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PubMed1981

Retrograde axonal transport mediates the onset of regenerative changes in the hypoglossal nucleus.

The hypoglossal nucleus was assayed for [14C]2-deoxyglucose uptake 24 h after axotomy with and without colchicine injections into the nerve proximal to the nerve transection. Colchicine effectively blocked the usual increase in [14C]2-deoxyglucose uptake seen after axotomy. The drug also blocked the transport of horseradish peroxidase from the tongue muscles to the hypoglossal nucleus which otherwise occurred within this time period. Distribution of [3H]colchicine showed that the drug remained localized within the nerve close to the injection point. These results suggest that retrograde axoplasmic flow is involved in the mechanism which initiates hypoglossal neuron regeneration.

Animals↗
PubMed1977

Axonal transport of labeled material into sensory nerve ending of cat carotid body.

The origin of nerve endings on glomus cells in the carotid body has been the subject of much controversy in recent years. Specifically, the problem is whether these nerve endings, which contain clear-core ("synaptic") vesicles and mitochondria arise from sensory neurons in the petrosal ganglion or from efferent neurons located in the brain stem or elsewhere. To study this problem, [3H]proline was applied to cat petrosal ganglia, the animals were allowed to survive for 3 h-7 days, and the peripheral distribution of the label was analyzed by sample oxidation/scintillation counting and by EM autoradiography. The time courses of distribution of label along the nerves and the accumulation of label in the carotid body indicated the presence of fast, intermediate and slow components of axonal flow. EM autoradiographs of carotid bodies showed the label localized almost exclusively to nerve fibers and to nerve terminals on glomus cells. As much as 60--90% of the nerve terminals were labeled in a given ultrathin-section autoradiograph. Passive movement of label from the injection site, or fibers of passage (efferent) through the ganglion, did not contribute to the labeled material since administration of [3H]proline onto the desheathed nerve away from the ganglion was ineffective in labeling the carotid body. The results suggest that most, if not all, nerve terminals on glomus cells in cat carotid body arise from neurons in the petrosal ganglion.

Animals↗
PubMed1984

Axonal transport of opiate receptors in capsaicin-sensitive neurones.

Opiate receptors measured in vitro or in vivo with [3H]lofentanil in the rat vagus nerve were found to accumulate on both sides of a ligature, thus indicating a bidirectional axoplasmic transport of these receptors. When rats were treated with capsaicin, the accumulation of opiate receptors was tremendously reduced in the vagus whereas muscarinic receptors in ligated sciatic nerves were unaffected. Since capsaicin is known to affect sensory neurones, mostly those containing substance P, the present results support the idea that the opiate receptors in the vagus are associated with substance P neurones.

Animals↗
PubMed1975

Association of proteins undergoing slow axonal transport with goldfish visual system myelin.

[3H]Proline derived radioactivity which is slowly transported in the goldfish optic nerve has been isolated in association with purified preparations of optic tectal myelin. Acrylamide gel analysis reveals that most of the slowly transported radioactivity is distributed among the high molecular weight myelin proteins with a major peak comigrating with a band (approximately 42,000 daltons) which is prominent in goldfish brain myelin. A similar peak is not seen in gels of whole synaptosome protein. Relatively small but measureable amounts of radioactivity comigrate with the basic and proteolipid proteins. A low density myelin subfraction is 3-5 fold lower in specific radioactivity (disint./min/mug protein) than a corresponding high density fraction and shows particularly low specific radioactivities in the basic and proteolipid bands. The results are discussed with reference to the possible adherence of specific axonal membrane fragments as well as other potential sources of myelin label.

Animals↗
PubMed1989

A long term study of axonal transport in the central visual system following eye enucleation in the adult cat.

The effect of the enucleation of one eye on anterograde and retrograde labelling in geniculo-cortical, cortico-geniculate and commissural projections was investigated in adult cats by means of horseradish peroxidase (HRP) and tritiated aminoacids. It was found that in addition to the immediate decrease of retrograde labelling with HRP in the cortical projections from the deafferented A-laminae of the dorsal part of the lateral geniculate nucleus (Singer et al. 1977) there is a further reduction which lasts up to 75 days after enucleation. At 146 and 363 days after enucleation a slight increase in the number of labelled neurones was noted in the deafferented lamina. Qualitative assessment did not reveal any changes of anterograde labelling with tritiated amino acids in geniculo-cortical, cortico-geniculate and commissural axones. In addition, the retrograde labelling with HRP in cortico-geniculate and commissural projections seemed to be unaffected by eye enucleation.

Amino Acids↗
PubMed2003

Axonal transport of British and Danish amyloid peptides via secretory vesicles.

The ABri and ADan amyloid peptides deposited in familial British and Danish neurodegenerative disorders are generated by processing mutant forms of the precursor protein BRI2. Although the pathogenic process that leads to deposition of amyloid in the brains of patients has been studied extensively, the cellular processes and normal function of the precursor protein did not receive much attention. We observed in a variety of transfected cell lines the presence of two independent proteolytic processing events. In addition to the previously described cleavage, which results in the production of carboxyl-terminal approximately 3 kDa wild-type peptide or approximately 4 kDa ABri or ADan peptides, we describe a novel amino-terminal cleavage site within BRI2. Both cleavages occur within the cis- or medial-Golgi. Following cleavage, the BRI2-derived carboxyl-terminal peptides are transported via a regulated secretory pathway into secretory vesicles in neuronal cells. Worth noting is that expression of wild-type British or Danish mutants of BRI2, in mouse neuroblastoma N2a cells that do not express endogenous BRI2, induces elongation of neurites, which suggests a role for this protein in differentiation of neuronal cells.

Adaptor Proteins, Signal Transducing↗
PubMed1981

[Evolution of the storage site of iron-dextran after retrograde axonal transport in the rat central nervous system (author's transl)].

The uptake, transport, and storage of an iron-dextran complex (used as a neuroanatomical tracer in the nigrostriatal pathway of rat) by neurons of the CNS is described. The complex is taken up by endocytosis and transported by lysosomes. Some iron-containing mitochondria are observed within the nigral cell bodies. The neuronal labelling remains for 10 days. Thereafter, the iron labelling of neurons decreases. No important neuronal degeneration is observed as a result of iron toxicity. The iron lost by neurons is taken up by glial cells. It is converted into ferritin and increases the intracerebral endogenous iron content.

Animals↗
PubMed1987

Changes in axonally transported proteins in the mature and developing rat nervous system during early stages of methyl mercury exposure.

It was established by means of SDS polyacrylamide gel electrophoresis that direct injections of methyl mercury (10 micrograms Hg) into the mature rat vitreous body of the eye decrease protein synthesis in the retina and optic nerve at 4 hours after injection. Although the global spectrum of polypeptides did not change, a specific decrease in the volume of polypeptides of 20-23 K daltons molecular weight was evident. Conversely, systemic exposure to methyl mercury resulted in increased protein synthesis of polypeptides of 20-23 K molecular weight both in adult (8 mgHg/kg/day for 8 days) and neonatal rats (2 mgHg/kg/day for 10 days). In addition, specific changes in the volume of polypeptides 75-90 K molecular weight were noted in sciatic nerves of neonatal rats. These data are consistent with a bimodal response in protein synthesis following MeHg treatment. Local presence of MeHg following direct injection into the eye causes a reduction in protein synthesis, while chronic systemic exposure results in increased synthesis and transport of proteins in both mature and developing optic nerves and neonatal sciatic nerves. Thus, these systems possess the capacity to attempt regenerative processes through induction of a small subset of proteins known as GAPs (Growth-Associated Proteins) during the early stages of systemic methyl mercury exposure. These wide spread and system-specific changes are consistent with growth-specific functions during the early stages of methyl mercury exposure.

Aging↗
PubMed1987

Calmodulin-binding proteins within the slow phase of axonal transport in the rabbit vagus nerve.

Calmodulin-binding proteins (CBPs) in the rabbit vagus nerve were studied by means of calmodulin-Sepharose affinity chromatography and polyacrylamide gel electrophoresis. The soluble fraction (10(5) g supernatant) of a nerve homogenate contained four CBPs with molecular weights of 44, 55, 91, and 93 kD, respectively. Slowly transported proteins were recovered in the vagus 3 days after injection of [35S]methionine into the nodose ganglion. Four labelled CBPs with molecular weights of 44, 55, 69, and 83 kD, respectively were found. The nodose ganglion contained two labelled CBPs, 44 and 55 kD. The 55-kD CBP was identified as tubulin after immunoblotting. In separate experiments it was also shown that bovine brain tubulin bound to the calmodulin column.

Animals↗
PubMed2000

Analysis of mutants of tetanus toxin Hc fragment: ganglioside binding, cell binding and retrograde axonal transport properties.

Tetanus toxin binds neuronal tissue prior to internalization and trafficking to the central nervous system. Binding of the carboxy-terminal 50 kDa HC fragment of tetanus toxin to polysialogangliosides is important for this initial cell binding step. Using the three-dimensional structure of HC, mutants were designed to investigate the role of individual residues in ganglioside binding. Mutant proteins were tested for binding to GT1b gangliosides, to primary motoneurons and for their ability to undergo retrograde transport in mice. Two classes of mutant were obtained: (i) those containing deletions in loop regions within the C-terminal beta-trefoil domain which showed greatly reduced ganglioside and cell binding and did not undergo retrograde transport and (ii) those that showed reduced ganglioside binding, but retained primary neuronal cell binding and retrograde transport. The second class included point mutants of Histidine-1293, previously implicated in GT1b binding. Our deletion analysis is entirely consistent with recent structural studies which have identified sugar-binding sites in the immediate vicinity of the residues identified by mutagenesis. These results demonstrate that ganglioside binding can be severely impaired without abolishing cell binding and intracellular trafficking of tetanus toxin.

Animals↗
PubMed1999

Axonal transport of N-terminal huntingtin suggests early pathology of corticostriatal projections in Huntington disease.

Aggregation of N-terminal mutant huntingtin within nuclear inclusions and dystrophic neurites occurs in the cortex and striatum of Huntington disease (HD) patients and may be involved in neurodegeneration. We examined the prevalence of inclusions and dystrophic neurites in the cortex and striatum of 15 adult onset HD patients who had mild to severe striatal cell loss (grades 1, 2 or 3) using an antibody that detects the N-terminal region of huntingtin. Nuclear inclusions were more frequent in the cortex than the striatum and were sparse or absent in the striatum of patients with low-grade striatal pathology. Dystrophic neurites occurred in both regions. Patients with low-grade striatal pathology had numerous fibers with immunoreactive puncta and large swellings within the striatal neuropil, the subcortical white matter, and the internal and external capsules. In the globus pallidus of 3 grade 1 cases, N-terminal huntingtin markedly accumulated in the perinuclear cytoplasm and in some axons but not in the nucleus. Findings suggest that in the earlier stages of HD, accumulation of N-terminal mutant huntingtin occurs in the cytoplasm and is associated with degeneration of the corticostriatal pathway.

Adult↗
PubMed1980

Localization of parasympathetic preganglionic cell bodies innervating the pancreas within the vagal nucleus and nucleus ambiguus of the rat brain stem: evidence of dual innervation based on the retrograde axonal transport of horseradish peroxidase.

Traditionally studies have shown cell bodies of origin for parasympathetic preganglionic fibers of the subdiaphragmatic segment of the vagus nerve to be located in the dorsal motor nucleus of X. The nucleus ambiguus have been regarded as the motor nucleus for fibers projecting exclusively to larynx, pharynx and cervical esophagus. The results of the present study indicate that the pancreas has a dual source of innervation derived from both of these medullary nuclei, and that this in fact may be applicable for all or most of the subdiaphragmatic organs similarly innervated by the vagus.

Animals↗
PubMed1996

Glutamatergic connections of the auditory midbrain: selective uptake and axonal transport of D-[3H]aspartate.

D-[3H]aspartate was used to identify potential glutamatergic connections of the chinchilla inferior colliculus (IC). High-affinity uptake of D-[3H]aspartate is considered a selective marker for glutamatergic synapses, and neurons retrogradely labeled from such injections are believed to use glutamate, or a closely related compound, as a transmitter. Injections of D-[3H]aspartate suggest that glutamatergic endings in the IC arise primarily from intrinsic connections, the opposite IC, layer 5 of temporal cortex, nucleus sagulum, and lateral lemniscal nuclei. Neurons giving rise to the principal sensory (lemniscal) projections to the IC, i.e., those from the cochlear nuclei, superior olive, and the majority of projections from the lateral lemniscal nuclei, did not label in these experiments, indicating that their synapses do not recognize D-[3H]aspartate as a suitable substrate and may use inhibitory or other excitatory transmitters. After IC injections, fiber and diffuse labeling was found ipsilaterally in the medial geniculate body, superior colliculus, and dorsolateral pontine nuclei, contralaterally in the IC, and bilaterally in the superior olive and cochlear nuclei. Such labeling was attributed to anterograde transport of D-[3H]aspartate within the efferent collaterals of labeled IC neurons, suggesting that many of the IC's efferent projections may also be glutamatergic. This interpretation was confirmed in separate experiments in which D-[3H]aspartate, injected in the medial geniculate body, retrogradely labeled neurons in the IC as well as in layer 6 of temporal cortex. Finally, the mesencephalic trigeminal nucleus and tract labeled in some cases and may have local glutamatergic connections.

Animals↗
PubMed2000

The limbic zone of the rabbit and rat claustrum: a study of the claustrocingulate connections based on the retrograde axonal transport of fluorescent tracers.

The claustrum is a subcortical structure lying under the insular and piriform cortices, whose function is still not clear. Although data exist on connections of the claustrum and the limbic cortex, the topography of the limbic zone in the rabbit and rat claustrum has not been studied extensively. The study was performed on 17 adult Wistar rats and 12 New Zealand rabbits. Two percent water solutions of fluorescent retrograde tracers fast blue and nuclear yellow were injected into the various regions of the limbic cortex. The limbic zone is localized throughout the whole rostrocaudal extent of the claustrum, mainly in its ventromedial portion lying close to the external capsule. Although this zone of the claustrum is localized similarly in both rat and rabbit, some differences between these two species exist. In the rat, neurons projecting to all limbic areas are localized mainly in the anterior and central parts of the claustrum, whereas in the rabbit, the majority of the neurons projecting to the cingulate cortex are present in the anterior and central parts of this structure, while neurons sending axons to the retrosplenial cortex are localized in the central and posterior parts. In both species, double-labeling study showed that neurons projecting to various limbic regions are intermingled and that neurons sending axons into two different limbic regions are seen only occasionally. Our findings give support to the role of the claustrum in integrating information between different areas of the cerebral cortex and the limbic system.

Animals↗
PubMed1980

Projections from the visual cortex to the contralateral claustrum of the cat revealed by an anterograde axonal transport method.

Contralateral projections from visual areas 17, 18, 19 and the Clare-Bishop area of the cerebral cortex to the claustrum have been investigated in the cat using intracortical injections of [3H]proline. Radioactive material was found in a dorsocaudal region of the contralateral claustrum. This region was homotopic with respect to that found for the ipsilateral projection from visual cortex. The contralateral connection is assumed to be a monosynaptic pathway. The pattern by which the corticofugal fibres terminate in the claustrum is quite similar to the one described for the opposite hemisphere [6].

Animals↗
PubMed1983

Morphological alterations of retinal cells accompanying inhibition of axonal transport by kainic acid.

The effect of kainic acid (KA) on rat retinal morphology was investigated with emphasis on the ganglion cell layer. Two hours after intraocular injection of KA, 70% of the neurones in the ganglion cell layer display morphological alterations. These alterations at the ultrastructural level include the absence of identifiable Golgi complexes and the presence of abundant round empty vacuoles among other nuclear and cytoplasmic modifications. These alterations of the Golgi complex are discussed in relation to axoplasmic transport inhibition by kainic acid. Seven days after KA treatment the retina showed a decreased number of cells in the inner nuclear layer and ganglion cell layer and the inner plexiform layer had virtually disappeared. Despite the disappearance of cells from the ganglion cell layer a week after KA injection, abundant ganglion cells could be retrogradely labelled with horseradish peroxidase. Thus although retinal ganglion cells seem to be sensitive to KA most of them survive KA treatment.

Animals↗
PubMed1994

Processing, axonal transport and cardioregulatory functions of peptides derived from two related prohormones generated by alternative splicing of a single gene in identified neurons VD1 and RPD2 of Lymnaea.

The VD1/RPD2 mRNA precursor in identified neurons VD1 and RPD2 of the freshwater snail Lymnaea stagnalis is alternatively spliced to yield two related variants encoding two distinct yet related preprohormones, named the VD1/RPD2-A and -B preprohormones. Here, we report the isolation and structural characterization of alpha 1, alpha 2 and beta peptides from dissected neurons VD1 and RPD2. The alpha 1 and alpha 2 peptides are derived from VD1/RPD2-A and B prohormones, respectively, whereas beta peptide is identical for both prohormones. In addition, we report the isolation and structural characterization of the alpha 2 peptide from the heart, demonstrating that the mature peptides are transported and released in the heart. The pharmacological actions of synthetic alpha 1 and alpha 2 peptides on isolated auricle preparations of the Lymnaea heart were examined. The two alpha peptides have similar excitatory effects on beat rate and beat amplitude, while their potencies differed considerably, indicating that alternative splicing results in structurally and functionally overlapping, through non-identical, sets of peptides.

Amino Acid Sequence↗
PubMed1987

Use of muscle contraction formalism for kinesin in fast axonal transport.

The general procedure is discussed for calculating the velocity of a vesicle along a microtubule. The formalism used previously for isotonic contraction in muscle (with multiple actin sites for a given cross-bridge) can be employed. However, some modifications must be made: (i) the kinetic diagram must include a state in which kinesin is absent from a vesicle binding site, (ii) an average must be taken over the locations of the vesicle binding sites relative to microtubule sites, and (iii) a self-consistency condition must be imposed that equates the mean force exerted by kinesin molecules on the vesicle with the frictional resisting force of the medium.

Animals↗