Organization and synaptic ultrastructure of glomeruli in the antennal lobes of the moth Manduca sexta : a study using thin sections and freeze-fracture

The antennal lobe of the brain of Manduca sexta comprises a central area of coarse neuropil surrounded by dense, spheroidal glomeruli, where all synaptic interactions between antennal-nerve axons and the second-order neurons of the lobe occur. Neuronal interactions in the glomeruli are complex, involving several types of neuritic profiles and mediated by synapses with a one-to-many ratio of pre- to postsynaptic elements. Presynaptic profiles in the glomeruli have been categorized into three types, containing round clear vesicles, large numbers of large dense-cored vesicles, and pleiomorphic clear vesicles, respectively. Preliminary studies of horseradish peroxidase-filled axons and neurons indicate that antennal-nerve axons form synapses without large numbers of dense-cored vesicles and that antennal-lobe neurons not only receive synapses but also may synapse onto other elements in the antennal lobe. A typical synaptic contact involves multiple postsynaptic elements apposed in pairs to an individual presynaptic element. The presynaptic element contains a bar-shaped membrane-associated density, which follows a shallow groove in the membrane and is flanked by synaptic vesicles. Postsynaptic elements are lined by membrane-associated densities in the region opposite to the synaptic bar, and may be observed to participate in serial synapses. Freeze-fracture replicas of the glomerular neuropil contain many membrane specializations that are thought to be presynaptic, some of which resemble those of vertebrate excitatory synapses. At these apparently presynaptic regions, large particles cluster in the P face of the membrane and are often surrounded by plasmalemmal deformations presumably representing sites of exo- or endocytosis. The shape of the predominant type of presynaptic membrane specialization (a plaque) does not match the shape of the presynaptic membrane-associated density (a bar); this raises the possibility that vesicle release occurs at isolated ‘active zones’ along the presynaptic bar. Postsynaptic sites are represented by clusters of large particles in the E face of the postsynaptic membrane.

Author(s):  
Ji-da Dai ◽  
M. Joseph Costello ◽  
Lawrence I. Gilbert

Insect molting and metamorphosis are elicited by a class of polyhydroxylated steroids, ecdysteroids, that originate in the prothoracic glands (PGs). Prothoracicotropic hormone stimulation of steroidogenesis by the PGs at the cellular level involves both calcium and cAMP. Cell-to-cell communication mediated by gap junctions may play a key role in regulating signal transduction by controlling the transmission of small molecules and ions between adjacent cells. This is the first report of gap junctions in the PGs, the evidence obtained by means of SEM, thin sections and freeze-fracture replicas.


1995 ◽  
Vol 198 (3) ◽  
pp. 603-611 ◽  
Author(s):  
P Kloppenburg ◽  
J G Hildebrand

Using intracellular recording techniques, we have begun to examine the effects of 5-hydroxytryptamine (5-HT) on antennal-lobe (AL) neurones in the brain of the adult moth Manduca sexta. 5-HT modulated the responses of local interneurones and projection neurones, which were recognized on the basis of well-established electrophysiological criteria, to primary synaptic input elicited by electrical stimulation of the ipsilateral antennal nerve. 5-HT applied at low concentration (10(-8) mol l-1) reduced the excitatory responses evoked by electrical stimulation of the antennal nerve, whereas at high concentration (10(-4) mol l-1), 5-HT enhanced the responses. At 10(-4) mol l1, 5-HT increased cell input resistance, led to broadening of action potentials and caused increased cell excitability in many AL neurones.


1980 ◽  
Vol 85 (2) ◽  
pp. 337-345 ◽  
Author(s):  
R Fesce ◽  
F Grohovaz ◽  
W P Hurlbut ◽  
B Ceccarelli

The intramembrane particles on the presynaptic membrane and on the membrane of synaptic vesicles were studied at freeze-fractured neuromuscular junctions of the frog. The particles on the P face of the presynaptic membrane belong to two major classes: small particles with diameters less than 9 nm and large particles with diameters between 9 and 13 nm. In addition, there were a few extralarge particles with diameters greater than 13 nm. Indirect stimulation of the muscle, or the application of black widow spider venom, decreased the concentration of small particles on the presynaptic membrane but did not change the concentration of large particles. Three similar classes of particles were found on the P face of the membrane of the synaptic vesicles. The concentrations of large and extralarge particles on the vesicle membrane were comparable to the concentrations of these particles on the presynaptic membrane, whereas the concentration of small particles on the vesicle membrane was less than than the concentration of small particles on the presynaptic membrane. These results are compatible with the idea that synaptic vesicles fuse with the presynaptic membrane when quanta of transmitter are released. However, neither the large nor the extralarge particles on the P face of the presynaptic membrane can be used to trace the movement of vesicle membrane that has been incorporated into the axolemma.


1983 ◽  
Vol 60 (1) ◽  
pp. 289-301
Author(s):  
N.E. Flower ◽  
J.D. Briers

Freeze-fracture studies of Lepidopteran antennae have revealed the presence of orthogonal arrays of particles on antennal nerve membranes. Initial impressions were that several different arrays were present. Optical diffraction was used to examine the arrays on the electron micrograph negatives. This technique showed that all the arrays were derived from the same basic structure, suggesting that the superficial differences in appearance were due to shadowing effects. The particles are arranged in a lattice with spacings of 10.7 nm X 9.1 nm. The arrays are not clear-cut but tend to break up, producing a disorganized region around their edges. The particles are shown to have depressions in them. However, the evidence available suggests that the arrays do not have the other characteristics of gap junctions. The arrays appear not to be present on most of the nerve membrane faces, occurring only in localized regions of the nerve membranes where they are present in large numbers. This suggests that the arrays may have a specialized local function.


1980 ◽  
Vol 28 (2) ◽  
pp. 142-148 ◽  
Author(s):  
K R Fry ◽  
A W Spira

Ethanolic phosphotungstic acid (EPTA) has been used to elucidate the structure of certain organelles contained within retinal cells not clearly discernible using conventional preparations. Both synaptic and nonsynaptic components of the guinea pig neural retina have been analyzed. Within the photoreceptor (PR) cell EPTA-stained components include the connecting cilia, their basal bodies, and the root filament system. Cross-striated fibrillar organelles, similar in appearance to the root filaments, are also observed in the nuclear region, the synaptic terminal and other parts of the PR cell. The possible structural continuity and significance of these structures are discussed. Within retinal synapses of both the inner and outer plexiform layers, ribbons and associated paramembranous specializations are stained. The photoreceptor ribbons have a trialaminar structure with filamentous, tufted borders. Synaptic cleft material and postsynaptic densities are also stained. Bipolar cell synapses in the inner plexiform layer contain stained short ribbons as well as closely associated peg-like densities extending towards the presynaptic membrane.


1978 ◽  
Vol 30 (1) ◽  
pp. 151-174
Author(s):  
J.G. Robertson ◽  
M.P. Warburton ◽  
P. Lyttleton ◽  
A.M. Fordyce ◽  
S. Bullivant

Peribacteroid membranes and bacteroid envelope inner membranes have been isolated from developing lupin nodules. Isolation of the peribacteroid membranes was achieved by first preparing membrane-enclosed bacteroids free from other plant organelles or membranes. The peribacteroid membranes were then released by osmotic shock and purified by centrifugation to equilibrium on sucrose gradients. The bacteroids were broken in a pressure cell and the bacteroid envelope inner membranes were isolated using sucrose gradient fractionation of the bacteroid total envelope preparation. The density of the peribacteroid membranes decreased during the period of development of N2-fixation in lupin nodules from 1.148 g/ml for nodules from 12-day plants to 1.137 g/ml for nodules from 18-day plants. The density of the bacteroid envelope inner membranes from nodules from 18-day plants was 1–153 g/ml. The identity and homogeneity of the isolated membranes was established, by comparison with membranes in intact nodules, using phosphotungstic acid and silver staining of thin sections and particle densitites on faces of freeze-fracture replicas of the membranes. Analyses for NADH oxidase and succinate dehydrogenase, spectral analyses and gel-electrophoretic analysis of proteins were also used to characterize the membrane and soluble protein fractions from the nodules. The ratio of lipid to protein was 6.1 for the peribacteroid membranes and 2.5 for the bacteroid envelope inner membranes. Leghaemoglobin was localized in the plant cytoplasm in lupin nodules and not in the peribacteroid space.


1977 ◽  
Vol 27 (1) ◽  
pp. 255-272
Author(s):  
D.S. Smith ◽  
U. Jarlfors ◽  
M.L. Cayer

The distribution of microtubules and mitochondria in central axons of an insect (Periplaneta americana) is assessed by comparison between counts on micrographs and computed axon random ‘models’. These studies show that the observed multiple association of microtubules with individual mitochondria is statistically highly significant. Electron micrographs of thin sections show that linkage is effected by physical cross-bridge, possibly comprising components from the microtubule and mitochondrion. Linear particle arrays are described on the outer mitochondrial membrane in freeze-fracture replicas, and tentatively related to the bridges seen in thin sections. The results are discussed in terms of proposed roles of microtubules in neurons and other cells.


2003 ◽  
Vol 189 (4) ◽  
pp. 301-308 ◽  
Author(s):  
B. S. Hansson ◽  
M. A. Carlsson ◽  
B. Kalinovà

1982 ◽  
Vol 56 (1) ◽  
pp. 245-262 ◽  
Author(s):  
N.J. Lane ◽  
L.S. Swales

The stages that occur during the assembly of both pleated and smooth septate junctions in developing insect tissues have been examined. The oesophagus and mid-gut of the embryonic moth, and the oesophagus and central nervous system (CNS) of the locust embryo, have been investigated in thin sections and by freeze-fracture during the course of membrane biogenesis. The smooth septate junctions developing between the lateral borders of the mid-gut exhibit, in the early stages, individual intramembranous particles becoming aligned into short ridges. These ultimately migrate over the membrane face and fuse into longer arrays, which become stacked in parallel with other ridges to form the characteristic mature form of the junction just before hatching. Pleated septate junctions occur between the cells both of the oesophagus and of the perineurium, which ensheathes the neurones and the neuroglial cells in the locust CNS; these are also fully formed by the end of embryonic development. The pleated junctions appear to be assembled during the later stages of CNS or gut differentiation, arising first in embryos about two-thirds of the way through development. During their maturation, the initial event seems to be a membrane depression in the P face, which occurs in patches over the presumptive junctional membrane. Into these depressed regions or ‘formation-plaque’ areas, 8–10 nm particles appear to be inserted intramembranously in apparently random arrays. These particles are the most common elements but larger particles are also present; the former ultimately become aligned in a row. With time, other intramembranous particles come to lie in rows parallel to the original one. By hatching, the typical undulating stacks of parallel intramembranous particle rows are fully formed. Gap junctions also form between the same perineurial or oesophageal cells, usually before, but in some cases at the same time, or just after, the septate junctions have been assembled. Tricellular associations between cells also appear around the same time in embryonic development. The simultaneous assembly of these different junctions reflects a high degree of organizational capacity at the membrane level.


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