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R Jahn

Publications and source records attributed to R Jahn.

At least 181 records · Page 10Linked to original sources
PubMed1989

A synaptic vesicle membrane protein is conserved from mammals to Drosophila.

The structure of synaptobrevin, an intrinsic membrane protein of small synaptic vesicles from mammalian brain, was studied by purification and molecular cloning. Its message in bovine brain encodes a 116 amino acid protein whose sequence reveals it to be the mammalian homolog of Torpedo VAMP-1. Antibody probing demonstrates that the protein is also present in Drosophila, and its Drosophila homolog was cloned. Alignment of the sequences of synaptobrevin/VAMP-1 from the three species shows it to contain four domains, including a highly conserved central region of 63 amino acids that contains 75% invariant residues. The finding that a membrane protein from vertebrate synaptic vesicles is conserved in Drosophila points toward a central role of this protein in neurotransmission and should allow a genetic approach to neurotransmitter release.

Amino Acid Sequence↗
PubMed1989

Synaptic vesicles immunoisolated from rat cerebral cortex contain high levels of glutamate.

L-Glutamate is regarded as the major excitatory neurotransmitter in the mammalian CNS. However, whether the released transmitter originates from a cytosolic pool or is discharged from synaptic vesicles by exocytosis (vesicle hypothesis) remains controversial. A problem with the general acceptance of the vesicle hypothesis is that the enrichment of glutamate in synaptic vesicles has not been convincingly demonstrated. In the present study, we have analyzed the glutamate content of synaptic vesicles isolated from rat cerebral cortex by a novel immunobead procedure. A large amount of glutamate was present in these vesicles when a proton electrochemical gradient was maintained across the vesicle membrane during isolation. Compared with the starting fraction, glutamate was enriched more than 10-fold relative to other amino acids. Addition of N-ethylmaleimide prevented glutamate loss during isolation. Isotope exchange experiments revealed that exchange or re-uptake of glutamate after homogenization is negligible. We conclude that rat brain synaptic vesicles contain high levels of glutamate in situ.

Animals↗
PubMed1989

Interactions of synapsin I with small synaptic vesicles: distinct sites in synapsin I bind to vesicle phospholipids and vesicle proteins.

Synapsin I is a major neuron-specific phosphoprotein that is specifically localized to the cytoplasmic surface of small synaptic vesicles. In the present study, the binding of synapsin I to small synaptic vesicles was characterized in detail. The binding of synapsin I was preserved when synaptic vesicles were solubilized and reconstituted in phosphatidylcholine. After separation of the protein and lipid components of synaptic vesicles under nondenaturing conditions, synapsin I bound to both components. The use of hydrophobic labeling procedures allowed the assessment of interactions between phospholipids and synapsin I in intact synaptic vesicles. Hydrophobic photolabeling followed by cysteine-specific cleavage of synapsin I demonstrated that the head domain of synapsin I penetrates into the hydrophobic core of the bilayer. The purified NH2-terminal fragment, derived from the head domain by cysteine-specific cleavage, bound to synaptic vesicles with high affinity confirming the results obtained from hydrophobic photolabeling. Synapsin I binding to synaptic vesicles could be inhibited by the entire molecule or by the combined presence of the NH2-terminal and tail fragments, but not by an excess of either NH2-terminal or tail fragment alone. The purified tail fragment bound with relatively high affinity to synaptic vesicles, though it did not significantly interact with phospholipids. Binding of the tail fragment was competed by holosynapsin I; was greatly decreased by phosphorylation; and was abolished by high ionic strength conditions or protease treatment of synaptic vesicles. The data suggest the existence of two sites of interaction between synapsin I and small synaptic vesicles: binding of the head domain to vesicle phospholipids and of the tail domain to a protein component of the vesicle membrane. The latter interaction is apparently responsible for the salt and phosphorylation dependency of synapsin I binding to small synaptic vesicles.

Animals↗
PubMed1989

Microvesicles of the neurohypophysis are biochemically related to small synaptic vesicles of presynaptic nerve terminals.

Nerve endings of the posterior pituitary are densely populated by dense-core neurosecretory granules which are the storage sites for peptide neurohormones. In addition, they contain numerous clear microvesicles which are the same size as small synaptic vesicles of typical presynaptic nerve terminals. Several of the major proteins of small synaptic vesicles of presynaptic nerve terminals are present at high concentration in the posterior pituitary. We have now investigated the subcellular localization of such proteins. By immunogold electron microscopy carried out on bovine neurohypophysis we have found that three of these proteins, synapsin I, Protein III, and synaptophysin (protein p38) were concentrated on microvesicles but were not detectable in the membranes of neurosecretory granules. In addition, we have studied the distribution of the same proteins and of the synaptic vesicle protein p65 in subcellular fractions of bovine posterior pituitaries obtained by sucrose density centrifugation. We have found that the intrinsic membrane proteins synaptophysin and p65 had an identical distribution and were restricted to low density fractions of the gradient which contained numerous clear microvesicles with a size range the same as that of small synaptic vesicles. The peripheral membrane proteins synapsin I and Protein III exhibited a broader distribution extending into the denser part of the gradient. However, the amount of these proteins clearly declined in the fractions preceding the peak of neurosecretory granules. Our results suggest that microvesicles of the neurohypophysis are biochemically related to small synaptic vesicles of all other nerve terminals and argue against the hypothesis that such vesicles represent an endocytic byproduct of exocytosis of neurosecretory granules.

Animals↗
PubMed1989

[Resorbable implants and their use exemplified by fracture of the head of the radius].

First a short survey of the variety of experiments concerning absorbable implants is given. The result of those experiments, especially with large implants are still quite unsatisfactory. For small osteosynthesis the development of Polydioxanon-Pins seems to open new perspectives of therapy. Also some selected indications in the field of hand surgery as well as maxillary surgery are mentioned. Promising characteristics of PDS-Pins, such as gentle application, complete absorbability, no late infections and no removal of metal plates, made us implant these pins in eight patients aged 35 to 58 suffering from radial head fractures (type Ib and Ic according to Vogt). Early functional treatment showed very good results. Two exemplary cases are illustrated.

Female↗
PubMed1989

[Indications for surgical management of humerus diaphyseal fractures and results].

Humerus fractures have continued to be associated with conservative and functional therapy. Indications for osteosynthesis may result either from failure of conservative treatment or from concomitant complications. Such cases are discussed in this paper. The patients with surgical treatment should not be considered as a group of "negative" selection, but osseous consolidation was accomplished in all of them. DC plate application has proved to be the optional method.

Adult↗
PubMed1989

[Damage to the arm plexus caused by atypical callus formation following clavicular fracture].

Secondary brachial plexus paralysis do not occur very often as a consequence of collar bone fractures. It is difficult to detect the cause, especially as there can be quite a long interval between the trauma and occurrence of the first symptoms. The problem is discussed with reference to a case observed by the authors: atypical callus formation 7 weeks after a clavicular fracture that was conservatively treated and had healed had led to brachial plexus damage. The patient was treated 11 months later by a trauma surgeon. Once the callus had been removed she was finally free of symptoms.

Adult↗
PubMed1989

[Personal experiences in the reconstruction of trauma-induced bone defects].

In this report an analysis of all patients treated with bone grafting in a 5-year period is given. Bone grafting was necessary in 8.4% of patients treated by osteosynthesis. Bone defects are seen mostly in the lower extremities. Malunion and nonunion are relatively frequent after fractures of the forearm and tibia. Contaminated fractures were seen in 32 patients. Before any osseous reconstruction, the defects of skin and soft tissues had to be closed and local infections treated. The period between accident and bone grafting ranged between 3 weeks and 1 1/2 years. Only in stage I fractures is primary grafting indicated. Intensive treatment of infection was required first in 16.9% of cases. When segmental defects are present, multistaged bone grafting entails only a low degree of risk and usually successful. In 87% autogenic cancellous iliac bone was transplanted. The use of allogenic spongiosa cannot be recommended for traumatic defects.

Bone Transplantation↗
PubMed1988

Glutamate uptake by brain synaptic vesicles. Energy dependence of transport and functional reconstitution in proteoliposomes.

The dependence of glutamate uptake on ATP-generated proton electrochemical potential was studied in a highly purified preparation of synaptic vesicles from rat brain. At low chloride concentration (4 mM), the proton pump present in synaptic vesicles generated a large membrane potential (inside-positive), associated with only minor acidification. Under these conditions, the rate of L-[3H]glutamate uptake was maximal. In addition, L-glutamate induced acidification of the vesicle interior. D-Glutamate produced only 40% of the effect, and L-aspartate or gamma-aminobutyric acid produced less than 5%. The initial rate of glutamate-induced acidification increased with increasing glutamate concentration. It was saturable and showed first-order kinetics (KM = 0.32 mM). Correspondingly, L-glutamate induced a small reduction in the membrane potential. The rate of ATP hydrolysis was unaffected. In comparison, glutamate had no effect on acidification or membrane potential in resealed membranes of chromaffin granules. At high chloride concentration (150 mM), the vesicular proton pump generated a large pH difference, associated with a small change in membrane potential. Under these conditions, uptake of L-[3H]glutamate by synaptic vesicles was low. For reconstitution, vesicle proteins were solubilized with the detergent sodium cholate, supplemented with brain phospholipids, and incorporated into liposomes. Proton pump and glutamate uptake activities of the proteoliposomes showed properties similar to those of intact vesicles indicating that the carrier was reconstituted in a functionally active form. It is concluded that glutamate uptake by synaptic vesicles is dependent on the membrane potential and that all components required for uptake are integral parts of the vesicle membrane.

Acridine Orange↗
PubMed1988

Uptake of GABA by rat brain synaptic vesicles isolated by a new procedure.

Uptake of GABA was demonstrated in rat brain synaptic vesicles which were prepared by a new and efficient procedure. The uptake activity co-purified with the synaptic vesicles during the isolation procedure. The purity of the vesicle fraction was rigorously examined by analysis of marker enzymes and marker proteins and also by immunogold electron microscopy using antibodies against p38 (synaptophysin). Contamination by other cellular components was negligible, indicating that GABA uptake by the synaptic vesicle fraction is specific for synaptic vesicles and not due to the presence of other structure possessing GABA uptake or binding activities. GABA uptake was ATP dependent and similar to the uptake of glutamate, which was assayed for a comparison. Both uptake activities were independent of sodium. They were inhibited by the uncoupler carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone, indicating that the energy for the uptake is provided by an electrochemical proton gradient. This gradient is generated by a proton ATPase of the vacuolar type as suggested by the effects of various ATPase inhibitors on neurotransmitter uptake and proton pumping. Competition experiments revealed that the transporters for GABA and glutamate are selective for the respective neurotransmitters.

Adenosine Triphosphatases↗
PubMed1988

Quantitation of nerve terminal populations: synaptic vesicle-associated proteins as markers for synaptic density in the rat neostriatum.

This study has assessed the contributions of the corticostriatal fibers, the ascending striatopetal fibers, and the intrinsic neostriatal neurons to the nerve terminal population found in the rat neostriatum (caudatoputamen). For this purpose, we have analysed the levels of two different synaptic vesicle-associated proteins, synapsin I and protein p38 (also called synaptophysin), in the neostriatum after specific lesions. Our results indicate that 45-50% of the synaptic vesicle proteins in the rat neostriatum derive from the corticostriatal fibers, that approximately 25-30% of the synaptic vesicle proteins are present in kainic acid-sensitive structures, presumably intrinsic terminals and local collaterals, and that ascending fibers contain 20-25% of the vesicle-associated proteins in the neostriatum. These three neuronal populations therefore comprise 95-100% of the synaptic vesicle-associated proteins in the rat neostriatum, and thus make up most of the nerve terminals in this brain region. The results, which are in general agreement with previous morphometric studies on the rat basal ganglia, therefore indicate that nerve terminals in the central nervous system can be quantitated by use of these biochemical nerve terminal markers. The results also indicate that a somewhat higher percentage of neostriatal nerve terminals belongs to the corticostriatal fibers that previously believed.

Animals↗
PubMed1988

Immunohistochemical and ultrastructural localisation of peptide-containing nerves and myocardial cells in the human atrial appendage.

The innervation and myocardial cells of the human atrial appendage were investigated by means of immunocytochemical and ultrastructural techniques using both tissue sections and whole mount preparations. A dense innervation of the myocardium, blood vessels and endocardium was revealed with antisera to general neuronal (protein gene product 9.5 and synaptophysin) and Schwann cell markers (S-100). The majority of nerve fibres possessed neuropeptide Y immunoreactivity and were found associated with myocardial cells, around small arteries and arterioles at the adventitial-medial border and forming a plexus in the endocardium. Subpopulations of nerve fibres displayed immunoreactivity for vasoactive intestinal polypeptide, somatostatin, substance P and calcitonin gene-related peptide. In whole-mount preparations of endocardium, substance P and calcitonin gene-related peptide immunoreactivities were found to coexist in the same varicose nerve terminals. Ultrastructural studies revealed the presence of numerous varicose terminals associated with myocardial, vascular smooth muscle and endothelial cells. Neuropeptide Y immunoreactivity was localised to large electron-dense secretory vesicles in nerve terminals which also contained numerous small vesicles. Atrial natriuretic peptide immunoreactivity occurred exclusively in myocardial cells where it was localised to large secretory vesicles. The human atrial appendage comprises a neuroendocrine complex of peptide-containing nerves and myocardial cells producing ANP.

Adult↗
PubMed1988

Localization of synapsin I at the frog neuromuscular junction.

We report here the results of immunocytochemical and biochemical studies on the localization of synapsin I, a nerve terminal--specific phosphoprotein, at the frog neuromuscular junction. Our results show that in this in situ synapse synapsin I is concentrated in the presynaptic compartment, where it appears to be associated with the synaptic vesicle membrane. Double immunoprecipitated synapsin I from homogenates of frog cutaneous pectoris muscles could be phosphorylated by the catalytic subunit of cyclic adenosine 5'-monophosphate-dependent protein kinase after gel electrophoresis and blotting onto nitrocellulose and could be subsequently identified by an immunoperoxidase technique. Experiments carried out in frog brain preparations indicate that frog synapsin I, like the mammalian protein, can be phosphorylated at different sites by exogenously added catalytic subunit of cyclic adenosine 5'-monophosphate-dependent protein kinase and Ca2+/calmodulin-dependent protein kinase II prepared from mammalian sources. The phosphorylation sites of frog synapsin I, as judged by phosphopeptide mapping, are somewhat different from those of mammalian synapsin I. The study of synapsin I and of the regulation of its state of phosphorylation at the neuromuscular junction may provide important information on its role in synaptic function, since at the present time this is one of the few systems in which a correlation among biochemical, immunocytochemical and electrophysiological results is possible.

Animals↗
PubMed1988

Synaptophysin (p38) at the frog neuromuscular junction: its incorporation into the axolemma and recycling after intense quantal secretion.

Recycling of synaptophysin (p38), a synaptic vesicle integral membrane protein, was studied by the use of antisera raised against the protein purified from frog brain. When frog cutaneous pectoris muscles were fixed at rest, a bright, specific immunofluorescent signal was observed in nerve-terminal regions only if their plasma membranes had been previously permeabilized. When muscles were fixed after they had been treated for 1 h with a low dose of alpha-latrotoxin in Ca2+-free medium, an equally intense fluorescence could be observed without previous permeabilization. Under this condition, alpha-latrotoxin depletes nerve terminals of their quantal store of acetylcholine and of synaptic vesicles. These results indicate that fusion of synaptic vesicles leads to the exposure of intravesicular antigenic determinants of synaptophysin on the outer surface of the axolemma, and provide direct support for the vesicle hypothesis of neurotransmitter release. After 1 h treatment with the same dose of alpha-latrotoxin in the presence of 1.8 mM extracellular Ca2+, immunofluorescent images were obtained only after permeabilization with detergents. Under this condition, the vesicle population was maintained by an active process of recycling and more than two times the initial store of quanta were secreted. Thus, despite the active turnover of synaptic vesicles and of quanta of neurotransmitter, no extensive intermixing occurs between components of the vesicle and presynaptic plasma membrane.

Animals↗
PubMed1988

The synaptic vesicle proteins synapsin I and synaptophysin (protein P38) are concentrated both in efferent and afferent nerve endings of the skeletal muscle.

Synapsin I and synaptophysin (protein p38) are 2 major protein components of the membranes of small synaptic vesicles of virtually all presynaptic nerve endings. Synapsin I, a phosphoprotein regulated by both Ca2+ and cAMP, is a peripheral protein of the cytoplasmic surface of the vesicle membrane. It is thought to anchor the vesicle surface to the cytoskeleton of the terminal and to play a regulatory role in neurotransmitter release. Synaptophysin is an intrinsic transmembrane glycoprotein. We report here that both proteins are present and concentrated also in afferent nerve endings, which provide the sensory innervation of the skeletal muscle and of the tendon. The distribution of both antigens in sensory nerve endings is consistent with their localization on the microvesicles that have been described in such endings. Thus, our results suggest the existence of important biochemical, and possibly functional, similarities between small synaptic vesicles of presynaptic nerve endings and microvesicles of sensory endings. Such findings provide new clues to the understanding of the physiology of sensory endings.

Afferent Pathways↗
PubMed1988

Secretory function of the vestibular nerve calyx suggested by presence of vesicles, synapsin I, and synaptophysin.

Type I sensory hair cells of the vestibular epithelium are nearly completely ensheathed by an afferent nerve ending, the vestibular nerve calyx. We have recently reported that the nerve calyx, and, in particular, its apical portions surrounding the neck of the hair cell, are immunoreactive for synapsin I (Favre et al., 1986), a major membrane component of small synaptic vesicles of axonal endings. We have now found, by electron microscopy, that the same region of the calyx is densely populated by microvesicles morphologically similar to typical presynaptic small synaptic vesicles. Furthermore, we have established by light microscopy immunocytochemistry that this region of the calyx also contains a high concentration of synaptophysin, another well-characterized major component of small synaptic vesicle membranes. These results suggest that the upper portion of the calyx is equipped with the machinery that in presynaptic terminals is involved in the release of neurotransmitters and raise the possibility that the calyx, via secretion of neurotransmitterlike substances, might modulate the function of type I hair cells.

Animals↗
PubMed1987

A synaptic vesicle protein with a novel cytoplasmic domain and four transmembrane regions.

Complementary DNA and genomic clones were isolated and sequenced corresponding to rat and human synaptophysin (p38), a major integral membrane protein of synaptic vesicles. The deduced amino acid sequences indicate an evolutionarily highly conserved protein that spans the membrane four times. Both amino and carboxyl termini face the cytoplasm, with the latter containing ten copies of a tyrosine-rich pentapeptide repeat. The structure of synaptophysin suggests that the protein may function as a channel in the synaptic vesicle membrane, with the carboxyl terminus serving as a binding site for cellular factors.

Amino Acid Sequence↗