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R F Stocker

Publications and source records attributed to R F Stocker.

33 records · Page 2Linked to original sources

The development of the sensory neuron pattern in the antennal disc of wild-type and mutant (lz3, ssa) Drosophila melanogaster.

The development of the sensory neuron pattern in the antennal disc of Drosophila melanogaster was studied with a neuron-specific monoclonal antibody (22C10). In the wild type, the earliest neurons become visible 3 h after pupariation, much later than in other imaginal discs. They lie in the center of the disc and correspond to the neurons of the adult aristal sensillum. Their axons join the larval antennal nerve and seem to establish the first connection towards the brain. Later on, three clusters of neurons appear in the periphery of the disc. Two of them most likely give rise to the Johnston's organ in the second antennal segment. Neurons of the olfactory third antennal segment are formed only after eversion of the antennal disc (clusters t1-t3). The adult pattern of antennal neurons is established at about 27% of metamorphosis. In the mutant lozenge3 (lz3), which lacks basiconic antennal sensilla, cluster t3 fails to develop. This indicates that, in the wild type, a homogeneous group of basiconic sensilla is formed by cluster t3. The possible role of the lozenge gene in sensillar determination is discussed. The homeotic mutant spineless-aristapedia (ssa) transforms the arista into a leg-like tarsus. Unlike leg discs, neurons are missing in the larval antennal disc of ssa. However, the first neurons differentiate earlier than in normal antennal discs. Despite these changes, the pattern of afferents in the ectopic tarsus appears leg specific, whereas in the non-transformed antennal segments a normal antennal pattern is formed. This suggests that neither larval leg neurons nor early aristal neurons are essential for the outgrowth of subsequent afferents.

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Neuronal architecture of the antennal lobe in Drosophila melanogaster.

Computer reconstruction of the antennal lobe of Drosophila melanogaster has revealed a total of 35 glomeruli, of which 30 are located in the periphery of the lobe and 5 in its center. Several prominent glomeruli are recognizable by their location, size, and shape; others are identifiable only by their positions relative to prominent glomeruli. No obvious sexual dimorphism of the glomerular architecture was observed. Golgi impregnations revealed: (1) Five of the glomeruli are exclusive targets for ipsilateral antennal input, whereas all others receive afferents from both antennae. Unilateral amputation of the third antennal segment led to a loss of about 1000 fibers in the antennal commissure. Hence, about 5/6 of the approximately 1200 antennal afferents per side have a process that extends into the contralateral lobe. (2) Afferents from maxillary palps (most likely from basiconic sensilla) project into both ipsi- and contralateral antennal lobes, yet their target glomeruli are apparently not the same as those of antennal basiconic sensilla. (3) Afferents in the antennal lobe may also stem from pharyngeal sensilla. (4) The most prominent types of interneurons with arborizations in the antennal lobe are: (i) local interneurons ramifying in the entire lobe, (ii) unilateral relay interneurons that extend from single glomeruli into the calyx and the lateral protocerebrum (LPR), (iii) unilateral interneurons that connect several glomeruli with the LPR only, (iv) bilateral interneurons that link a small number of glomeruli in both antennal lobes with the calyx and LPR, (v) giant bilateral interneurons characterized by extensive ramifications in both antennal lobes and the posterior brain and a cell body situated in the midline of the suboesophageal ganglion, and (vi) a unilateral interneuron with extensive arborization in one antennal lobe and the posterior brain and a process that extends into the thorax. These structural results are discussed in the context of the available functional and behavioral data.

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Fine structure of a sensory organ in the arista of Drosophila melanogaster and some other dipterans.

The arista, a characteristic appendage of dipteran antennae, consists of 2 short segments at the base and a long distal shaft. A small sensory ganglion, from which arises the aristal nerve, is located proximally in the shaft. The fine structure of the aristal sensory organ was studied in detail in the fruitfly (Drosophila) and for comparison in the housefly (Musca) and the blowfly (Calliphora). In Drosophila, the aristal sense organ consists of 3 identical sensilla that terminate in the hemolymph space of the aristal shaft, and not in an external cuticular apparatus. Each sensillum comprises 2 bipolar neurons and 2 sheath cells; a third sheath cell envelops the somata of all six neurons of the ganglion. The neurons have long slender dendrites with the usual subdivision into an inner and an outer segment. One of the outer segments is highly lamellated and bears small particles (BOSS-structures) on the outside of its cell membrane; the other outer segment is unbranched and has a small diameter. The fine structure of the first dendrite is strongly reminiscent of thermoreceptors known from the antennae of other insects. These thermoreceptors are often coupled with hygroreceptors; however, we can only speculate whether the second dendrite of the aristal organ also has this function. Our present results argue against mechanoreceptive functions, as formerly postulated. The aristal sense organs in Musca and Calliphora are similar to those in Drosophila, but contain more sensilla (12 in Musca, 18 in Calliphora.

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Courtship behavior of Drosophila genetically or surgically deprived of basiconic sensilla.

The lack of basiconic antennal sensilla in the mutant lozenge was used to assess the role of these olfactory receptors in the courtship behavior of Drosophila melanogaster. Under normal light conditions, lozenge males courted virgin females much less than wild-type males did. However, when visual courtship stimuli were eliminated by studying behavior under dim red light, the two kinds of males courted individual wild-type virgin females with the same intensity, and the latency to copulation was similar. Also, no difference in courtship vigor was observed if the two kinds of males were paired in red light with a mated female. These data suggest that antennal basiconic sensilla are important for neither the perception of the attraction pheromone(s) of virgin females nor the inhibitory pheromone(s) of mated females. Similar assays with males deprived of maxillary palps make it unlikely that the basiconic-like sensilla on these appendages are needed to perceive the attraction pheromones. However, the unexpectedly high courtship activity of palp-deprived males toward mated females suggests that basiconic-like maxillary sensilla may be receptors of inhibitory female compounds.

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Peripheral and central nervous effects of lozenge3: a Drosophila mutant lacking basiconic antennal sensilla.

The third antennal segment (= funiculus) of wild-type Drosophila melanogaster shows a sexually dimorphic distribution of sensilla: Males possess about 20% less of large basiconic sensilla, but approx 30% more trichoid sensilla than the female. The funiculus of the mutant lozenge3 is much reduced in size. Moreover, basiconic sensilla are completely lacking, and the number and density of trichoid sensilla are reduced. In contrast, the number and density of coeloconic sensilla are increased. The loss of sensilla in lozenge3 leads to a corresponding loss of sensory fibers in the antennal nerve. The antennal commissure of the wild type consists essentially of afferents from the funiculus which extend into the contralateral half of the brain. In the antennal commissure of lozenge3, more than twice the number of fibers lacking in the antennal nerve have disappeared which suggests that most afferents establish purely ipsilateral terminals. A highly specific change in the brain of lozenge3 is the loss of a particular subunit of the antennal center, the glomerulus V. This has previously been shown to be a major target of fibers from basiconic sensilla. Mosaic flies exhibiting a lozenge3 antenna demonstrate that the elimination of glomerulus V is causally related to the change in the sensilla pattern. This implies that the development and/or survival of particular target regions in the antennal center depends on sensory input. Furthermore, it shows that glomerulus V is specifically involved in the processing of information from basiconic sensilla.

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Projection patterns of different types of antennal sensilla in the antennal glomeruli of Drosophila melanogaster.

Cobalt fills from small, defined regions of the antenna in D. melanogaster show that the three types of sensilla on the third segment, the flagellum, and a fourth sensillum located in the arista, project into the glomeruli of the antennal lobe. We have identified 19 glomeruli in each lobe, according to their location, shape, and size. At least ten of these represent major projection areas of flagellar or aristal sensilla. The large majority of glomeruli is innervated from both antennae, but a small group of five receive exclusively ipsilateral input. A particular sensory fiber appears to terminate only in one specific glomerulus, either in the ipsilateral or in both lobes. Fills from flagellar regions bearing a single type of sensillum, yield a specific pattern of glomeruli containing stained terminals. Aristal projections remain strictly ipsilateral, whereas those from the other sensilla consist of an ipsilateral and a bilateral component. When filling from different points in an area bearing one type of sensillum, similar projections are produced, suggesting that projection patterns observed reflect predominantly the type of sensillum rather than its location on the flagellum. Accordingly, individual glomeruli might represent functional units, each receiving antennal input in a characteristic combination.

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Cobalt filling of sensory projections from internal and external mouthparts in Drosophila.

Phase-contrast microscopy of 2.5 micron plastic sections reveals five different kinds of sense organs on the internal mouthparts of Drosophila melanogaster: a ventral group of sensilla on the epipharynx, three kinds of sense organs in the middle part of the sucking apparatus (the cibarium), one of them being described here for the first time, and an additional type of sensillum in the dorsal pharynx (Fig. 1). Three of these internal sense organs resembling each other structurally form similar projection patterns in the tritocerebrum, whereas external taste receptors on the labial palps show a different pattern in the suboesophageal ganglion (Fig. 12). The central projections of these internal and external sensilla were traced by applying an ethanolic solution of cobalt on the intact cuticle.

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Fine structural comparison of the antennal nerve in the homeotic mutant Antennapedia with the wild-type antennal and second leg nerves of Drosophila melanogaster.

In D. melanogaster the cross-sectioned nerve of the leg-like antenna in the homeotic mutant Antennapedia was ultrastructurally compared with the nerves of the morphologically related second leg and the wild-type antenna. The nerves of the normal antenna and the second leg differ from one another in both the numbers and arrangement of axons. According to these criteria the nerve of the homeotic appendage was structurally identified as a leg nerve. Most of the antennal nerves studied showed a consistent grouping of axons in the profile. This suggests that the assemblage of the axons does not occur randomly, but in an ordered fashion.

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Fine structure of degenerating abdominal motor neurons after eclosion in the sphingid moth, Manduca sexta.

Ultrastructural aspects of the natural degeneration of a group of six motor neurons in the fourth abdominal ganglion of Manduca sexta are described. These motor neurons innervate intersegmental muscles that degenerate and disappear immediately after adult eclosion. The first detectable changes in the cell bodies appear 12h after eclosion and include disruption of the endoplasmic reticulum and an increase in the size and number of lamellar bodies. At 32h the nuclear membranes rupture, and the membranous and granular cytoorganelles segregate in different parts of the cell. At that stage the surrounding glial cells participate in the digestion of material from the degenerating neurons. From 72h onward the remaining neuronal structures become disrupted, and are finally transformed into a single, large lamellar body (residual body) within the glial profile. The degeneration pattern differs significantly from that of embryonic vertebrate neurons.

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Projection of sensory neurons from a homeotic mutant appendage, Antennapedia, in Drosophila melanogaster.

Central projections of sensory neurons from homeotic mutant appendages (Antennapedia) of Drosophila melanogaster were compared with those of wild-type antennae and wild-type legs by means of degeneration and cobalt backfilling methods. Sensory axons originating from wild-type thoracic legs terminate within the ventral ipsilateral half of the corresponding neuropile segment and do not project to the brain. Sensory fibers from the third antennal segment (AIII) of wild-type animals project into the ipsilateral antennal glomerulus (AG) and to a lesser extent into the contralateral AG, whereas those from the second antennal segment terminate principally within the ipsilateral posterior antennal center. The sensory terminals of femur, tibia, and tarsi of the homeotic leg show a distribution very similar to that of the homologous wild-type antennal segment AIII, differing to a minor degree only in the size and precise localization of terminals within the antennal glomeruli. No degenerating axons were evident in ultrastructural examination of neck connectives after removal of homeotic legs. It is thus very improbable that any sensory fibers of the homeotic leg project to normal leg projection areas in the thoracico-abdominal ganglion. Several alternative explanations are offered for the apparent retention of antennal specificity by axons from the transformed appendage.

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Ultrastructural studies on neuromuscular contacts and the formation of junctions in the flight muscle of Antheraea polyphemus (Lep.). II. Changes after motor nerve section.

In the moth Antheraea polyphemed at the onset of adult development. The subsequent breakdown of the isolated motor stulongated vesicles similar in structure to channels of smooth ER, appear in large numbers in the axoplasm. Their nature as well as the functional aspects of early axonal changes are discussed. From the 7th day onward two types of axonal breakdown become prominent. The first is characterized 0y swelling axon profiles, distorted vesicles and strongly shrunken mitochondria, uhile shrinking axon profiles containing tightly packed mitochondria and unaltered vesicles are typical of the second. Both types presumably take place independently of each other in different axon terminals. Axons and the contents of at least the first type are finally removed by transformation into lamellar bodies. Glial processes obviously behave independently of degenerating terminals; they loose any contact with them and never act as phagocytes for axon remnants. During the whole period of breakdown undifferentiated contacts between nerve fibers and muscle anlagen are present but synaptic structures as in normal developing dlm have never been observed. This fact, in comparison with earlier studies, suggests a lack of trophic nervous activity on the muscle anlagen tissue. A short time after removal of the isolated stumps new nerve tracts appear between dlm-fibers (which are, of course, strongly retarded in development). They are presumably sensory wing nerves which lack a guide structure to the central target, due to axotomy. Neuromuscular contacts or even junctions formed by axons of these nerves have occasionally been detected on the dlm. Their nature is discussed. Wallerian axon degeneration is compared to the normal, metamorphic breakdown of the innervation of the larval dlm-precursor. In contrast to the former, glial processes here remain in contact with the terminals. Glia and axons first swell. Then most glial processes are transformed into lamellar bodies whereas neurites shrink and become electron-dense. Axonal organelles remain intact for a long period.

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Ultrastructural studies on neuromuscular contacts and the formation of junctions in the flight muscle of Antheraea polyphemus (Lep.) I. Normal adult development.

The ultrastructure of neuromuscular connections on developing dorsolongitudinal flight muscles were studied in the moth Antheraea polyphemus. Undifferentiated membrane contacts between axon terminals and muscle-fiber anlagen are present in the diapause pupa. They persist during the period of nerve outgrowth, which probably provides a pathway of contact guidance. By the 4th day of adult development some of these contact areas have differentiated into structures similar to neuromuscular junctions although differentiation of muscle structure does not start earlier than the eighth day. Dense-cored vesicles are abundant in many axon terminals at the beginning of development. They later decrease in number quite rapidly. The significance of the above-mentioned early junctions, their possible mode of action and the role of the dense-cored vesicles are discussed. It is proposed that they exercise a stimulating (trophic) influence on the growth of the undifferentiated muscular tissue. The imaginal neuromuscular junctions are formed during the second half of adult development. Clusters of vesicles and electron-dense depositions along the inner face of the axo- and lemma seem to initiate junction formation. Glial processes then grow between the axo- and sarcolemma and divide the large contact area into several small segments. Mutual invaginations and protrusions of the sarcolemma and the glial cell membrane subsequently form an extensive "rete synapticum." Six days before eclosion the glial and sarcoplasmic parts of the rete synapticum are similar in size. Up to eclosion, all glial processes shrink and increase in electron density. Most of the observations are discussed also in relation to findings in vertebrates.

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