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Prevention of defective axonal transport in streptozocin-diabetic rats by treatment with "Statil" (ICI 128436), an aldose reductase inhibitor.

This investigation examined the effect of treatment of streptozocin-diabetic rats with the aldose reductase inhibitor "Statil" (25 mg/kg/day p.o.) on axonal transport in cholinergic neurons of the sciatic and vagal nerves and on nerve polyol and sugar levels. Three weeks of experimental diabetes caused deficits in the accumulation of choline acetyltransferase activity proximal to 24-h constrictions in the left sciatic and vagus nerves. These deficits did not develop in age-matched, similarly diabetic rats that were treated with the aldose reductase inhibitor. The inhibitor prevented completely the build-up of sorbitol and markedly reduced the build-up of fructose in the sciatic nerves of the treated diabetic rats. The inhibitor also prevented the depletion of myo-inositol that was seen in the untreated diabetic animals. It is suggested that these findings indicate a possible approach to the elucidation of the pathogenesis of cardiac vagal dysfunction in diabetes mellitus.

Animals↗

Pulpal exposure alters neuropeptide levels in inflamed dental pulp and trigeminal ganglia: evaluation of axonal transport.

Dental pulp is richly innervated with neurons containing calcitonin gene-related peptide (CGRP) or substance P (SP). Prior studies have demonstrated that inflammation alters these pulpal neuropeptides. In this study, we used a radioimmunoassay to evaluate the specificity of this response and the contribution of axonal transport. Rat mandibular molars were exposed and immunoreactive CGRP (iCGRP) and immunoreactive SP were measured. At 7 to 14 days after exposure, both pulpal iCGRP (73%) and immunoreactive SP (135%) displayed peak increases above control levels. This response was somatotopically restricted, with no changes observed in contralateral (intact) molars, or in ipsilateral mandibular molars after exposure of maxillary molars. Transection of the inferior alveolar nerve on day 13 significantly reduced pulpal levels of iCGRP on day 14. Collectively, these studies indicate that pulpal inflammation evokes a selective alteration in neuropeptide levels, due at least in part to alterations in transport or synthesis of neuropeptides in the trigeminal ganglion.

Analysis of Variance↗

Microtubules provide directional cues for polarized axonal transport through interaction with kinesin motor head.

Post-Golgi carriers of various newly synthesized axonal membrane proteins, which possess kinesin (KIF5)-driven highly processive motility, were transported from the TGN directly to axons. We found that KIF5 has a preference to the microtubules in the initial segment of axon. Low dose paclitaxel treatment caused missorting of KIF5, as well as axonal membrane proteins to the tips of dendrites. Microtubules in the initial segment of axons showed a remarkably high affinity to EB1-YFP, which was known to bind the tips of growing microtubules. These findings revealed unique features of the microtubule cytoskeletons in the initial segment, and suggested that they provide directional information for polarized axonal transport.

Animals↗

The mechanism and regulation of fast axonal transport.

Recent in vitro studies of microtubule-dependent organelle movement have provided a great deal of information on the molecular mechanism of fast axonal transport. Microtubule-dependent organelle movement occurs in most cells, but in neurons active transport is absolutely necessary for materials to travel from the cell body to the synapse. Since fast transport is crucial for neuronal survival, it is likely that specialized regulatory mechanisms have been developed. It is clear that the microtubule-based motors, kinesin and cytoplasmic dynein are the enzymes that power organelle motility; however, additional cytoplasmic components are required to create an 'organelle translocation complex' that is competent for transport. Organelle transport might be regulated at the level of any of these components, i.e. the motors, their accessory factors, or the organelle binding sites. The direction of organelle movement is probably governed by the membrane binding site. In this review we discuss these topics and consider the mechanism of transport of the retrograde motor, cytoplasmic dynein, to the nerve terminal, and possible ways that unidirectional transport could occur on the non-polarized array of microtubules found in some dendrites.

Adenosine Triphosphatases↗

Labelling by axonal transport of myelin-associated proteins in the rabbit visual pathway.

After intraocular injections of [3H]leucine, six regions of the visual pathway of adult rabbit were used to study the spatio-temporal pattern of the slow anterograde axonal transport of radioactive proteins associated with the particulate fraction, the water-soluble fraction and the myelin fraction. Unlike other fractions, myelin-associated labelled proteins represented a time-constant (for a given region) percentage of total tissue radioactivity. This percentage increased from the first half to the second half of the optic nerve and remained high in the chiasma and tract. The peak specific radioactivity of myelin decreased in the same direction. Myelin proteins were separated by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and the labelling patterns obtained in different regions and at different survival times were compared. At the peak of myelin radioactivity of a given region the label was typically associated with four protein bands, L1, L2, L3 and L4, of 40000, 44000, 62000, and 68000 mol.wts. respectively. The basic protein, the proteolipid protein and the W1 component (mol.wt. 51000-53000) of the Wolfgram proteins were not significantly labelled. The radioactivity associated with the W2 component (mol.wt 60000) of the Wolfgram proteins could be derived from the closely migrating L3 component. At shorter survival times no clear labelling pattern could be detected. At longer survival times radioactivity was almost totally localized around band L3. The results presented underline the importance of choosing appropriate experimental conditions to obtain a consistent labelling pattern of myelin-associated proteins and to investigate the possible mechanism responsible for this phenomenon.

Animals↗

Electron microscopic study of the orthograde axonal transport of horseradish peroxidase in the supraoptico-neurohypophysial tract of the rat.

Horseradish peroxidase (HRP) has been used as a protein tracer in order to visualize the ultrastructural sites of the orthograde transport of protein macromolecules in the hypothalamo-neurohypophysial tract of the rat. After a local injection of HRP within the supraoptic nucleus, the reaction product was observed: (1) mainly in tubules of the smooth endoplasmic reticulum in the more proximal part of the axons, and (2) in granules and microvesicles of the axon terminals. Observations on thick sections clearly showed the existence of a relationship between the smooth endoplasmic reticulum containing HRP and the labeled granules or microvesicles. These data are in good agreement with previous findings showing the existence of direct continuity between tubules of the smooth endoplasmic reticulum and a fraction of the neurosecretory granules and microvesicles. This evidence further reinforces the hypothesis that the latter organelles may possibly originate locally in the axons from the tubules of the smooth endoplasmic reticulum which may therefore be proposed as a possible vehicle for a non-granular intra-axonal transport of neurosecretory material in neurosecretory neurons.

Animals↗

Lysosomal activity in developing cat alpha-motor axons under normal conditions and during retrograde axonal transport of horseradish peroxidase.

The occurrence of acid phosphatase (AcPase)-positive bodies, i.e., lysosomes, in lumbosacral alpha-motor axons of kittens, 0-16 weeks of age, was analyzed by light and electron cytochemical methods under normal conditions and after intramuscular injection of horseradish peroxidase (HRP). Axonal lysosomes were rare early postnatally. In 3-week-old animals, a few AcPase-positive bodies appeared in the axoplasm at some nodes of Ranvier in the peripheral nervous system (PNS) and internodally in the intrafunicular motor axon parts within the central nervous system (CNS). From 6 weeks postnatally, a nodal concentration of AcPase-positive bodies was also noted in the CNS. The number of AcPase-positive bodies continued to increase gradually in the course of neuronal maturation. In 16-week-old animals, axonal AcPase activity was still at considerably lower levels than at adult stages. At all ages, acid hydrolase-containing organelles were most commonly found at ventral root nodes. After injection of HRP in the medial gastrocnemius muscle, accumulations of AcPase-positive bodies were seen in the axoplasm at some PNS nodes of the HRP-injected sides of kittens aged 8, 12, and 16 weeks. Incubation for demonstration of both HRP and AcPase activity showed that some organelles at HRP-transporting nodes contained both types of reaction product. The nodal AcPase activity in the intrafunicular, CNS parts of alpha-motor axons of the HRP-exposed sides did not differ from that of the contralateral, uninjected sides. In view of our previous observations in alpha-motor neurons of adult cats in which a lysosome-mediated degradation of axonally transported materials may take place at PNS nodes of Ranvier, the present study illuminates possible differences in the ability to interfere with axonal transport between developing and mature neurons. The infrequent presence of lysosomes in developing alpha-motor axons and the implied disability of their nodal regions to interfere with axonally transported constituents in a way similar to that seen in adult animals may be of significance in that trophic and chemical signals can pass unhindered between the periphery and perikaryon. However, this could also have negative consequences for the vulnerable immature neuron in that various materials retrieved at the axon terminals outside the CNS are permitted a more-or-less free access to the perikaryon.

Aging↗

Retrograde axonal transport of asialoglycoproteins in mouse trigeminal neurons in vivo and in rat dorsal root ganglia neurons in vitro.

Different terminal sugars of the glycoprotein orosomucoid were exposed by sequential glycosidase digestions. The orosomucoid and its different derivatives were conjugated to horseradish peroxidase by a two-step glutaraldehyde coupling procedure, injected into the snout of 12-day-old mice or exposed to dorsal root ganglia neurons from embryonic rats, cultivated in a two chamber system. A marked increase in transport of the conjugates in the trigeminal and dorsal root ganglia neurons was observed histochemically after removal of sialic acid, exposing galactose as the terminal sugar. Quantitative hydrolysis of galactose residues resulted in reduced uptake. The data suggest the presence of a galactose-recognition molecule in the axon-terminal membrane, involved in retrograde axonal transport.

Animals↗

Effect of 2,5-hexanedione and 3,4-dimethyl-2,5-hexanedione on retrograde axonal transport in sciatic nerve.

The effects of systemically introduced neurotoxic solvents 2,5-hexanedione (2,5-HD) and 3,4-dimethyl-2,5-hexanedione (DMHD) on retrograde axonal transport (RT) of 125I-labeled tetanus toxin (TT) was studied in rat and mouse sciatic nerves. The rate of retrograde transport of TT in control rat sciatic nerves was slightly higher (6.8 +/- 0.4 mm/h) than in mouse sciatic nerves (5.4 +/- 0.5 mm/h). A single high dose of 2,5-HD (1,000 mg/kg, i.p.) produced a time-dependent effect on RT in mouse sciatic nerves. 2,5-HD caused a gradual decrease in the velocity of RT (approximately 65% inhibition between 2.0-2.5 h) with a reversal to normal rate 3-5 h after the toxin administration. The effect of DMHD on RT was examined following semi-chronic treatment in rats. DMHD caused a significant decrease (approximately 50%) in the rate of TT transport, in addition, it produced weight loss and hind-limb paralysis.

Animals↗

Drosophila Lis1 is required for neuroblast proliferation, dendritic elaboration and axonal transport.

Haplo-insufficiency of human Lis1 causes lissencephaly. Reduced Lis1 activity in both humans and mice results in a neuronal migration defect. Here we show that Drosophila Lis1 is highly expressed in the nervous system. Lis1 is essential for neuroblast proliferation and axonal transport, as shown by a mosaic analysis using a Lis1 null mutation. Moreover, it is cell-autonomously required for dendritic growth, branching and maturation. Analogous mosaic analysis shows that neurons containing a mutated cytoplasmic-dynein heavy chain (Dhc64C) exhibit phenotypes similar to Lis1 mutants. These results implicate Lis1 as a regulator of the microtubule cytoskeleton and show that it is important for diverse physiological functions in the nervous system.

Animals↗

Retrograde axonal transport of true blue dye by the peripheral autonomic nerves in canine nasal mucosa.

In this study we employed retrograde axonal transport of (E)-2,2'-vinylendi-benzofuran-5-carboxamidin-diaceturate+ ++ or true blue (TB) to study the peripheral autonomic innervation of the canine nasal mucosa. After injection of TB into the nasal mucosa, labeled neurons were found in the ipsilateral sphenopalatine ganglion (SPG) and the superior cervical ganglion (SCG). There were no labeled neurons in the middle cervical or stellate ganglia. This indicated that the origin of the postganglionic sympathetic fibers of the nasal mucosa was only from the ipsilateral SCG. When TB was injected into the nasal mucosa of dogs following a caudal or ethmoidal neurectomy, labeled neurons could still be found in the SPG and SCG. When TB was injected into the nasal mucosa of dogs following ethmoidal and vidian neurectomies or with maxillary neurectomy added, some labeled neurons could still be found in both the ipsilateral SPG and SCG. These results support the concept that another pathway--perhaps perivascular--exists for postganglionic sympathetic fibers other than the vidian and ethmoidal nerves. Labeled neurons were still observed in SPG when TB was applied to the canine nasal mucosa following neurectomy of either the ethmoidal or the caudal nasal nerve. However, retrograde labeled neurons could not be found in SPG following simultaneous neurectomies of the ethmoidal and caudal nasal nerves. These results show that the postganglionic parasympathetic fibers originating in the SPG travel along the ethmoidal and caudal nasal nerves.

Animals↗

Anterograde axonal transport of [125I]wheat germ agglutinin by motor neurons innervating the rat lateral rectus muscle.

Despite widespread evidence of the affinity of wheat germ agglutinin (WGA) for sensory, preganglionic autonomic and other CNS neuron cell bodies, evidence of the uptake and anterograde transport of WGA by motor neurons innervating skeletal muscle is lacking. It was hypothesized that these cells might lack plasma membrane moieties with N-acetylglucosamine for selective uptake and transport of WGA. To test this, rats were anesthetized and injected intracisternally with affinity-purified [125I]-WGA in order to label abducens motor neurons. Based on results of light and electron microscopic autoradiography, it can be concluded that abducens motoneurons, and presumably other motoneurons, are capable of uptake and anterograde axonal transport of [125I]WGA.

Abducens Nerve↗