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Biomedical subjects

R E Snyder

Publications and source records attributed to R E Snyder.

At least 37 records · Page 2Linked to original sources
PubMed1991

Reversal of rapid axonal transport at a lesion: leupeptin inhibits reversed protein transport, but does not inhibit reversed organelle transport.

The hypothesis that the reversal of rapid axonal transport requires a proteolytic conversion of anterograde transport vesicles was examined using sciatic nerve preparations from Xenopus laevis and leupeptin as an inhibitor of proteolysis. The transport of newly synthesized 35S-labeled proteins was studied with a position-sensitive detector of radiation. Organelle transport in isolated myelinated axons was studied by video microscopy. Leupeptin (0.1-0.4 mM) reduced the anterograde-to-retrograde reversal of protein transport adjacent to an axonal lesion. In experiments in which organelle transport was observed close to lesions in axons maintained in a medium compatible with intracellular function, 1.0 mM leupeptin inhibited neither the anterograde-to-retrograde nor the retrograde-to-anterograde reversal of organelle transport. In addition, in experiments in which conditions approximated those used to study protein transport, organelle transport away from the lesion was not inhibited by 1.0 mM leupeptin. A comparison of the morphology of rapidly transported organelles that underwent anterograde transport to the morphology of those that returned from a lesion (with or without the presence of leupeptin) provided no evidence that a morphological conversion was a necessary step in transport reversal.

Animals↗
PubMed1989

Junction between parent and daughter axons in regenerating myelinated nerve: properties of structure and rapid axonal transport.

The primary aim of this work was to investigate the properties of rapid axonal transport in regenerating myelinated axons in the sciatic nerve of Xenopus laevis, with particular attention to events at the junction between the proximal, intact axon (the "parent") and the distal, newly formed axon (the "daughter"). Morphological studies indicated that all myelinated axons initiated regeneration and that at least 80% of these axons regenerated at a rate of 1 mm/day or greater (20 degrees C). The ultrastructure of the junctional region was examined at regeneration times between 3 days and 20 weeks. The main qualitative change in the junctional axoplasm over this period was in its content of particulate organelles. At times up to 2 weeks regeneration, the junction contained abnormal numbers of 50 nm diameter vesicles and 10 nm granules. Between 2 and 5 weeks the junction showed in addition a peripheral rim of large membrane-bounded organelles around a central core of microtubules and neurofilaments. At longer times the numbers of large membrane-bounded organelles diminished and all junctions contained prominent accumulations of 10 nm granules. The rate of rapid axonal transport of protein was similar in parent and daughter axons. Compared to the parent axons, a 2-5 times greater amount of protein was deposited to a stationary phase in daughter axons. Specimens of nerve that were subjected to mechanical stress during the removal of the perineurium showed a large accumulation of rapidly transported protein in the region of the crush at regeneration times up to 40 days; some of the accumulated protein was subsequently transported retrogradely. Video microscopy of isolated axons supplied evidence that the transport deficit in mechanically stressed nerve was a partial block of anterograde vesicle transport, plus a reversal of anterograde transport, at the junction of parent with daughter axons. No structural changes were detected in mechanically stressed nerve. The results show that the junction between parent and daughter myelinated axons is a region with distinct morphology at which the dynamics of anterograde axonal transport may change dramatically.

Animals↗
PubMed1989

Loss of material from the retrograde axonal transport system in frog sciatic nerve.

Rapid axonal transport was studied in sciatic nerve preparations of the amphibian Xenopus laevis maintained in vitro at 23.0 +/- 0.2 degrees C. A pulse of [35S]methionine-labeled material was allowed to move in the anterograde direction until encountering a lesion, at which a portion of the pulse reversed directions and moved in the retrograde direction. By constricting the nerve during the course of the experiment, it was possible to prevent continuous return of label from the lesion, thus creating a retrogradely moving pulse that contained a defined quantity of radiolabel. Movement of both the anterograde and the retrograde pulse were monitored continuously for up to 24 h using a position-sensitive detector of ionizing radiation. The front and the back edge of the anterograde pulse were found to move at the rates of (mm/day) 179.9 +/- 3.9 (+/- SEM) and 149.9 +/- 5.9, respectively, and the front and the back edge of the retrograde pulse moved at the rates of 155.8 +/- 11.3 and 84.6 +/- 2.9, respectively. By comparison of the quantity of label lost to the stationary phase to the quantity of label calculated to have been present in the anterograde pulse, it was determined that 0.068 +/- 0.009 of the anterograde pulse is lost to each 3.18-mm region of nerve. Comparison of the quantity of label calculated to have been present in the retrograde pulse to that in the anterograde pulse revealed that 0.057 +/- 0.014 of the retrograde pulse is lost to each 3.18-mm region of nerve. It is concluded that protein originating in the cell body and which reverses its direction of transport at a lesion can be lost from the retrograde axonal transport system.

Animals↗
PubMed1988

The release of axonally transported material from an in vitro amphibian sciatic nerve preparation.

The rapid axonal transport of a pulse of [35S]methionine-labelled material was used to study the release of transported material from amphibian nerve maintained in vitro. Following creation of a moving pulse of activity in a dorsal root ganglion-sciatic nerve preparation, the ganglion was removed and the nerve placed in a three-compartment tray, the section of nerve in the middle compartment containing no truncated branches (unbranched section). All three compartments were filled with a saline solution that in some studies contained nonradioactive methionine (1.0 mmol/L). Analysis of studies in which nonradioactive methionine was absent revealed that labelled material appeared in the bathing solution of the end compartments that contained truncated branches, but not in the solution of the middle (unbranched) compartment. The quantity of label released in the branched compartments was approximately 6% of that remaining in the corresponding section of nerve following an 18-20 h incubation period. However, when nonradioactive methionine was present, all compartments showed an additional activity in the bathing solution of approximately 10% of that remaining in the nerve. In another study in which a position-sensitive detector of ionizing radiation was used to monitor progress of the pulse, it was found that activity did not enter the bathing solution of a compartment prior to the pulse of activity. It is concluded that in the absence of methionine from the bathing solution, axonally transported material is released only from regions of nerve that contain severed axons; however, the presence of methionine allows transported material to be released from nerve containing intact axons. Ultrafiltration studies and thin-layer chromatography revealed the majority of material released to be of low-molecular weight (less than 30,000 daltons) and not free [35S]methionine.

Animals↗
PubMed1986

The kinematics of turnaround and retrograde axonal transport.

Rapid axonal transport of a pulse of 35S-methionine-labelled material was studied in vitro in the sensory neurons of amphibian sciatic nerve using a position-sensitive detector. For 10 nerves studied at 23.0 +/- 0.2 degrees C it was found that a pulse moved in the anterograde direction characterized by front edge, peak, and trailing edge transport rates of (mm/d) 180.8 +/- 2.2 (+/- SEM), 176.6 +/- 2.3, and 153.7 +/- 3.0, respectively. Following its arrival at a distal ligature, a smaller pulse was observed to move in the retrograde direction characterized by front edge and peak transport rates of 158.0 +/- 7.3 and 110.3 +/- 3.5, respectively, indicating that retrograde transport proceeds at a rate of 0.88 +/- 0.04 that of anterograde. The retrograde pulse was observed to disperse at a rate greater than the anterograde. Reversal of radiolabel at the distal ligature began 1.49 +/- 0.15 h following arrival of the first radiolabel. Considerable variation was seen between preparations in the way radiolabel accumulated in the end (ligature) regions of the nerve. Although a retrograde pulse was seen in all preparations, in 7 of 10 preparations there was no evidence of this pulse accumulating within less than 2-3 mm of a proximal ligature; however, accumulation was observed within less than 5 mm in all preparations.

Animals↗
PubMed1986

A radiolabelled pulse for the simultaneous study of anterograde and retrograde axonal transport.

A technique is described for producing a pulse of [35S]methionine-labelled material which is axonally transported in amphibian sciatic nerve maintained in vitro. Using a position-sensitive detector of ionizing radiation, it is possible to continuously observe the movement of the pulse in the anterograde direction and, following turnaround at a ligature, the movement of a fraction of the pulse in the retrograde direction. Two sources of contaminant activity, which would otherwise interfere with observation of the pulse, are discussed and shown to be avoidable.

Animals↗
PubMed1985

Rapid axonal transport in Xenopus nerve in divalent cation free media.

An investigation was made of the effects of bathing media low in divalent cations on rapid axonal transport in the sciatic nerve of the amphibian Xenopus laevis. The anterograde transport of a pulse of [35S]methionine proteins was observed using a multiple proportional counter as the detector. Organelles undergoing anterograde and retrograde transport were detected by light microscopy. The structure of nerve fibers was examined by light and electron microscopy. There was no significant difference in the anterograde transport of proteins in nerves bathed in normal medium (NM) containing millimolar Ca2+ and Mg2+ and in those bathed in calcium-free medium (CaFM) containing Mg2+. The anterograde transport of labelled proteins continued at a normal velocity in nerves bathed in divalent cation free medium (DCFM) for at least 14 h. DCFM did cause some alterations in protein transport: the ratio of the plateau (following pulse passage) to the peak radioactivity was increased, the pulse amplitude decreased more rapidly, and the label continued to arrive at the distal end of the nerve for greater than 16 h. Anterograde and retrograde organelle transport continued normally for periods of greater than or equal to 4 h in fibres bathed in DCFM. All myelinated fibres became distorted within 4 h in DCFM. Similar distortion was rare in fibres bathed in CaFM. The results indicate that axonal transport in Xenopus is largely independent of lowered concentrations of divalent cations in the bathing medium. Those alterations in axonal transport that were produced by DCFM may have been secondary to morphological changes in the nerve fibres.

Animals↗
PubMed1984

A temporal variation in nonneuronal protein synthesis in dorsal root ganglia and nerve and its significance to studies of axonal transport.

Protein synthesis and fast axonal transport were studied in vitro using dorsal root ganglia (DRG)-sciatic nerve preparations from the amphibian Xenopus laevis. It was observed that the rate of incorporation of [3H]leucine into protein in DRG and isolated segments of nerve began to increase 9 to 11 h after killing the animal, attaining at 13 to 17 h a maximum of 5- to 10-times preincrease (less than 9 h) values. At the same time as an increase in the rate of incorporation began, synthesis commenced in DRG and nerve exposed to cycloheximide (125 micrograms/ml). Whereas cycloheximide reduced fast axonal transport to 1 to 3% of control values in preparations maintained 20 to 24 h in vitro, cycloheximide reduced incorporation in DRG to only 80% of control values. N-terminal labeling studies showed that both the increased incorporation and cycloheximide-insensitive incorporation resulted from protein synthesis. Autoradiographic and incorporation studies indicated that nonneuronal cells situated in the ganglion capsule and perineural sheath of the nerve were responsible for both the increased incorporation and cycloheximide-insensitive synthesis. The findings have implications for the study of axonal transport.

Animals↗
PubMed1984

Physical methods for the study of the dynamics of axonal transport.

Currently used techniques for the study of the dynamics of axonal transport are reviewed. Emphasis is placed on those nonstructural and nonbiochemical methods which are most useful in the investigation of the dynamics of the mechanisms that underline the transport process. The methods by which the transport of specified materials may be assessed are divided into two broad groups: those which involve the collection, or accumulation, of transported materials at a region where transport is slowed or arrested, and those methods in which the distribution of a marker of the transported material is assayed in a series of nerve segments. Potential problems and sources of error associated with these techniques are considered as is their applicability to particular biological preparations. Recently introduced methods for concentrating and labeling transported material are described. New methods for conducting segment analysis of transported material by the use of position-sensitive detectors of radiation are considered. Optical methods of detecting axonal transport are reviewed with emphasis being placed on recently introduced methods and on the means by which the motion of optically detected organelles may be analyzed. In addition, biological preparations which are particularly suited to the optical approach are described.

Animals↗
PubMed1983

Calcium efflux from amphibian sciatic nerve.

Eight Xenopus laevis were injected intraperitoneally with 45CaCl2 and 16-18 h later an unbranched section from each sciatic nerve was removed. Efflux measurements of nerve from which the perineurial sheath had been removed could be described by three compartments of approximately equal size with half-lifes of 2.37 +/- 0.76 (SD), 30.3 +/- 17.3 and 196 +/- 61 min, the shortest lived compartment representing diffusion from the extracellular space with a coefficient of diffusion of 2.1 +/- 0.7 X 10(-6) cm2/s. Efflux from nerve in which the perineurium remained intact was characterized by a half-life of 862 +/- 399 min resulting from the sheath acting as a diffusion barrier of permeability 3.4 +/- 1.6 X 10(-7) cm/s. The perineurium was found to bind or sequester a quantity of calcium 1-2 times that contained in an equal volume of plasma.

Animals↗
PubMed1982

Position-sensitive detector studies of the axonal transport of a pulse of radioisotope.

Axonal transport of a pulse of 35S-methionine-labelled material was studied in vitro in amphibian sciatic nerve using position sensitive detectors. Following formation of a pulse of activity using the cold block technique, the nerve was ligated proximal and distal to the pulse and its movement monitored at room temperature (22.5-23.5 degrees C) for up to 16 h. Material transported in the anterograde direction did so with an average maximum velocity of 147 mm/d. The pulse was found to disperse at an average rate of 0.23 mm/mm travel; however, dispersion was found to vary from preparation to preparation more than would be predicted from experimental error alone. Label was observed to reverse direction at the distal ligature in only 2 of 13 preparations. Reversal of label began within approximately 0.4 h of first arrival, and the most rapidly retrogradely transported material moved at a velocity of 80% that of the most rapidly anterogradely transported material.

Animals↗
PubMed1982

The in situ measurement of strontium content in bone using x-ray fluorescence analysis.

A technique is described which uses x-ray fluorescence to determine in situ the strontium content of bone. Measurements were made of the skulls of seven rabbits both before and after a single bolus injection of strontium was administered intravenously. The retention of strontium was followed for up to eight months and could be represented either by a three-exponential model or, for data obtained between 12 and 252 d following injection, the power function At-b, where t is the time in days following injection. The three-exponential terms had linear coefficients of similar magnitude and half lives of 3-4 h, 10-40 d and 200-300 d. The exponent of the power function was 0.25-0.57. Measurements were also made of the skull and medial surface of the tibial crest of an adult dog for four months following subcutaneous administration of strontium. The results obtained using x-ray fluorescence are found to be similar to those reported by others for radiostrontium studies. The precision and errors associated with the fluorescence technique are discussed. Estimates of radiation dose are given. It is concluded that the technique may serve to determine in situ the strontium content of bone near the surface of the body.

Animals↗
PubMed1982

The action of puromycin and cycloheximide on the initiation of rapid axonal transport in amphibian dorsal root neurones.

1. Amphibian dorsal root ganglia-sciatic nerve preparations were incubated in vitro and the rapid axonal transport of radioactive labels was studied with a position-sensitive detector and by conventional liquid scintillation analysis. Protein was labelled by exposure of the ganglia to [(35)S]methionine or [(3)H]leucine and lipid was labelled using [(32)P]orthophosphoric acid.2. Protein synthesis was interrupted by exposure of the ganglia to either cycloheximide or puromycin. When ganglia were exposed to either inhibitor prior to or simultaneously with a label, the somal export of both protein and lipid to the axon was reduced by two to three orders of magnitude.3. Using the position-sensitive detector, [(35)S]methionine was observed to be exported from the ninth dorsal root ganglia of Rana catesbiana 3.49+/-1.56 h (+/- S.D.) after exposure, and [(32)P]phosphate 4.46+/-1.85 h after exposure.4. Export of [(35)S]methionine or [(32)P]phosphate was disrupted 3.32+/-1.21 h (+/- S.D.) or 1.93+/-1.04 h respectively after exposure of the ganglia to cycloheximide or puromycin.5. For a given preparation the time required for [(35)S]methionine to be exported was statistically equal to the time required for cycloheximide or puromycin to disrupt export. No such correlation was found to exist for the export of [(32)P]phosphate.6. Analysis revealed that materials labelled with either [(35)S]methionine or [(32)P]phosphate continue to be exported from the ganglia for several hours after the initial disruption in outflow caused by the inhibitors.7. The results do not provide support for the hypothesis of Ambron, Goldman & Schwartz (1975) that a ;key' newly synthesized, and non-storable, polypeptide is added to an already assembled structure to allow rapid axonal transport to be initiated.

Animals↗
PubMed1980

Epidemiology of breast carcinoma II: factors related to the predominance of left-sided disease.

A study of the laterality of 980 patients with unilateral breast carcinoma revealed a left/right ratio of 1.26. Detailed analysis disclosed a significant association between left predominance and the following clinicopathologic features: menarch after age 13, age at diagnosis, parity especially among those between ages 40 and 54, and all histologic types except medullary tumors. When asynchronous bilateral carcinoma was documented, the disease first occurred more often in the left breast. Patients with simultaneous bilateral disease usually had a larger tumor on the left. Because differences in breast size were considered a contributing factor, relative breast volumes were computed from the mammograms of 174 healthy women; 55% were found to have a larger left breast. These data and other observations cited in the literature suggest that the asymmetry of breast carcinoma reflects differences in the sensitivity of the mammary glands to hormonal stimulation, resulting in unequal volumes of tissue at risk to develop carcinoma.

Adolescent↗
PubMed1980

Specimen radiography and preoperative localization of nonpalpable breast cancer.

Specimen radiography with or without needle localization should become standard procedure when breast biopsy is performed solely on the basis of mammographic findings. Without these techniques, surgery may prove useless. Over a five-year period from 1974 through 1978, a series of 423 specimen radiographs were done at Memorial Sloan-Kettering Cancer Center for questionable mammographic calcifications or, in a few instances, nonpalpable masses. One hundred and twenty-six proved to be carcinoma, but they represent only 4% of the total number of cancers treated during this period of time. Most breast cancers are still being detected by palpation or palpation combined with mammography. The cancers needing specimen radiography or needle localization are the truly minimal lesions with the best prognosis. With present day techniques, radiation dosage from mammography is extremely low. Its continued use should be encouraged if mammography is to have any impact on breast cancer survival rates. High quality mammograms, interpretation by experienced radiologists, and close coordination of the efforts of the radiologists, surgeon, and pathologist can make possible the detection of increasing numbers of these minimal cancers.

Biopsy↗
PubMed1980

A clinicopathologic study of atypical lesions of the breast further follow up.

On the assumption that both the lobular and intraductal atypical lesions of the breast are precancerous in nature, a group of 572 such patients have been closely followed during the past 12 years. Thirty-four patients eventually developed cancer during this period. The cumulative incidence of breast cancer was 7.74% at 10 years in the group of patients with atypical lesions only and 16.15% at 10 years in the patients with atypical lesions in one breast and cancer in the contralateral breast. The transformation of this probable precancerous state into cancer apparently takes a long interval of time.

Adolescent↗
PubMed1980

Rapid orthograde transport of 32P-labelled material in amphibian sensory axons: a multiwire proportional chamber study.

A multiwire proportional chamber was used to follow the axonal transport of material labelled with [32P]orthophosphate in dorsal root ganglion (DRG)--sciatic nerve preparations of Xenopus laevis and Rana catesbiana. The DRG were exposed to label for a period of 4 h following which there was a period of continued delivery of labelled material to the nerve for up to 18 h. The front of the labelled material in the nerve moved at a velocity of 160--170 mm/24 h at room temperature (22.5--23.5 degrees C). Sectioning the nerve at a proximal position showed that labelled material behind the front moved at a similar rapid velocity. Experiments in which the nerve was sectioned showed that some of the rapidly transported label appeared to be deposited into a relatively stationary phase. Extrapolation of the results indicated that the delay between the presentation of the label to the DRG and the onset of the transport of labelled material in the nerve was 4--6 h. The rapid transport of the label was inhibited by vinblastine sulphate at concentrations of 130--950 microM. Most of the rapidly transported material was found to be in a chloroform-methanol extractable form. In conclusion, 32P labels materials whose transport dynamics are very similar to those observed when [35S]methionine is used as the precursor.

Animals↗