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C Consoulas

Publications and source records attributed to C Consoulas.

12 recordsLinked to original sources
PubMed2004

Analysis of conditional paralytic mutants in Drosophila sarco-endoplasmic reticulum calcium ATPase reveals novel mechanisms for regulating membrane excitability.

Individual contributions made by different calcium release and sequestration mechanisms to various aspects of excitable cell physiology are incompletely understood. SERCA, a sarco-endoplasmic reticulum calcium ATPase, being the main agent for calcium uptake into the ER, plays a central role in this process. By isolation and extensive characterization of conditional mutations in the Drosophila SERCA gene, we describe novel roles of this key protein in neuromuscular physiology and enable a genetic analysis of SERCA function. At motor nerve terminals, SERCA inhibition retards calcium sequestration and reduces the amplitude of evoked excitatory junctional currents. This suggests a direct contribution of store-derived calcium in determining the quantal content of evoked release. Conditional paralysis of SERCA mutants is also marked by prolonged neural activity-driven muscle contraction, thus reflecting the phylogenetically conserved role of SERCA in terminating contraction. Further analysis of ionic currents from mutants uncovers SERCA-dependent mechanisms regulating voltage-gated calcium channels and calcium-activated potassium channels that together control muscle excitability. Finally, our identification of dominant loss-of-function mutations in SERCA indicates novel intra- and intermolecular interactions for SERCA in vivo, overlooked by current structural models.

Animals↗
PubMed2000

Behavioral transformations during metamorphosis: remodeling of neural and motor systems.

During insect metamorphosis, neural and motor systems are remodeled to accommodate behavioral transformations. Nerve and muscle cells that are required for larval behavior, such as crawling, feeding and ecdysis, must either be replaced or respecified to allow adult emergence, walking, flight, mating and egg-laying. This review describes the types of cellular changes that occur during metamorphosis, as well as recent attempts to understand how they are related to behavioral changes and how they are regulated. Within the periphery, many larval muscles degenerate at the onset of metamorphosis and are replaced by adult muscles, which are derived from myoblasts and, in some cases, remnants of the larval muscle fibers. The terminal processes of many larval motoneurons persist within the periphery and are essential for the formation of adult muscle fibers. Although most adult sensory neurons are born postembryonically, a subset of larval proprioceptive neurons persist to participate in adult behavior. Within the central nervous system, larval neurons that will no longer be necessary die and some adult interneurons are born postembryonically. By contrast, all of the adult motoneurons, as well as some interneurons and modulatory neurons, are persistent larval cells. In accordance with their new behavioral roles, these neurons undergo striking changes in dendritic morphology, intrinsic biophysical properties, and synaptic interactions.

Animals↗
PubMed2000

Remodeling of the femoral chordotonal organ during metamorphosis of the hawkmoth, Manduca sexta.

During metamorphosis of the moth, Manduca sexta, the larval legs degenerate and are replaced by adult legs with a diverse array of new sensory organs. The majority of the larval sensory neurons degenerate but some hair sensilla and chordotonal organ sensory neurons survive metamorphosis (Consoulas [2000] J. Comp. Neurol. 419:154-174). In the present study nerve-tracing techniques, birth-date labeling (5-bromodeoxyuridine), and electrophysiology were used to describe the remodeling of the femoral chordotonal organ (FCO) in the prothoracic legs. The larval FCO is composed of two scoloparia, which are associated with separate apodemes. At the onset of metamorphosis, some of the 13 larval neurons degenerate, together with the larval FCO apodemes. The remaining larval FCO sensory neurons persist in the imaginal leg to become the precursors of the adult femoral and tibial chordotonal organs respectively. Early in the pupal stage, 45 to 60 new sensory neurons are generated de novo and become associated with 6 persistent larval neurons in the imaginal femur to compose the adult FCO. Two clusters of persistent and new neurons are enclosed into two scoloparia with short apodemes that eventually become fused. In both larval and adult stages, the FCO contains units that respond phasically and others that respond tonically to femorotibial movements. Nerve activity from the FCO neurons can be recorded continuously during the remodeling of the organ. A persistent leg flexor motoneuron receives inputs from the FCO sensory neurons in both larval and adult stages, offering the opportunity to investigate the remodeling of the neural circuits underlying the proprioceptive control of the femorotibial joint.

Afferent Pathways↗
PubMed2000

Remodeling of the leg sensory system during metamorphosis of the hawkmoth, Manduca sexta.

The adult legs of the hawkmoth Manduca sexta are supplied by a diverse array of sensory organs and associated neurons (Kent and Griffin [1990] Cell Tissue Res. 259:209-223) that differ from those in the larval legs. In the present study, a combination of nerve-tracing techniques [biocytin, 1,1;-dioctadecyl-3,3,3;, 3;-tetramethyl-indocarbocyanine perchlorate (DiI)], birth date labeling (5-bromodeoxyuridine), confocal microscopy, and electrophysiology were used to describe the remodeling of the prothoracic leg sensory system. Four primary sensory branches carry the axons of all of the sensory neurons in the larval leg. At the onset of metamorphosis, the imaginal leg epidermis develops underneath the larval cuticle and encircles the sensory neurons, thus separating them from their target-organs. Most of the larval neurons degenerate during the larval-to-pupal transition and are replaced by new-adult sensory neurons that are born and differentiate in the pupa. Six sensory neurons that supply hair sensilla in the larval leg, together with 13 femoral and tibial chordotonal organ neurons, persist into the developing adult leg to serve similar functions. Early in the pupal stage, electrical activity can be recorded from these neurons despite the absence of target sensory structures. During the differentiation of the adult sensory system, the axons of the new-adult sensory neurons contact and fasciculate with the axons of the persistent neurons. Thus, five of the primary sensory branches of the adult leg are built on the preexisting larval sensory trajectories. Two sensory branches, however, are established de novo by the axons of specific adult sensory neurons.

Aging↗
PubMed1999

Peripheral distribution of presynaptic sites of abdominal motor and modulatory neurons in Manduca sexta larvae.

Insect muscle fibers are commonly innervated by multiple motor neurons and efferent unpaired median (UM) neurons. The role of UM neurons in the modulation rather than rapid activation of muscle contraction (Evans and O'Shea [1977] Nature 270:257-259) suggests that their terminal varicosities may differ structurally and functionally from the presynaptic terminals of motor neurons. Furthermore, differences in the characteristics of UM neuron terminal varicosities may be correlated with functional differences among their diverse target muscles. Larval abdominal body wall muscles in the hawkmoth, Manduca sexta, consist of large, elongated fibers that are multiterminally innervated by one and occasionally two motor neurons (Levine and Truman [1985] J. Neurosci. 5:2424-2431). The fibers are also innervated by one of two efferent UM neurons that bifurcate to innervate targets on both sides of the abdomen (Pflüger et al. [1993] J. Comp. Neurol. 335:508-522). In this study, the intracellular tracer biocytin was used to identify the targets of the UM neurons and to distinguish their terminal axonal varicosities on the muscle fibers. An antiserum to the synaptic vesicle protein, synaptotagmin, was used to label synaptic vesicles, and the styryl dye FM1-43 was used to demonstrate release and recycling. Most of the abdominal muscles in a given hemisegment were found to be supplied by one of the two UM neurons. Terminal varicosities of the excitatory motor neurons were large (3-7 pm) and were found in rows of rosettes that extended to every aspect of the muscle fiber; these varicosities were designated as type I terminals. The UM neuron terminal varicosities also occupied every aspect of the fiber but were smaller (1-3 microm) and more separated from each other; these were designated as type II terminals. Both type I and type II terminals are synaptotagmin immunoreactive and, as shown by FM1-43 staining, are sites of synaptic vesicle recycling. The excitatory motor neuron terminals (type I) could easily be loaded and unloaded with FM1-43, which indicates their capacity for repeated vesicular exocytosis and recycling. In contrast, the dye could not as readily be unloaded from UM neuron terminals (type II), which may indicate that they have a slower turnover of synaptic vesicles.

Abdominal Muscles↗
PubMed1999

Digger wasp versus cricket: immediate actions of the predator's paralytic venom on the CNS of the prey.

The females of the palaearctic digger wasp species Liris niger hunt crickets (e.g., Acheta domesticus) as food for their future brood. The wasps paralyze the prey by injecting their venom directly into each of the three thoracic ganglia and the suboesophageal ganglion. This study describes the effects produced by the Liris venom at the level of the intact prey animal (by chronic electromyogram) and at the level of a dissected preparation (by extra- and intracellular records) during the immediate action. Natural or artificial injections of the Liris venom into various ganglia revealed that: (a) The venom injection induced an about 15- to 35-s long tonical discharge of the neurons located in the stung ganglion. This discharge is usually accompanied by convulsions of the prey's limbs. (b) Subsequently, the generation and propagation of action potentials are blocked for up to 30 min (total paralysis). (c) During total paralysis, the venom blocks synaptic transmission. (d) The effects of the venom are restricted to the stung ganglion. Responses of mechanoreceptors in the legs can be recorded from the peripheral nerves of the stung ganglion during the whole period of total paralysis. (e) The neurons almost completely recover after this period. The venom does not selectively affect leg motoneurons, but affects any neuron (e.g., internerneurons or neurosecretory neurons) in any part of the central nervous system of the prey where it was released.

Action Potentials↗
PubMed1999

Patterned activation of unpaired median neurons during fictive crawling in manduca sexta larvae

The unpaired median neurons are common to the segmental ganglia of many insects. Although some of the functional consequences of their activation, among them the release of octopamine to modulate muscle contraction, have been described, less is understood about how and when these neurons are recruited during movement. The present study demonstrates that peripherally projecting unpaired median neurons in the abdominal and thoracic ganglia of the larval tobacco hornworm Manduca sexta are recruited rhythmically during the fictive crawling motor activity that is produced by the isolated central nervous system in response to pilocarpine. Regardless of the muscles to which they project, the efferent unpaired median neurons in all segmental ganglia are depolarized together during the phase of the crawling cycle when the thoracic leg levator motoneurons are active. During fictive crawling, therefore, the unpaired median neurons are not necessarily active in synchrony with the muscles to which they project. The rhythmical synaptic drive of the efferent unpaired median neurons is derived, at least in part, from a source within the subesophageal ganglion, even when the motor pattern is evoked by exposing only the more posterior ganglia to pilocarpine. In pairwise intracellular recordings from unpaired median neurons in different ganglia, prominent excitatory postsynaptic potentials, which occur with an anterior-to-posterior delay in both neurons, are seen to underlie the rhythmic depolarizations. One model consistent with these findings is that one or more neurons within the subesophageal ganglion, which project posteriorly to the segmental ganglia and ordinarily provide unpatterned synaptic inputs to all efferent unpaired median neurons, become rhythmically active during fictive crawling in response to ascending information from the segmental pattern-generating network.

Journal Article↗
PubMed1998

Presynaptic function during muscle remodeling in insect metamorphosis.

During metamorphosis the leg neuromuscular system of the moth Manduca sexta undergoes an extensive remodeling as the larval muscles degenerate and are replaced by new muscles in the adult. The terminal processes of persistent leg motoneurons undergo severe regression followed by regrowth (Consoulas et al., 1996), accompanied, as shown here, by the loss and re-establishment of functional presynaptic specializations. Before and shortly after the degeneration of the larval muscle, immunoreactivity for the vesicular protein synaptotagmin was localized to the presynaptic varicosities of the motoneurons. Similarly localized were distinct sites of Ca2+-dependent uptake of the fluorescent dye FM1-43. During myoblast migration and accumulation about the re-expanding motor axons, synaptotagmin immunoreactivity was widely distributed in axons, and specific FM1-43 staining revealed vesicle exocytosis in distal axon branches. During myoblast proliferation and fusion, and myotube formation, synaptotagmin staining remained widely distributed in nerve branches, whereas FM1-43 staining was more localized to subdomains of these nerve branches. These initial presynaptic active sites were transient and were replaced by new sites in more distal nerve processes as the muscle anlage increased in size and additional myotubes formed. After myotube separation, synaptotagmin staining disappeared from primary branches but remained distributed within secondary and high-order nerve branches. FM1-43 staining was detected in high-order branches only. During muscle fiber striation, growth, and maturation, both FM1-43 staining and synaptotagmin immunoreactivity became localized to terminal varicosities. Thus, presynaptic function can persist after the loss of the target and occurs transiently in axon shafts before becoming restricted to terminal domains as the underlying muscle fibers mature.

Animals↗
PubMed1997

Accumulation and proliferation of adult leg muscle precursors in Manduca are dependent on innervation.

During metamorphosis, the larval thoracic legs of the moth Manduca sexta are replaced by new adult legs. The leg motoneurons do not die after the loss of the larval muscles, but persist to innervate the new adult leg muscles (Kent and Levine, 1988). The adult muscles form from myoblasts that originate in specific production sites within the legs and migrate to the sites of muscle formation, where they accumulate, proliferate, and fuse to form myofibers (Consoulas et al., 1996b). Throughout adult leg muscle development, there is a close association between nerves and the developing muscles, suggesting a role for the nervous system in myogenesis (Consoulas et al., 1996a). This prediction was confirmed and the role of the nervous system clarified in the present study by cutting the larval leg nerves prior to metamorphosis. Although myoblasts were generated and migrated normally in the operated leg, they failed to accumulate in the appropriate regions. The myoblasts did not die, but failed to proliferate and remained in the denervated legs as dispersed cells or as aggregates in inappropriate regions. In about 26% of cases, this resulted in the formation of adult legs that lacked muscles. In the remaining cases, however, delayed regeneration of the leg nerve occurred and small muscles appeared in the more proximal segments of the denervated legs. Each muscle fiber in these operated legs bore motor terminals belonging to axons of the leg nerves which had grown out from the proximal nerve stump and invaded the leg. Following the delayed appearance of motor axons, myoblasts aggregated and underwent proliferation and differentiation into muscle fibers. In a second set of experiments, denervation was performed later, after myoblasts had aggregated to establish anlagen. Myoblast proliferation was reduced but differentiation continued. These observations suggest that motor nerves are essential for both the accumulation of myoblasts into the correct areas of muscle development and the appropriate level of proliferation.

Aging↗
PubMed1997

Development of adult thoracic leg muscles during metamorphosis of the hawk moth Manduca sexta.

During metamorphosis, the larval thoracic legs of the hawk moth Manduca sexta are replaced by a new set of adult legs. The larval leg motoneurons persist to innervate new adult muscles, and the motor terminals remain within the developing adult legs. Here we describe the fate of the larval leg muscles and the origin of new muscles within the adult legs. During the larval instars, large and small nuclei proliferate within leg muscle fibers. Near the end of the larval stage a subset of the small nuclei undergo a wave of proliferation, as indicated by the incorporation of 5-bromodeoxyuridine, whereas other nuclei die. However, none of the larval leg muscles fibers persist to serve as templates for adult muscle formation, and there was no evidence for persistence of larval myonuclei. Migrating myoblasts that are born within aggregate to form adult muscle anlagen at specific production sites within the developing imaginal legs. Intense nuclear proliferation occurs within the anlagen during the early pupal stage, followed by muscle fiber formation and striation. We conclude that adult leg muscles form mainly, if not exclusively, from migrating myoblasts that without the involvement of larval elements.

Animals↗
PubMed1996

Remodeling of the peripheral processes and presynaptic terminals of leg motoneurons during metamorphosis of the hawkmoth, Manduca sexta.

During metamorphosis of the hawkmoth, Manduca sexta, the muscles, cuticular structures, and most sensory neurons of the larval thoracic legs are replaced by new elements in the adult legs. The thoracic leg motoneurons, however, survive the loss of the larval muscles and persist to innervate new targets in the imaginal legs. Here we have used biocytin staining, immunocytochemistry, and confocal microscopy to follow the fates of the peripheral processes and presynaptic terminals of the leg motoneurons. Although the most distal processes of the motor nerves retract following the degeneration of larval leg muscles, the axon terminals always retain close association with the muscle remnants and the anlagen of the new adult muscles. As the imaginal muscles differentiate and enlarge, the motor terminals expand to form adult presynaptic terminals. An antibody to the presynaptic protein, synaptotagmin, revealed its localization to the terminal varicosities in both larval and adult stages but distribution within pre-terminal branches during adult development. Electrophysiological methods revealed that functional neuromuscular transmission first occurs quite early during metamorphosis, before the differentiation of contractile elements in the muscle fibers.

Animals↗