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Ribosomal RNA in Mauthner axon: implications for a protein synthesizing machinery in the myelinated axon.

RNA was extracted from myelin-free Mauthner axons of the goldfish on a microscale and fractionated by microelectrophoresis. Microextracts showed the presence of nominal 26 SE, 18 SE, 5 SE and 4 SE components, which co-migrated with rRNA from fish brain. In addition, a non-ribosomal 15 SE component was present in axon microextracts, but not in RNA extracts of fish brain or of myelin sheath from Mauthner axon, indicating an unusual enrichment of a putative mRNA class. Evidence was presented to support the contention that axonal rRNA was not due to contamination from the myelin sheath. Possible reasons for the lack of ultrastructural evidence for axoplasmic ribosomes are discussed.

Animals

"Giant axonal neuropathy" caused by industrial chemicals: neurofilamentous axonal masses in man.

Symmetrical polyneuropathy developed in two patients after they had been in contact with acrylamide and methyl n-butyl ketone, respectively. In sural nerve biopsy material from both patients, electron microscopy showed frequent focal axonal swellings containing masses of neurofilaments. Some axons undergoing axonal degeneration also were seen. These morphologic features are identical to those produced in experimental animals after exposure to these chemicals and are similar to those found in n-hexane neuropathy and in the three reported cases of giant axonal neuropathy. Sural nerve biopsy is an important diagnostic test in identifying cases of peripheral neuropathy caused by these chemicals.

Acrylamides

A physical model of nerve axon. II: Action potential and excitation currents. Voltage-clamp studies of chemical driving forces of Na+ and K+ in squid giant axon.

An adsorption model of nerve axon has been extended to account for the origin of membrane currents observed under voltage-clamp conditions. Differing from the Hodgkin-Huxley model, which attributes excitation solely to a change of ionic conductances of the membrane, the present model proposes that a layer of axoplasm attached to the membrane (axon cortex) can undergo conformational changes and hence modulate selectivity for mobile ions. To test the model, a two-step voltage-clamp study was made of the chemical driving forces of Na+ and K+ ions in squid giant axon. The forces were measured by determining the instantaneous current-voltage relation when membrane current is carried by Na+ only or K+ only. The data indicate that the chemical driving force varies as a function of time and does not agree with the Nernst relation during the early phase of excitation. Implications of the observations are discussed.

Action Potentials

Slow axonal transport or proteins; blockade by interruption of contact between cell body and axon.

The influence of ligation and colchicine treatment on the axonal transport of slowly migrating [3H]leucine-labelled proteins was studied in the vagus nerve of the rabbit. Two days after [3H]leucine labelling of the dorsal motor nucleus of the vagus nerve, ligation or local application of 60 mM colchicine immediately blocked the further progression of slowly migrating proteins distal to the site of treatment. Application of 50-100 mug colchicine to the nerve cell bodies 2 days after labelling blocked the transport of slowly migrating proteins within the next 24 h. It is suggested that contact between nerve cell body and the axon is necessary for the maintenance of the slow transport of proteins in these nerves.

Animals

Axonal transport and axonal processing of low molecular weight proteins from the abdominal ganglion of Aplysia.

Axonal transport of proteins in nerves of the abdominal ganglion of Aplysia was observed after a 2 h incubation of the ganglion in tritiated amino acids. The transported proteins migrate as a series of discrete peaks, all apparently moving at a rate of 3 mm/h. This process is sensitive to both colchicine and vinblastine, the former agent reducing the amount of transported material without affecting the transport rate. The molecular weight distribution of the transported proteins, as revealed by polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate (SDS), is basically unchanged for up to 20 h after labeling. Low molecular weight species (less than or equal to 18,000 daltons) make up 10-20% of the transported protein and appear to be enriched in leucine. These proteins undergo proteolytic cleavage during transport, eventually reaching a molecular weight of 3000 daltons or lower. It is suggested that these data reflect the axonal transport and processing of neurosecretory peptides synthesized by identifiable neurons of the ganglion.

Animals

Tracing axons and axon collaterals of spinal neurons using intracellular injection of horseradish peroxidase.

Intracellular injection and subsequent histochemical localization of horseradish peroxidase have been used to stain the soma, dendrites, axons, and axon collaterals of spinalcervical tract neurons and unidentified dorsal horn neurons in the cat. This technique may be used in combination with the intracellular injection of Procion yellow to demonstrate by light microscopy connections between physiologically typed vertebrate neurons.

Animals

The intra-axonal transport of acetylcholine and cholinergic enzymes in rat sciatic nerve during regeneration after various types of axonal trauma.

The proximo-distal intra-axonal transport of acetylcholine (ACh) and cholinergic enzymes (choline acetyltransferase, CAT, and ACh-esterase, AChE) in rat regenerating sciatic nerve was studied by accumulation technique. Four types of axonal trauma were performed: freezing with solid CO2, crushing, ligating the nerve with remaining tight silk ligature, and cutting the nerve. Normal and sham-operated rats were used as controls. One to twenty-nine days later, the nerves were crushed about 15 mm proximal to the trauma. The nerve segment proximal to this crush was dissected out 12 hr later and assayed for ACh-content and enzyme activities. The increase in this segment 12 hr after crushing was taken as an indication of proximo-distal transport in the regenerating nerves. ACh transport did not seem to vary during regeneration as compared to controls. In contrast, the transport of both CAT and AChE was initially markedly depressed. Towards the end of the observation period (29 days), a recovery of CAT-transport occurred in all groups. Recovery of AChE-transport was marked in the freeze and crush groups. In the cut group no recovery was seen and in the ligated group only a small recovery occurred. Thus, in the nerves where regeneration was facilitated by the presence of intact connective tissue sheaths (freezing and crushing) recovery of transport occurred earlier than in cut or ligated nerves.

Acetylcholine

Taurine in the developing rabbit visual system: changes in concentration and axonal transport including a comparison with axonally transported proteins.

[35S]Taurine injected intravitreally into rabbits was transported axonally to the optic nerve terminals. Considerably more [35S]taurine was transported in young rabbits than in mature rabbits. The time course of taurine transport did not parallel that of proteins labeled with [3H]proline in the same system. The concentration of taurine in all components of the visual system, except retina, was greater in young animals than in mature animals, and was especially high in optic nerve. The possible functions of the high concentrations of taurine and the greater amount of axonally transported taurine in developing mammalian CNS are discussed.

Age Factors

Localization of horseradish peroxidase-alpha-bungarotoxin binding in crustacean axonal membrane vesicles and intact axons.

A conjugate of alpha-bungarotoxin with horseradish peroxidase was used to visualize alpha-bungarotoxin binding sites at the fine structural level in isolated axonal membrane vesicles from lobster walking leg nerve. These plasma membrane vesicles have previously been shown to exhibit saturable binding of [3H]nicotine and [3H]acetylcholine. Binding of the toxin was identified in the axon plasma membrane and could be blocked by pretreatment with excess free alpha-bungaratoxin or d-tubocurarine. Binding sites for alpha-bungarotoxin were identified by the same technique in sections of intact nerve fibers from both lobster and spider crab and were found to be localized primarily in the axolemma rather than in the Schwann cell membrane.

Acetylcholine

Cinematographic analysis of contractile events produced in intrafusal muscle fibres by stimulation of static and dynamic fusimotor axons.

1. Muscle spindles with an intact blood supply and uninterrupted connexions with ventral and dorsal spinal roots (Bessou & Pagés, 1967, 1972) have been prepared in cat's tenuissimus muscles with the aim of cinephotographically recording intrafusal movements induced by the stimulation of single static or dynamic gamma axons; the time cours of these movements and the morphological kind of activated intrafusal muscle fibres have been established. 2. Displacements of spindle guiding marks in the equatorial region elicited by stimulating single static gamma axons are 4-20 times greater in amplitude than the ones elicited by stimulating dynamic gamma axons at the same frequency. 3. The dynamic gamma axons induced a contraction only in nuclear bag fibres which, in addition, never received any static gamma innervation. The static gamma axons evoked contractions either in nuclear bag fibres alone, or in nuclear chain fibres alone, or in both types of intrafusal fibres. Two thirds of static gamma axons supplied nuclear bag fibres. For various reasons, one half only of static gamma axons innervating nuclear bag fibres could be shown to simultaneously innervate nuclear chain fibres. Consequently, about one third of static gamma axons supplied both nuclear bag fibres and nuclear chain fibres, but it is highly probable that this latter figure is an underestimate. One third of static gamma axons produced contraction in nuclear chain fibres only. In this work, the distribution of fusimotor axons has been established in only one muscle spindle of the cluster of muscle spindles that each fusimotor axon is generally innervating. 4. Generally speaking, a static gamma axon elicits contraction of several intrafusal fibres whereas a dynamic gamma axon innervates only one intrafusal fibre and frequently only one pole of the fibre. 5. One third of static gamma axons evoked contractions in nuclear chain fibres that seemed to involve the whole pole. The other static gamma axons and all dynamic gamma axons produced, in the intrafusal fibres that they supplied, one or several foci of localized contractions. 6. The nuclear chain fibres contract and relax faster than nuclear bag fibres. The contractions of nuclear bag fibres supplied by static gamma axons are stronger and faster than those of nuclear bag fibres innervated by dynamic gamma axons. Nearly all nuclear bag fibres innervated by static gamma axons, like the nuclear chain fibres, show transient contractions at each pulse of a stimulation at low frequency (2-20/sec). 7. The results are discussed taking into account the available anatomical and physiological data on the muscle spindle. Their consequences with regard to intrafusal working are briefly considered.

Animals