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B Droz

Publications and source records attributed to B Droz.

At least 55 records · Page 3Linked to original sources

Contribution of axonal transport to the renewal of myelin phospholipids in peripheral nerves. I. Quantitative radioautographic study.

Kinetics of phospholipid constituents transferred from the axon to the myelin sheath were studied in the oculomotor nerve (OMN) and the ciliary ganglion (CG) of chicken. Axons of the OMN were loaded with transported phospholipids after an intracerebral injection of [2-3H]glycerol or [3H]labeled choline. Quantitative electron microscope radioautography revealed that labeled lipids were transported in the axons mainly associated with the smooth endoplasmic reticulum. Simultaneously, the labeling of the myelin sheath was found in the Schmidt-Lanterman clefts and the inner myelin layers. The outer Schwann cell cytoplasm and the outer myelin layers contained some label with [methyl-3H]choline, but virtually none with [2-3H]glycerol. With time the radioactive lipids were redistributed throughout and along the whole myelin sheath. Since [2-3H]glycerol incorporated into phospholipids is practically not re-utilized, the occurrence of label in myelin results from a translocation of entire phospholipid molecules and from their preferential insertion into Schmidt-Lanterman clefts. In this way, the axon-myelin transfer of phospholipid contributes rapidly to the renewal of a limited pool of phospholipids in the inner myelin layers. When [methyl-3H]choline was used as precursor of phospholipids, the rapid appearance of the label in the inner myelin layers was interpreted also as an axon-myelin transfer of labeled phospholipids. However, the additional labeling of the outer Schwann cell cytoplasm adjacent to Schmidt-Lanterman clefts and of the outer myelin layers reflects a local re-incorporation of the base released from the axon. By these two processes, the axon contributes to purvey the inner myelin layers with new phospholipids and the Schwann cells with new choline molecules.

Animals↗

Contribution of axonal transport to the renewal of myelin phospholipids in peripheral nerves. II. Biochemical study.

The classes of radioactive phospholipids appearing in the ciliary ganglion (CG) and especially in the myelin sheath of the intraorbital part of the oculomotor nerve (OMN) were determined after the intracerebral injection of [2-3H]glycerol and [methyl-14C]choline to chickens. Analysis of the radioactive compounds in water-soluble fractions and chloroform-methanol extracts was performed by thin-layer chromatography (TLC). The water-soluble content of the OMN and CG was much poorer in [2-3H]glycerol and metabolites than in [methyl-14C]choline and derivatives. All classes of glycerophospholipids were found to be axonally transported along the OMN and into the CG, but choline-phosphoglycerides (CPG) were largely predominant. In myelin fractions from the OMN, the specific radioactivity (SRA) of CPG labeled with [2-3H]glycerol reached a maximum earlier (40 h) than the SRA of CPG labeled with [methyl-14C]choline. A 25-fold enhancement of the [14C]SRA of sphingomyelin (SM) was observed between 12 h and 7 days. These results indicate that: (1) axonally transported phospholipids labeled with [2-3H]glycerol consist mainly of CPG; (2) small amounts of CPG are translocated from the axon to myelin; and (3) the progressive enrichment of myelin in [14C]CPG and, to a greater extent, SM draws attention to the importance of the base recycling for local synthesis of myelin phospholipids. Thus the axonal supply of Schwann cells with choline and the transfer of axonal phospholipids to myelin would probably contribute to the metabolic interdependence existing between neuron and glia.

Animals↗

'Acrylamide-induced' neuropathy and impairment of axonal transport of proteins. I. Multifocal retention of fast transported proteins at the periphery of axons as revealed by light microscope radioautography.

The axonal transport of proteins was studied in ciliary ganglia of control and acrylamide-treated chickens. After an intracerebral injection of [3H]lysine, the distribution of axonally transported proteins was analyzed in the preterminal segments of preganglionic axons and in their caliciform nerve endings by means of quantitative light microscope radioautography. By 7 days after the injection of [3H]lysine, the concentration of labeled proteins in the preganglionic axons was either similar or slightly increased in acrylamide-treated chickens as compared to controls. In contrast, by 3 h, whereas the axons of controls were poorly radioactive, 20-30% of the axons in acrylamide-treated chickens displayed focal and intense labeling at their periphery; simultaneously, in acrylamide treated chickens, 20-30% of the nerve endings contained a decreased amount of radioactive proteins. It is concluded that acrylamide induces multifocal retention of fast axonally transported proteins in preterminal segments of certain axons.

Acrylamides↗

'Acrylamide-induced' neuropathy and impairment of axonal transport of proteins. II. Abnormal accumulations of smooth endoplasmic reticulum as sites of focal retention of fast transported proteins. Electron microscope radioautographic study.

The distribution of fast axonally transported proteins was studied by electron microscope radioautography in ciliary ganglia of chickens treated or not treated with acrylamide. At 3 h after the intracerebral injection of [3H]lysine, the preganglionic axons of the untreated chickens displayed few silver grains, mainly associated with smooth endoplasmic reticulum (SER) profiles. In most axons of acrylamide-treated chickens, a similar pattern was observed, except in axons which exhibited focal and intense labeling underneath the axolemma: clusters of silver grains indeed overlayed peripheral accumulations of tubulovesicular profiles of SER, dense core vesicles and mitochondria. After impregnation with heavy metals, electron microscope observation of 1 micrometer thick sections showed a locally disorganized SER forming a complex network of tubules intermingled with vesicles and mitochondria. Such a local disorganization of the peripheral SER in the distal part of the axons, could be responsible for the focal stasis of fast transported proteins; it seems to be one of the earliest changes detectable in axons damaged by acrylamide treatment.

Acrylamides↗

Calcium metabolism and acetylcholine release at the nerve-electroplaque junction.

1. Calcium metabolism was investigated in the electric organ of Torpedo at rest and during synaptic activity. When the tissue was incubated in high calcium concentrations, complete exchange was obtained between cellular calcium and that of the external medium. In the presence of low concentrations, the tissue retained its calcium which, in this case, was poorly exchanged. 2. Stimulation of the nerves to the electric organ provoked a net increase in cellular calcium and an acceleration of its exchange with that of the extracellular space. This entry of calcium seemed to involve mainly the presynaptic axon endings since it occurred even when transmission was blocked by curare. 3. The presynaptic accumulation of calcium was accompanied by a decrease in the vesicular pool of acetylcholine (ACh). On the other hand, the level of extracellular Ca2+ seemed to be critical for the intraterminal repartition of ACh between the vesicular pool and the cytoplasmic pool, and also for exchange of high energy phosphate between ATP and thiamine esters. 4. Radioautography of 45Ca at the electron microscope level confirmed that most of the Ca accumulation following stimulation concerned the presynaptic axon endings. The accumulated 45Ca was closely associated with the plasma-lemma region of nerve terminals, mainly in the region of active zones. 5. Taking this into account, and also the fact that synaptic vesicles have a very efficient Ca sequestration mechanism (see Israël et al., this symposium) it is proposed that, after entry, Ca is taken up by the vesicles situated in active zones, exchanged against vesicular ACh, concentrated in the vesicle and finally released by exocytosis.

Acetylcholine↗

[Visualization of opiate receptors in the locus coeruleus of rats: high resolution radioautography after administration of a tritiated met-enkephalin analog].

Opiate receptor sites were visualized by high resolution radioautography in the locus coeruleus of the Rat following intraventricular injection of the met-enkephalin analogue FK 33-824 tritiated with a high specific activity (3H-FK). After administration of a tracer dose of 3H-FK, more than 75% of the radioactivity detected in the locus coeruleus is specifically bound to opiate receptor sites. These are distributed both between and inside neuronal perikarya. After administration of 3H-FK at high concentration, electron microscope radioautography shows the presence of specific opiate binding sites at the level of axo-somatic and axo-dendritic synaptic junctions. These synaptic binding sites could correspond to receptor sites for endogenous enkephalins.

Animals↗

The smooth endoplasmic reticulum: structure and role in the renewal of axonal membrane and synaptic vesicles by fast axonal transport.

The spatial arrangement of the smooth endoplasmic reticulum (SER) was studied in 0.5-2 mum thick sections of rat spinal and chick ciliary ganglia previously impregnated with heavy metal salts. Electron microscopy at low (10-5 V) or high (10-6 V) voltage showed the impregnated SER as a continuous system extending probably from the perikaryon to the axon terminal. Tubules of the SER, which were running in a parallel direction with the axon, were occasionally seen in close apposition with the axonal membrane. Moreover in the preterminal region, anastomosed tubules of the SER formed a subsurface 'primary network' and gave rise to a deeper 'secondary network' made of thinner tubules; synaptic vesicles bulging at the tip of thin tubules of the SER were frequently observed. To specify the role played by the SER in the fast axonal transport, chicken ciliary ganglia were slighty compressed and radioautographed 3 h after the intracerebral injection of [3-H]lysine. Quantitative analysis of the silver grain distribution indicated that labeled proteins, rapidly conveyed down the axon, piled up in regions containing an accumulation of SER profiles. On the basis of these results, it is concluded that: (1) the SER appears as a continuous intraaxonal pathway bridging the perikaryon and the axon terminal; (2) the SER conveys macromolecular components with the fast axonal transport; (3) the conveyed macromolecules, which are delivered to the axonal membrane and to the synaptic vesicles, are probably transferred by means of connections with the SER.

Animals↗