Tannic acid-stained microtubules with 12, 13, and 15 protofilaments

Burton, P R; Hinkley, RE; Pierson, G B

doi:10.1083/jcb.65.1.227

Cited by 111 publications

(54 citation statements)

References 21 publications

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“…Third, axonal MTs have a uniform polarity orientation, with their plus ends pointing away from the cell body toward the axon tip (Heidemann et al, 1981;Burton and Paige, 1981). Finally, axonal MTs have a 13 protofilament substructure (Tilney et al, 1973;Burton et al, 1975).…”

Section: Abstract: Axon Outgrowth; Cultured Sympathetic Neurons; Micmentioning

confidence: 99%

Microtubule Transport from the Cell Body into the Axons of Growing Neurons

1997

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The present studies test the hypothesis that microtubules (MTs) are transported from the cell body into the axons of growing neurons. Dissociated sympathetic neurons were cultured using conditions that allow us to control the initiation of axon outgrowth. Neurons were injected with biotin-labeled tubulin (Bttub) and then stimulated to extend axons. The newly formed axons were then examined using immunofluorescence procedures for MTs with or without Bt-tub. Because the Bt-tub is fully assembly competent, all MTs that assemble after injection will contain Bt-tub. However, MTs that exist in the neuron at the time of injection and persist during the subsequent incubation will not contain Bt-tub. Because the neurons were injected before extending axons, MTs without Bt-tub are initially localized to the cell body. We specifically determined whether these MTs appeared in the newly formed axon. Such a result can only be explained by the transport of these MTs from their initial location in the cell body into the axon. The newly formed axons of many neurons contained MTs both with and without Bt-tub. MTs without Bt-tub were detected all along the axon and in some neurons represented a substantial portion of the total polymer in the proximal and middle regions of the axon. These results show that MTs are transported from the cell body into growing axons and that this transport is robust, delivering MTs to all regions of the newly formed axon.

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Section: Abstract: Axon Outgrowth; Cultured Sympathetic Neurons; Micmentioning

confidence: 99%

Microtubule Transport from the Cell Body into the Axons of Growing Neurons

1997

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“…Within the axon, the MT array is continuous from the cell body into the terminal growth cone, but individual MTs vary in length, stopping and starting at various points within the array (Bray and Bunge, 1981;Yu and Baas, 1994). All of the MTs have a consistent 13-protofilament lattice (Tilney, 1973;Burton et al, 1975), and are uniformly oriented with regard to their intrinsic polarity, with plus ends directed away from the cell body (Heidemann et al, 198 1;Baas et al, 1988). Although it has been long recognized that axonal MTs are not attached to any discernable nucleating structure such as the centrosome (Lyser, 1968), contemporary studies suggest that axonal MTs are in fact nucleated at the centrosome, after which they are released for transport into the axon (Baas and Joshi, 1992;Baas and Ahmad, 1993;Yu et al, 1993;Ahmad et al, 1994).…”

Section: Serial Reconstruction Analyses Indicate That Mt Number Incrementioning

confidence: 99%

Microtubule fragmentation and partitioning in the axon during collateral branch formation

1994

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Axons within the brain branch principally by the formation of collaterals rather than by bifurcation of the terminal growth cone (O'Leary and Terashima, 1988). This same behavior is recapitulated in cultures of embryonic hippocampal neurons (Dotti et al., 1988), rendering them ideal for studies on the cell biological mechanisms underlying collateral branch formation. In the present study, we focused on changes in the microtubule (MT) array that occur as these axons branch. In particular, we explored the mechanism by which MT number is locally increased to accommodate the need for more MTs during collateral branch formation. Serial reconstruction analyses indicate that MT number increases by severalfold and that MT length decreases correspondingly within the parent axon in the discrete region giving rise to the branch. These observations strongly suggest that MTs within the parent axon undergo a local fragmentation in this region, and hence raise the possibility that a portion of these new MTs might be destined for transport into the branch. To address this latter issue, we used quantitative immunofluorescence to compare the proportion of newly assembled to total MT polymer in different regions of the axon. As previously reported (Brown et al., 1992), the region of the axon contiguous with the terminal growth cone is particularly rich in newly assembled polymer. In contrast, there was no distinguishable difference in the proportion of newly assembled polymer in the newly formed collateral branches compared to the shaft region of the parent axon. These results indicate that the MTs within the newly formed collateral branches are on average assembled at the same time as those within the parent axon, and thus strongly suggest that the MTs in the collateral branch were assembled in the parent axon and then translocated into the branch. We conclude on the basis of these observations that collateral branch formation requires a local fragmentation of MTs within the parent axon, followed by the partitioning of a portion of the MT fragments into the branch. These short MTs presumably then resume their movement and elongation down the collateral branch as well as down the parent axon for the steady and orderly increase of both MT arrays.

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“…The fixatives containing tannic acid have been used to demonstrate the structures of microtubules [Fuiaesakn el a'., 1972;Mizuhira and Futaesaku, 1972;Tilney el a/., 1973;Burton et a!., 1975], intercellular junctions [ Van Dears, 1975], capillary endothelial cell membranes [IVagner, 1976] and T-tubules of skeletal muscle [Bonilla, 1977], In respect to the structural relationship of muscle cell membranes, collagen libers and microfibrils, ihe authors examined electron microscopically the myotendon junction of bull frog muscles using tannic acid as the fixative.…”

Section: Introductionmentioning

confidence: 99%

Microfibrils in the myotendon junctions

Ajiri¹,

Kimura²,

Ito³

et al. 1978

Cells Tissues Organs

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Myotendon junctions in the rectus abdominis muscles of bull frogs were examined by the fixation combination of tannic acid and glutaraldehyde using electron microscopy. The features observed on myotendon junctions were the following: (1) There were many deep invaginations of muscle cell membrane at the end of the muscle fibers. Terminal thin filaments of myofibrils were attached to the electron-dense layer lining under the muscle cell membrane on the lateral walls of invaginations. (2) The basement membrane covering the muscle cell membrane was thicker in the invaginations than on the other sites of muscle fibers. (3) Collagen fibers in the invaginations gradually tapered off toward the bottom of the invaginations. But it was not seen that the collagen fibers were attached to both the basement membrane and cell membrane of muscle cells. (4) On the observations using the tannic acid-glutaraldehyde fixation, it was clearly seen that the microfibrils extend from the outer leaflets of the cell membrane to the collagen fibers in invaginations via the basement membrane. It was concluded that the myofibrils might be fastened to the collagen fibers of the tendon by the intermediates of the microfibrils.

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Tannic acid-stained microtubules with 12, 13, and 15 protofilaments

Cited by 111 publications

References 21 publications

Microtubule Transport from the Cell Body into the Axons of Growing Neurons

Microtubule Transport from the Cell Body into the Axons of Growing Neurons

Microtubule fragmentation and partitioning in the axon during collateral branch formation

Microfibrils in the myotendon junctions

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