VA) lesions allowed an excellent, others a fairly good differentiation by Doppler sonography: the complete subclavian steal syndrome with constant reversal of VA flow was reliably detected (16 cases). In the incomplete steal syndrome (5 cases) sonography was superior to angiography. Two bilateral distal VA occlusions and seven basilar artery occlusions - six in the proximal third and one in the rostral third - were detected sonographically; four basilar occlusions sparing the caudal third and one case exhibiting rete mirabile anastomoses were not identified by Doppler sonography. Our acoustically defined sonographic criteria did not permit an unequivocal assignment to an anatomical variant or a vascular lesion. The sensitivity in the detection of a severe stenosis at the VA origin amounted to 16 out of 31, and to 12 of 25 in cases with a proximal VA occlusion and reconstitution of the distal VA through cervical collaterals. Our results confirm that the conventional hand-held c-w Doppler sonography cannot replace angiography in the evaluation of vertebro-basilar insufficiency. It rather serves as an aid to the decision for or against angiography, and in the follow-up of angiographically proven lesions. However, several therapeutically important lesions are readily diagnosed by sonography.

Angiography↗
PubMed1987

Identification of gene products expressed in the developing chick visual system: characterization of a middle-molecular-weight neurofilament cDNA.

A cDNA library enriched for transcripts that accumulate from embryonic day 7 until day 1 posthatching was constructed from poly(A)+ RNA of chick optic lobe tissue. From this library, three recombinants were isolated that correspond to neural mRNAs appearing during the major period of synaptogenesis in the retinotectal system. One of these recombinants (OZ 11) was identified as a middle-molecular weight neurofilament (NF-M) cDNA which, together with two overlapping clones, encodes part of the rod and the entire tail region of NF-M. In situ hybridization revealed NF-M mRNA to be highly expressed in regions of the chick central nervous system, which contain multipolar and/or long-projection neurons.

Amino Acid Sequence↗
PubMed1986

Identification of glycinergic synapses in the cochlear nucleus through immunocytochemical localization of the postsynaptic receptor.

The distribution and morphology of glycinergic synapses in the cochlear nucleus were investigated using monoclonal antibodies against the glycine receptor. Glycine receptor immunoreactivity was seen on somas and proximal processes of most cells in all divisions of the cochlear nucleus; distribution of label in neuropil was denser in the dorsal cochlear nucleus and granule cell cap than in the ventral cochlear nucleus. At the ultrastructural level, glycine receptor immunoreactivity was specifically distributed postsynaptically to terminals that contained flattened vesicles in the guinea pig anteroventral cochlear nucleus. These studies show that the immunocytochemical localization of the glycine receptor can provide a means of identifying and characterizing glycinergic synapses throughout the central nervous system.

Animals↗
PubMed1986

Molecular characterization of synaptophysin, a major calcium-binding protein of the synaptic vesicle membrane.

Synaptophysin, a mol. wt 38 000 glycopolypeptide of the synaptic vesicle membrane, was solubilized using Triton X-100 and purified by immunoaffinity or ion-exchange chromatography. From gel permeation and sucrose-density centrifugation in H2O/D2O, a Stokes radius of 7.3 nm, a partial specific volume of 0.830 and a total mol. wt of 119 000 were calculated for the native protein. Cross-linking of synaptic vesicles with glutaraldehyde, dimethylsuberimidate, or Cu2+ -o-phenantroline, resulted in the formation of a mol. wt 76 kd dimer of synaptophysin. Crosslinking of the purified protein in addition produced tri- and tetrameric adducts of the polypeptide. Native synaptophysin thus is a homooligomeric protein. Synaptophysin is N-glycosylated, since cultivation of the rat phaeochromocytoma cell line PC12 in the presence of tunicamycin reduced its mol. wt by about 6 kd. Upon transfer to nitrocellulose and incubation with 45Ca2+, synaptophysin behaved as one of the major calcium-binding proteins of the synaptic vesicle membrane. Pronase treatment of intact synaptic vesicles abolished this 45Ca2+ binding indicating that the Ca2+ binding site of synaptophysin must reside on a cytoplasmic domain of the transmembrane polypeptide. Based on these data, we propose that synaptophysin may play an important role in Ca2+-dependent neurotransmitter release.

Animals↗