PubMed Health⌕ Search

Biomedical subjects

T S Reese

Publications and source records attributed to T S Reese.

At least 91 records · Page 5Linked to original sources
PubMed1985

Structure of axon terminals and active zones at synapses on lizard twitch and tonic muscle fibers.

The freeze-fracture technique was used to study differences in membrane structure which could explain differences in the number of quanta released from axon terminals on twitch and tonic muscle fibers in Anolis intercostal muscles. The protoplasmic leaflets of axon terminals facing lizard twitch muscle fibers have intramembrane particle specializations characterized by two parallel linear particle arrays each composed of two particle rows which lie perpendicular to the axis of shallow ridges in the axolemma. During K+ depolarization, vesicles open between the arrays, confirming that these structures are the active zones for synaptic vesicle opening. Active zones at axon terminals on tonic fibers are defined by one linear particle array composed of two parallel particle rows oriented along the axis of a shallow presynaptic ridge; vesicles open beside these arrays. Thus, there are more particles near active zone vesicles in terminals on twitch fibers. Even though terminals on twitch and tonic muscle fibers seem to have similar numbers of synaptic vesicles associated with their active zones, a presynaptic action potential is reported to release at least 10 times more quanta from terminals on twitch fibers. We postulate that the differences in quantal output are related to the observed differences in the number of active zone particles flanking synaptic vesicles at the active zone. Indeed, the correlation between the distribution of these particles and the level of transmitter release provides additional support for the idea that they are the calcium channels which couple transmitter release to the action potential.

Animals↗
PubMed1984

Structure and distribution of neuromuscular junctions on slow muscle fibers in the frog.

Neuromuscular junctions on slow fibers in frog cruralis muscle have been examined with light and electron microscopy. The slow fiber bundle of this muscle is approximately half slow fibers and half fast twitch fibers. Each slow fiber has two-five clusters of nerve terminals in its central region. In contrast, fast fibers in the same bundle have single anastomosing neuromuscular junctions. The average length of terminal clusters on slow fibers is half that of the terminals on fast fibers. Less cholinesterase activity is associated with nerve terminals on slow muscle fibers. The two types of muscle fiber were identified in freeze-fracture replicas by characteristic patterns of the sarcolemmal caveolae and square arrays. Presynaptic membranes of terminals on fast fibers have long, paired double rows of intramembrane particles which lie along the sides of ridges aligned above each fold in the muscle. Ridges are less prominent or absent in terminals on slow fibers and the associated membrane particles are more often in single rows. The lengths of the particle rows also tend to be shorter and rows tend to branch and to lie at various orientations with respect to the longitudinal axis of the terminal. The average length of the particle rows per unit length of nerve terminal on slow fiber terminals is half that of particle rows on fast fiber terminals. It is concluded that the total length as well as the concentration of these active zone specializations is less at terminals on slow fibers. The lack of parallel orientation of the presynaptic active zones at slow fiber terminals corresponds to a lack of postsynaptic folds. Postsynaptic specializations seen in thin sections through slow fibers are either on flat expanses of sarcolemma or on low sarcolemmal bulges. Patches of large intramembrane particles, similar to those on fast fibers, mark regions of postsynaptic specialization in freeze-fracture replicas of slow fibers.

Animals↗
PubMed1984

The structure of cytoplasm in directly frozen cultured cells. I. Filamentous meshworks and the cytoplasmic ground substance.

Cultured fibroblasts or epithelial cells derived from Xenopus laevis embryos were directly frozen, freeze-substituted by an improved method, and then either critical-point-dried and viewed as whole mounts, or embedded and thin sectioned. In thin regions of these cells, where ice crystal artifacts are absent, the cytoplasm consisted of a dense, highly interconnected meshwork of filaments, embedded in a finely granular ground substance. The meshwork in directly frozen, intact cells was compared with that in cells that were lysed (physically, with detergents, or with filipin), or fixed with glutaraldehyde before freezing. Although filaments tended to be less numerous in lysed cells, their overall organization was the same as that in intact cells. However, fixation with glutaraldehyde before freezing distorted the meshwork to variable degrees depending on the osmolarity of the fixation buffer, and also obscured the granular ground substance which is obvious in directly frozen cells. With optimal preparative methods, the cytoplasm of these directly frozen cells is shown to consist of a cytoskeleton composed of discrete interwoven filaments interconnected by numerous finer filaments and a readily extractable granular matrix which presumably represents aggregations of cytoplasmic proteins.

Animals↗
PubMed1983

Inhibitory and excitatory synapses in crayfish stretch receptor organs studied with direct rapid-freezing and freeze-substitution.

Crayfish abdominal stretch receptor organs are innervated by inhibitory (GABA) and excitatory (glutamate) synapses. Previous studies with aldehyde fixation showed that synaptic vesicles in the inhibitory synapse are flat and small, whereas those in the excitatory synapse are rounder and larger. We have reexamined these inhibitory and excitatory synapses by using direct rapid-freezing and freeze-substitution in order to preserve synaptic structure closer to its living state. Fine details of synaptic structure appear to be better preserved by this method. Synaptic vesicles in inhibitory as well as excitatory synapses are round, so the conventional flattened shape of vesicles in the inhibitory synapse must depend on some aspect of aldehyde processing. However, the average size of vesicles in the inhibitory synapse is significantly smaller than that of vesicles in the excitatory synapse, so synaptic vesicle size is regarded as having functional significance.

Animals↗
PubMed1983

Formation of misplaced and reflexive tight junction strands in prostate epithelial cells.

Tight junction strands occur at three atypical locations in slices of rat ventral prostate exposed to conditions promoting rapid tight junction assembly: (1) in the basal plasma membranes of the columnar epithelial cells, over 40 microns from the native apical tight junction band (misplaced tight junctions); (2) in the plasma membranes of basal epithelial cells, which never have tight junctions in the native state; and (3) between processes of the same cell (reflexive or autocellular tight junctions) at the basal or lateral portions of the columnar epithelial cell. These findings suggest that tight junction formation is not limited to specific parts of the plasma membrane, even in highly polarized cells such as those in prostate epithelium. Taken together with other new evidence, they also suggest that tight junctions may be very labile.

Animals↗
PubMed1983

Cytoplasmic organization in cerebellar dendritic spines.

Three sets of filamentous structures were found to be associated with synaptic junctions in slices of cerebellar tissue prepared by rapid-freezing and freeze-etch techniques. The electron-dense fuzz subjacent to postsynaptic membranes corresponds to a web of 4-6-nm-diam filaments that were clearly visualized in rapid-frozen, freeze-etched preparations. Purkinje cell dendritic spines are filled with a meshwork of 5-7-nm filaments that were found to contact the spine membrane everywhere except at the synaptic junction, and extend through the neck of the spine into the parent dendrite. In addition, 8-10-nm microfilaments, possibly actin, were seen to be associated with the postsynaptic web and to extend into the body and neck of the spine. The arrangements and attachments of the filamentous elements in the Purkinje cell dendritic spine may account for its shape.

Animals↗
PubMed1983

Structural changes during transmitter release at synapses in the frog sympathetic ganglion.

Interactions between synaptic vesicles and the presynaptic membrane which accompany transmitter release were examined at excitatory, cholinergic synapses in bullfrog sympathetic ganglia. Ganglia were fixed at rest or during electrical stimulation of the preganglionic axons and then were either thin-sectioned or freeze-fractured. Release of transmitter for brief periods is accompanied by selective depletion of four-fifths of the synaptic vesicles aligned at active zones, an overall loss of half of the synaptic vesicles in the terminals, and synaptic vesicle openings at active zones. These findings are consistent with the hypotheses that synaptic vesicles which are ready to be released are aligned at active zones and that these vesicles fuse with and add their membrane to the presynaptic membrane as they release transmitter. Larger vesicles with dense cores also contact and open onto the presynaptic membrane at the active zone, appearing to release their contents by exocytosis. The arrangement of intramembrane particles at fractured postsynaptic specializations resembles that at other excitatory, cholinergic synapses.

Animals↗
PubMed1982

Regional organization of astrocytic membranes in cerebellar cortex.

The internal organization of plasmalemmal membrane, as revealed by freeze-fracture techniques, varies dramatically and predictably over the surface of astrocytes in mouse cerebellar cortex. Assemblies of uniform, small intramembrane particles packed in orthogonal order into square or rectangular aggregates are specialized distribution of intramembrane particles which, in the cerebellar cortex, are found only in astrocytes. The concentration of assemblies is greatest in astrocytic membrane juxtaposed to vascular structures or facing the cerebrospinal fluid at the glial limitans. Many fewer are present in regions of astrocytic membrane apposed to neural structures and virtually none are present on the astrocyte cell body. Corresponding structures have not yet been found in thin-sectioned preparations. While the distribution of assemblies in membranes facing blood and cerebrospinal fluid compartments suggests that they may have a role in transport of some material into or out of those compartments, their function is unknown. A second, distinct specialization of intramembrane structure appears to represent a junction between apposed astrocytic processes. We have provisionally described this as a 'polygonal particle junction', since it appears as large, irregular particles densely packed without obvious order in co-extensive regions of two astrocytic membranes. This junction is regularly present just below the cerebellar surface in the processes of the glial limitans as well as between large, more proximal radial Bergmann fibers, and also occurs occasionally throughout the molecular layer. With tannic acid mordant after aldehyde-osmium fixation or rapid freezing and freeze-substitution, it is possible to demonstrate subtle electron-dense specializations of the astrocytic membranes and extracellular matrix in thin-sections which correspond to the sites of polygonal particle junctions. The function of this astrocytic specialization is also unknown. Cerebellar astrocytes manifest numerous gap junctions as well, whose structure in freeze-fractured and thin-sectioned preparations is quite distinct from that of assemblies or of polygonal particle junctions.

Animals↗
PubMed1982

Cytoplasmic structure in rapid-frozen axons.

Turtle optic nerves were rapid-frozen from the living state, fractured, etched, and rotary shadowed. Stereo views of fractured axons show that axoplasm consists of three types of longitudinally oriented domains. One type consists of neurofilament bundles in which individual filaments are interconnected by a cross-bridging network. Contiguous to neurofilament domains are domains containing microtubules suspended in a loose, granular matrix. A third domain is confined to a zone, 80-100 nm wide, next to the axonal membrane and consists of a dense filamentous network connecting the longitudinal elements of the axonal cytoskeleton to particles on the inner surface of the axolemma. Three classes of membrane-limited organelles are distinguished: axoplasmic reticulum, mitochondria, and discrete vesicular organelles. The vesicular organelles must include lysosomes, multivesicular bodies, and vesicles which are retrogradely transported in axons, though some vesicular organelles may be components of the axoplasmic reticulum. Organelles in each class have a characteristic relationship to the axonal cytoskeleton. The axoplasmic reticulum enters all three domains of axoplasm, but mitochondria and vesicular organelles are excluded from the neurofilament bundles, a distribution confirmed in thin sections of cryoembedded axons. Vesicular organelles differ from mitochondria in at least three ways with respect to their relationships to adjacent axoplasm: (a) one, or sometimes both, of their ends are associated with a gap in the surrounding granular axoplasm; (b) an appendage is typically associated with one of their ends; and (c) they are not attached or closely apposed to microtubules. Mitochondria, on the other hand, are only rarely associated with gaps in the axoplasm, do not have an appendage, and are virtually always attached to one or more microtubules by an irregular array of side-arms. We propose that the longitudinally oriented microtubule domains are channels within which organelles are transported. We also propose that the granular material in these channels may constitute the myriad enzymes and other nonfibrous components that slowly move down the axon.

Animals↗
PubMed1981

A freeze-fracture study of photoreceptor presynaptic membranes during ribbon synapse formation.

The outer plexiform layer (OPL) of the developing chick retina from 11 embryonic days to 11/2 weeks posthatching has been studied by freeze-fracture to characterize changes in the membrane structure of photoreceptor terminals during synaptogenesis. At early stages, the undifferentiated photoreceptor synaptic base is characterized by a sparse distribution of intramembrane articles on the inner leaflet (P-face). Later, as the synaptic base begins to differentiate by extending filopodia into the OPL, numerous small aggregates of large particles appear between and on filopodial surfaces. Many of the aggregates occupy crater-like expressions, which are seen in cross-fractures through the underlying cytoplasm to be associated with vesicular invaginations of the presynaptic membrane. Corresponding thin sections through these regions at this time reveal immature arciform densities and coated vesicles fusing with the presynaptic membrane adjacent to these densities. At later stages, many of the particle aggregates on the photoreceptor membrane appear to have coalesced into longer arrays overlying ridges surrounded by numerous vesicle fusion sites. These intramembrane changes correlate with the formation of the mature arciform density-synaptic ribbon specialization in the photoreceptor presynaptic terminal and with physiological maturation of the chick retina.

Animals↗
PubMed1981

Are the presynaptic membrane particles the calcium channels?

The number of large intramembrane particles associated with sites of synaptic vesicle release at the squid giant synapse was determined and compared to the average maximal presynaptic calcium current in order to derive an estimate of the conductance each particle would have if it were a calcium channel. This value, 0.21 pS, compares favorably with conductances of calcium channels in other preparations, substantiating the idea that the large intramembrane particles, which are concentrated at "active zones," represent calcium channels.

Animals↗
PubMed1981

Structural changes after transmitter release at the frog neuromuscular junction.

The sequence of structural changes that occur during synaptic vesicle exocytosis was studied by quick-freezing muscles at different intervals after stimulating their nerves, in the presence of 4-aminopyridine to increase the number of transmitter quanta released by each stimulus. Vesicle openings began to appear at the active zones of the intramuscular nerves within 3-4 ms after a single stimulus. The concentration of these openings peaked at 5-6 ms, and then declined to zero 50-100 ms late. At the later times, vesicle openings tended to be larger. Left behind at the active zones, after the vesicle openings disappeared, were clusters of large intramembrane particles. The larger particles in these clusters were the same size as intramembrane particles in undischarged vesicles, and were slightly larger than the particles which form the rows delineating active zones. Because previous tracer work had shown that new vesicles do not pinch off from the plasma membrane at these early times, we concluded that the particle clusters originate from membranes of discharged vesicles which collapse into the plasmalemma after exocytosis. The rate of vesicle collapse appeared to be variable because different stages occurred simultaneously at most times after stimulation; this asynchrony was taken to indicate that the collapse of each exocytotic vesicle is slowed by previous nearby collapses. The ultimate fate of synaptic vesicle membrane after collapse appeared to be coalescence with the plasma membrane, as the clusters of particles gradually dispersed into surrounding areas during the first second after a stimulus. The membrane retrieval and recycling that reverse this exocytotic sequence have a slower onset, as has been described in previous reports.

Animals↗
PubMed1981

Astrocyte membrane structure: changes after circulatory arrest.

Membranes of the astrocytic processes investing small blood vessels and the surface of the brain contain numerous arrays of orthogonally packed particles as revealed by the freeze-fracture technique. The structure of these particle arrays, which we have termed "assemblies," is the same whether tissue is prepared for freeze-fracture by conventional fixation or by quick excision and rapid freezing. However, assemblies are progressively replaced by amorphous clumps and then disappear as the interval between decapitation and rapid freezing increases. Nearly normal numbers of assemblies may be maintained in cerebellar slices in vitro, but there too they disappear at low PO2 or in the presence of dinitrophenol. No other neuronal or glial membrane specialization exhibits a comparable lability.

Animals↗
PubMed1981

Beginning of exocytosis captured by rapid-freezing of Limulus amebocytes.

Structural changes underlying exocytosis evoked by the application of endotoxin to Limulus amebocytes were studied at the level of detail afforded by freeze-fracture and freeze-substitution techniques combined with the time resolution of direct rapid-freezing. The results with amebocytes prepared in this manner differed from those with other secretory cells prepared by conventional means. Exocytosis begins within seconds of endotoxin treatment when the plasmalemma invaginates to form pedestallike appositions with peripheral secretory granules. The juxtaposed membranes at these pedestal appositions form several punctate pentalaminar contacts, but examination of freeze-fractured pedestals failed to reveal any corresponding changes in the intramembrane particle distribution. Small secretory granule openings or pores, which are very infrequent, appear within the first 5 s after endotoxin treatment. These pores rapidly widen and this widening is immediately followed by the sequential dissolution of the granule contents, which then move into the surrounding extracellular space. Cytoplasmic filaments connecting the plasmalemma with the granule membrane are suitably deployed to be responsible for the plasmalemma invaginations. How pores begin is not certain, but the appearance of clear spaces between the granule core and the granule membrane at this point in exocytosis supports the possibility of a role of osmotic forces.

Animals↗