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Biomedical subjects

R D Fields

Publications and source records attributed to R D Fields.

51 records · Page 3Linked to original sources
PubMed1988

Regional membrane heterogeneity in premyelinated CNS axons: factors influencing the binding of sterol-specific probes.

Binding of the sterol-specific probe filipin to developing optic nerve axonal membrane is spatially heterogeneous prior to association of glial cells with the axons. Experiments were performed using different sterol binding probes (filipin, tomatin, and saponin), at different temperatures (4 degrees C, 23 degrees C, and 37 degrees C), after incubation in different ionic conditions (10 mM Ca2+, 10 mM EGTA, and 20 mM Mg2+), to examine factors that may be responsible for this membrane heterogeneity in rat optic nerve. The patchy pattern of filipin binding is apparent with each sterol-specific probe, even prior to glial ensheathment, and is retained when membrane fluidity is increased at higher temperatures. Increased Ca2+ concentration increased membrane stability, and increased Mg2+ reduced the patchiness of filipin binding. After tannic acid staining, regions of the cytoskeleton are seen associated with the membrane via filaments extending from microtubules to the membrane, preferentially in regions where filipin interaction with the membrane is inhibited. The non-uniform interaction of filipin with the axolemma suggests an underlying heterogeneity in the sterol composition and stability of the membrane. Heterogeneity of premyelinated axonal membrane may provide an important formative influence in the differentiation of axons to their mature morphology and function.

Animals↗
PubMed1988

Diminished dorsal root GABA sensitivity following chronic peripheral nerve injury.

The depolarizing effect of gamma-aminobutyric acid (GABA) on rat lumbar dorsal roots was studied in a sucrose gap chamber following axotomy or crush injury of the sciatic nerve or dorsal root. The mean depolarization elicited by GABA on normal dorsal roots (3.96 +/- 0.71 mV, N = 14) was significantly reduced following chronic sciatic axotomy (2.02 +/- 0.99 mV, N = 15). Chronic sciatic crush injury had no significant effect on dorsal root GABA sensitivity. The amplitudes of the dorsal root compound action potentials were the same from rats with normal and injured sciatic nerves, indicating that axons proximal to the sciatic nerve lesion did not undergo appreciable degeneration. A marked loss of dorsal root GABA sensitivity was also seen following dorsal root axotomy or crush injury (1.02 +/- 0.98 mV (N = 10) and 0.69 +/- 0.70 mV (N = 9), respectively). These results indicate that GABA sensitivity of dorsal roots is attenuated following peripheral nerve lesions in which regeneration and functional reconnection with peripheral targets are prevented. Previous work indicates that the primary afferent depolarization is reduced under similar conditions. The reduction in GABA sensitivity of dorsal root fibers described here may have a contributory role in the reduced primary afferent depolarization that follows peripheral nerve transection, which has pathophysiologic implications in chronic pain syndromes.

Action Potentials↗
PubMed1988

Functionally significant plasticity of synaptic morphology: studies on the ribbon synapse of the ampullae of Lorenzini.

Changes in electrophysiological properties measured in vitro were correlated with ultrastructural differences at synapses between sense cells and the primary afferent neurons in electrosensory organs of the thornback ray (the ampullae of Lorenzini). Variation in synaptic structure was classified into four synaptic morphotypes, which appear to represent stages in a cyclic pattern of ultrastructural modification associated with changes in synaptic efficacy. Synapses with deeper postsynaptic troughs, and active zone regions located at the "narrow point" of the presynaptic evagination, and other morphological differences, were associated with greater sensitivity and spontaneous activity. Furthermore, the morphology of synapses was different in organs that had shown increasing, decreasing or stable trends in sensitivity prior to fixation, suggesting that changes in synaptic physiology and morphology are interrelated, and providing evidence for the sequence of ultrastructural modifications represented by the four synaptic morphotypes. These results support the conclusion that synaptic morphology is plastic and that this plasticity has functional significance in terms of the threshold sensitivity and spontaneous activity monitored from the afferent nerves. Plasticity of synaptic morphology which is associated with changes in the efficacy of transmitter release at chemically mediated synapses could be important in relatively long-term phenomena.

Animals↗
PubMed1988

Macromolecular structure of axonal membrane in the optic nerve of the jimpy mouse.

The macromolecular structure of the axon membrane in 26-28-day-old Jimpy mice and control optic nerve were examined with quantitative freeze-fracture electron microscopy. Premyelinated and myelinated axons were observed in control optic nerves, with axonal diameters of premyelinated axons being generally smaller than that of myelinated axons (approximately 0.2-0.4 micron vs approximately 0.5-1.5 micron, respectively). Axon membrane from control optic nerves exhibited an asymmetrical partitioning of intramembranous particles (IMP). P-faces of internodal membrane displayed nearly twice as many IMP as the premyelinated axolemma (1,731 vs 893 micron-2, respectively). E-faces of internodal and premyelinated axolemma exhibited IMP densities of 124 and 157 micron-2, respectively. Few myelinated axons were apparent in optic nerves from Jimpy mice. The amyelinated axons of Jimpy mice displayed a spectrum of axonal diameters, ranging from approximately 0.2 to 1.5 micron. P-face densities of amyelinated axons, considered as a group, exhibited a wide range (600-2,100 micron-2). However, large diameter (greater than or equal to 0.5 micron) axons exhibited a significantly greater P-face IMP density than that of small caliber (greater than 0.5 micron) axons (1,525 vs 1,032 micron-2, respectively). Aggregations of E-face IMP were not observed along amyelinated axons of Jimpy optic nerves. The results demonstrate that the changes in P-face IMP density that occur during development of normal myelinated axons also occur in developing axons of Jimpy optic nerve, irrespective of a lack of normal glial cell association, and provide further evidence that the primary defect of hypomyelination within Jimpy mice is not attributed to the neuron.

Animals↗
PubMed1987

Filipin-cholesterol binding in CNS axons prior to myelination: evidence for microheterogeneity in premyelinated axolemma.

The distribution of cholesterol in axonal membrane of developing rat optic nerves prior to myelination was studied by freeze-fracture cytochemistry. Binding of the cholesterol-specific probe, filipin, to the axolemma of premyelinated axons was heterogeneous; this suggests the presence of microdomains of axolemma with different membrane composition and/or cytoskeletal/extracellular matrix association. Although the reasons for this binding pattern have not yet been determined, heterogeneity occurs prior to association of glia with the axon, and may reflect regional differences in lipid/sterol composition of the axonal membrane bilayer, or distribution of membrane-associated cytoskeleton. The distribution of intramembranous particles was not obviously associated with the pattern of filipin binding in early developing axons, however, as might have been expected from the attending differences in fluidity of the membrane microdomains. Microheterogeneity in axonal membranes of developing axons could have an influence on several membrane properties, and may be associated with processes important for growth and differentiation of axons.

Aging↗
PubMed1987

Changes in synaptic morphology associated with presynaptic and postsynaptic activity: an in vitro study of the electrosensory organ of the thornback ray.

The influence of synaptic activity on synaptic structure was studied by selectively stimulating the presynaptic or postsynaptic membranes of ribbon synapses in an in vitro preparation, and examining the ultrastructure of synapses with conventional electron microscopic methods. Functionally significant changes in synaptic morphology were observed after direct depolarization of the presynaptic membrane or incubation with the neurotransmitter glutamate to depolarize the postsynaptic membrane. After depolarizing the presynaptic membrane for 30 seconds, the depth of the postsynaptic trough was reduced, and other morphological changes correlated with decreased sensitivity and spontaneous activity were evident. Depolarizing the postsynaptic membrane by incubating synapses with the neurotransmitter glutamate, produced opposite effects. These results suggest that synapses can undergo functionally significant morphological changes in response to certain patterns of activity. The mechanism for these changes might include synaptic vesicle recycling processes, changes in ion concentration, or cytoskeletal alterations in the presynaptic, postsynaptic, or support cells. These mechanisms could operate in association with long-term changes in synaptic efficacy or account for some physiological phenomena such as synaptic fatigue or accommodation.

Action Potentials↗
PubMed1986

Differences in intramembranous particle distribution in the paranodal axolemma are not associated with functional differences of dorsal and ventral roots.

Freeze-fracture methods were used to study intramembranous particles, which are believed to represent ectopic sodium channels in the paranodal axolemma, and their possible association with differences in action potential electrogenesis of maturing rat dorsal and ventral root fibers. Our results indicate that there is no significant association between paranodal axon membrane structure and the functional difference of dorsal and ventral root axons.

Action Potentials↗
PubMed1986

Axons regenerated through silicone tube splices. I. Conduction properties.

Changes in conduction properties of axons regenerating across a 10-mm gap bridged by a silicone cuff were investigated from compound action potential responses. Compound action potentials were detected as early as 6 weeks after surgery, and were small and slowly conducted at maximum velocities of about 3 m/s. With longer regeneration time, the potentials increased in size, velocity, and complexity. Conduction velocities increased rapidly at first than slowly and asymptotically approached rates that were approximately 40% below normal after 10 months. One component of the compound action potential, the refractory period, decreased from 5 ms to near normal value after only 3 months. The time constant of excitation changed more rapidly, and after 2 months approximated values near those for controls. The properties of axons regenerated across an epineural suture with no gaps. The database for the time course of events established here will be useful in guiding studies using the silicone cuff technique as an in situ experimental chamber for studies of regeneration and remyelination.

Animals↗
PubMed1986

Axons regenerated through silicone tube splices. II. Functional morphology.

The recovery of axons regenerated through silicone tube splices was studied with electron microscopic and morphometric methods. Regenerated nerves contained both myelinated and unmyelinated axons of near normal morphology. The number and diameter of axons increased with postoperative time, and size-frequency histograms demonstrated that regeneration occurred in all major fiber groups. Remyelination occurred between about 4 and 6 weeks. Some of the smallest regenerated axons had unusually thick myelin sheaths, but overall regenerated axons had a slightly thinner sheath compared with similar-size normal fibers, although the ratio of sheath thickness to axon size was within the normal limits of g = 0.65 to 0.8 by 6 weeks. Axons did not, however, regain their normal size within 10 months of surgery. This deficit was apparently the primary factor limiting conduction velocity in these regenerated axons.

Animals↗
PubMed1985

Synaptic morphology and differences in sensitivity.

A relation between synaptic morphology and physiology was observed in an in vitro preparation of a sense organ (the ampulla of Lorenzini), in which activity was monitored from the primary afferent neurons before electron microscopic examination of the afferent synapses. The depth of the postsynaptic trough decreased as prefixation sensitivity of the sense organ decreased. This relation and other ultrastructural differences suggest that physiological properties of synapses are influenced by morphological features. Thus, synapses might be morphologically dynamic to modulate synaptic efficacy in relatively long-term phenomena.

Animals↗
PubMed1984

Evolution of myelin sheaths: both lamprey and hagfish lack myelin.

Modern views of agnathan phylogeny consider Petromyzoniformes and Myxiniformes to belong to distinct classes that diverged from a common ancestor at a remote period, perhaps in the lower Cambrian, greater than 600 million years ago. Both are more primitive than elasmobranchs, holocephalans and bony fishes. Myelin is well developed in elasmobranchs and other fishes but was reported to be lacking in the spinal cord of lampreys. In order to search further for possible early myelin in some part of the nervous system of one of the agnathan stems, or for further evidence that it first appeared in chondrichthians, we extended the sampling to many parts of the brain and cord of hagfish. Transmission electron microscopy was used as a nearly ideal criterion. We find no trace or forerunner of the spiral, multilaminate glial wrapping. Many axons are embedded within one or more glial cells, like unmyelinated fibers in other vertebrates, or lie contiguously in bundles without an obviously complete glial investment. True myelin must be presumed to have been invented within the vertebrates, in ancestors of the living cartilaginous fishes after the agnathans branched from the vertebrate stem.

Animals↗
PubMed1983

Growth and plating efficiency of methanococci on agar media.

Plating techniques for cultivation of methanogenic bacteria have been optimized for two members of the genus Methanococcus. Methanococcus maripaludis and Methanococcus voltae were cultivated on aerobically and anaerobically prepared agar media. Maintenance of O(2) levels below 5 mul/liter within an anaerobic glove box was necessary during plating and incubation for 90% recovery of plated cells. Under an atmosphere of H(2), CO(2), and H(2)S (79:20:1), 2 to 3 days of incubation at 37 degrees C were sufficient for the formation of visible colonies. The viability of plated cells was significantly affected by the growth phase of the culture, H(2)S concentration, and the volume of medium per plate. In addition, colony size of methanococci was affected by agar type, as well as by the concentrations of agar, H(2)S, and bicarbonate.

Journal Article↗
PubMed1980

Electroreception in the ratfish (Hydrolagus colliei).

Behavioral and neurophysiological experiments and anatomical work indicate that the ampullar structures on the head of fish of the subclass Holocephali are sense organs responsive to weak electric fields and are functionally and structurally homologous to the ampullae of Lorenzini in elasmobranchs. It is concluded that, as in elasmobranchs, these organs are used to detect bioelectric and other natural electric phenomena in the environment.

Action Potentials↗
PubMedDate not supplied

DNA fingerprinting confirms isogenicity of androgenetically derived rainbow trout lines.

Homozygous and hybrid clonal lines of rainbow trout (Oncorhynchus mykiss) were confirmed to be isogenic using multilocus DNA fingerprinting. Homozygous clones were produced by androgenesis and gynogenesis using gametes from androgenetic male and female rainbow trout, respectively. Isogenic F1 hybrid lines were produced by crossing homozygous fish from different strains. One line of hybrid clones showed segregation for maternally inherited DNA fingerprint markers. The female from this cross, the only presumptive homozygous gynogenetic individual used in this study, was thought to have been produced by gynogenesis followed by blockage of the first cleavage division, but based on the DNA fingerprint analysis, apparently was derived by spontaneous polar body retention that maintained heterozygosity at some loci. Mutations at DNA fingerprint loci were not observed, indicating relative stability of fingerprint loci in the clonal lines. DNA fingerprinting appears to be a useful tool for identifying and genetically monitoring clonal lines of rainbow trout. Isogenic lines of rainbow trout will facilitate the production of saturated genetic maps for rainbow trout and enhance such endeavors as quantitative trait locus (QTL) analysis and loss of heterozygosity (LOH) studies in tumors.

Androgens↗