Extensor motor neurons of the crayfish abdomen

Summary The somata of five deep extensor motoneurons of the third abdominal ganglion of the crayfish(Procambarus clarkii) were located and identified. The positions of these somata within the ganglion and their distal distribution to muscles have been mapped and were constant. The soma of the extensor inhibitor was noted to touch the soma of the flexor inhibitor. Three of the excitatory neurons were clustered near their exit route.Sensory and cord routes of activation of the extensor motoneurons were also found and were constant from preparation to preparation. Sub-threshold recording showed that these motoneurons exhibited radically different types of post-synaptic response to stimuli at different sites in the nervous system. No interaction between extensor motoneurons or between the extensor and flexor motoneurons was observed.     

Cross-connections coordinate postural motoneuron activity in the crayfish abdomen

Intracellular recordings and dye injections were used to examine mutual coupling among slow abdominal postural motoneurons in the 4th abdominal ganglion in crayfish (Procambarus clarkii). Intracellular current injection into one motoneuron altered the spike firing rate of some of its synergists. Depending on the polarity of the injected current, the premotor effect on the synergists was excitatory or inhibitory. The magnitude of the effect was intensity dependent. No dye coupling was found among the motoneurons following injection of Lucifer yellow. The morphological basis of the coupling was examined by differential filling of motoneuron pairs, one with horseradish peroxidase and the other with Lucifer yellow. The stained motoneurons were simultaneously visualized under light microscopy to determine the proximity of their differently colored dendrites. It was thus possible to locate the site of the presumed monosynaptic contacts between them. Combined physiological and morphological evidence suggests that these neurons are mutually coupled, forming part of an integrative system for abdominal posture control in crayfish.     

Interactions between the tonic and cyclic postural motor programs in the crayfish abdomen

1. In the crayfish (Procambarus clarkii) abdomen, the superficial flexor and extensor muscles and the motoneurons that innervate them are employed during two completely different modes of behavior: (1) tonic postural adjustments and (2) cyclic movements associated with backwards terrestrial walking. We have tested the possibility that these two behavioral subsystems share at least some of the same tonic premotor interneurons. 2. Of the 108 tonic flexion- and extension-producing interneurons monitored during cyclic pattern generation, only 25 were recruited while 36 were inhibited. None of the recruited interneurons made a measurable contribution to the cyclic motor output. Similarly, none of the 20 inhibitory interneurons of the tonic subsystem recorded in this study was found to play a role in shaping the cyclic motor pattern. 3. Simultaneous activation of single tonic postural interneurons with the cyclic motor pattern revealed that the two behavioral subsystems interact in complex ways. Some tonic interneurons produced motor outputs that overrode the cyclic motor outputs while the motor outputs of other tonic interneurons were completely overwhelmed by the cyclic motor program. Still other tonic interneurons generated motor outputs that predominated over cyclic patterned outputs in some ganglia but were masked by the cyclic motor pattern in other ganglia. 4. Although weak interactions between the two subsystems occur at the premotor level, they have little effect on the normal generation of the cyclic pattern. Stronger interactions apparently occur at the level of the motoneurons and these interactions presumably may form the basis of switching from one behavior to the other. We conclude, therefore, that each behavioral subsystem relies upon its own unique set of premotor interneurons. Finally, those interneurons contributing to the cyclic motor pattern have not yet been identified.     

Physiological compensation in unilateral eyestalk ablated crayfish, Cherax quadricarinatus

The eyestalks of crustaceans contain neurosecretory cells involved in the regulation of molting. In crayfish, bilateral ablation results in increased molting frequency and weight gain whereas unilateral ablation typically has no effect on molting frequency and weight gain. The effects of unilateral ablation were examined in juvenile Australian freshwater crayfish, Cherax quadricarinatus. As observed for other crayfish species, molting frequency and weight gain of unilateral ablated crayfish were not significantly different from control (intact) crayfish. Survival of unilateral ablated crayfish, however, was reduced compared to controls and was likely due to stress associated with the surgical procedure itself. Using radiolabeling techniques, protein synthesis was determined for neural tissues from the remaining eyestalk of ablated crayfish and compared to protein synthesis of neural tissues from eyestalks of control, non-ablated crayfish. Protein synthesis of ablated crayfish neural tissues was significantly higher (ca. 45%) than protein synthesis of control neural tissues. Electrophoretic analysis (SDS-PAGE and autoradiography) further demonstrated that protein synthesis increased linearly for all proteins in the remaining eyestalk of ablated crayfish. Together, these results suggest that a compensatory response occurred in unilateral ablated crayfish allowing normal physiological functions, particularly those involved in regulating growth cycles, to be maintained. J. Exp. Zool. 289:184-189, 2001.     

Characterization of muscles associated with the articular membrane in the dorsal surface of the crayfish abdomen

The anatomy, physiology, and biochemistry of the dorsal membrane muscle (DMA) and the superficial extensor muscle accessory head (SEAcc) in the abdomen of the crayfish, Procambarus clarkii and lobster, Homarus americanus, are reported. These muscles have not been previously characterized physiologically or biochemically. The anatomy was originally described by Pilgrim and Wiersma (1963. J Morph 113:453-587). The arrangement of these muscles varies depending on the abdominal segment. The function of the dorsal membrane muscle is to retract the thin articulating membrane joining the cuticular segments so that the dorsal membrane does not evert during extension of the abdomen. Consequently, the articular membrane does not protrude, and thus potential damage to the membrane is minimized. Examination of nerve terminal morphology revealed strings of varicosities, usually only associated with tonic terminals. The electrophysiological data indicate that there are at least four tonic excitatory and one inhibitory motor neuron innervating these muscles. Facilitation indices and fatigue-resistance indicate physiologically the tonic nature of innervation. Anti-GABA antibodies demonstrate the anatomical presence of an inhibitor motor neuron. The SDS electrophoretic analysis of myofibrillar proteins and Western blots of key protein isoforms for these muscles in crayfish and lobsters also indicate that the DMA and SEAcc muscles are tonic phenotype. J. Exp. Zool. 287:353-377, 2000.     

High activity of NADP-dependent malic enzyme in mitochondria from abdomen muscle of the crayfish Orconectes limosus.

1. Mitochondria isolated from abdomen muscle of crayfish Orconectes limosus exhibit malic enzyme activity in the presence of L-malate, NADP and Mn2+ ions after addition of Triton X-100. Under optimal conditions about 230 nmole of reduced NADP and an equivalent amount of pyruvate are produced per min per mg of mitochondrial protein. 2. The pH optimum for decarboxylation of L-malate is about 7.5. 3. The apparent Km for L-malate, NADP and Mn2+ ions was found to be 0.66, 0.012, and 0.0025 mM, respectively. 4. The requirement for Mn2+ can be replaced by Mg2+, Co2+ and Ni2+ ions; however, higher concentrations of these ions than Mn2+ are required for a full stimulation of malic enzyme activity. 5. Oxaloacetate and pyruvate inhibited the enzyme activity in a competitive manner with apparent Ki values of 0.05 mM and 5.4 mM, respectively.     

Load compensation in obese patients during quiet tidal breathing

Seventeen eucapnic massively obese patients and eight normal subjects had their respiratory cycle parameters studied while breathing room air at rest. Despite large variations in the degree of obesity, our patients demonstrated normal mean inspiratory and expiratory flow rates, duty cycles, and minute ventilation. The maintenance of normal mean inspiratory flow rates was found to be dependent on an augmentation of neuromuscular drive (P0.1); furthermore, a strong positive correlation between percentage ideal body weight (i.e., the degree of obesity) and P0.1 was present. The obese were found to partition their tidal volume preferentially to their rib cage compartment, choosing to leave the abdominal compartment relatively immobile. Analysis of the diaphragmatic electromyogram revealed a persistence of activity into early expiration, the length of which also depended on the degree of obesity. These findings suggest that the diaphragm's volume-generating function in the obese is reduced, and furthermore the persistence of its activity in expiration serves to attenuate the rate of expiratory flow. No significant difference in any respiratory cycle parameter was found between simple obesity patients and formerly hypercapnic obese patients.     

Mitochondrial malic enzyme from the crayfish abdomen muscle,purification, regulatory properties and possible physiological role

• 1.1. Mitochondrial malic enzyme (l-Malate: NADP oxidoreductase (oxaloacetate decarboxylating) EC 1.1.1.40) has been isolated from abdomen muscle of crayfish Orconectes limosus by chromatography on Sepharose 6B and DEAE cellulose. Specific activity of the purified enzyme was about 5 μmols per min per mg protein, which corresponds to about 30-fold purification.
• 2.2. This enzyme showed extremely small reversiblity, since the reaction in the direction of decarboxylation is at least 37, 190 and 760 times that for the carboxylation at pH 7.0, 7.5 and 8.0 respectively.
• 3.3. Purified enzyme showed allosteric properties, which was more accentuated at more alkaline pH (Hill coefficients were 1.1, 1.7 and 1.8 at pH 7.0, 7.5 and 8.0 respectively). The activity of malic enzyme was increased considerably in the presence of succinate and fumarate.
• 4.4. Mitochondira isolated from abdomen muscle of Orconectes limosus incubated in the presence of malate, fumate and succinate catalysed pyruvate production which was stimulated by ADP and inhibited by respiratory chain inhibitors.
• 5.5. NADH but not NADPH oxidation was catalysed by broken mitochondria or sonic particles. When NADPH and NAD were added simultaneously the rate of oxidation. This suggests the presence of active NADPH:NAD transhydrogenase in mitochondria isolated from the crayfish abdomen muscle.
• 6.6. A possible metabolic role for NADP-linked malic enzyme/transhydrogenase couple in abdomen muscle of crayfish Orconectes limosus is proposed.

     

Motor pattern changes during central compensation of eyestalk posture after unilateral statolith removal in crayfish

Electromyographic recording was used to study how the activity of the eyestalk motor system is modified during the recovery of eyestalk posture following unilateral statolith removal in crayfish Procambarus clarkii Girard. Intact animals showed bilaterally balanced activity of the muscle 12 (eyecup-up muscle) in the upright body position. Body rolling caused an increase in the muscle activity on the lowered side and a decrease on the lifted side. Unilateral statolith removal caused imbalance in the bilateral muscle activity in the upright body position: the muscle 12 activity decreased tonically on the operated side and increased on the opposite side. Body rolling of the operated animal caused an increase in the muscle activity from the unbalanced level on the lowered side and a decrease on the lifted side. When the operated animal recovered its original symmetrical posture of eyestalks 14 days after operation, the muscle activity was found on both sides to return to the previous level observed before statolith removal, regardless of the post-operative condition in which the animal was maintained. In those animals that did not recover the original eyestalk posture, the unbalanced activity of bilateral muscles that was caused by unilateral statolith removal remained unchanged. The results indicate that the recovery of eyestalk posture is based on restoration of the original activity balance, rather than on fixation of the operation-induced activity imbalance, among bilaterally homologous sets of muscles in the course of central compensation.     

Load compensation and respiratory muscle function during sleep.

The sleeping state places unique demands on the ventilatory control system. The sleep-induced increase in airway resistance, the loss of consciousness, and the need to maintain the sleeping state without frequent arousals require the presence of complex compensatory mechanisms. The increase in upper airway resistance during sleep represents the major effect of sleep on ventilatory control. This occurs because of a loss of muscle activity, which narrows the airway and also makes it more susceptible to collapse in response to the intraluminal pressure generated by other inspiratory muscles. The magnitude and timing of the drive to upper airway vs. other inspiratory pump muscles determine the level of resistance and can lead to inspiratory flow limitation and complete upper airway occlusion. The fall in ventilation with this mechanical load is not prevented, as it is in the awake state, because of the absence of immediate compensatory responses during sleep. However, during sleep, compensatory mechanisms are activated that tend to return ventilation toward control levels if the load is maintained. Upper airway protective reflexes, intrinsic properties of the chest wall, muscle length-compensating reflexes, and most importantly chemoresponsiveness of both upper airway and inspiratory pump muscles are all present during sleep to minimize the adverse effect of loading on ventilation. In non-rapid-eye-movement sleep, the high mechanical impedance combined with incomplete load compensation causes an increase in arterial PCO2 and augmented respiratory muscle activity. Phasic rapid-eye-movement sleep, however, interferes further with effective load compensation, primarily by its selective inhibitory effects on the phasic activation of postural muscles of the chest wall. The level and pattern of ventilation during sleep in health and disease states represent a compromise toward the ideal goal, which is to achieve maximum load compensation and meet the demand for chemical homeostasis while maintaining sleep state.     

Physiological changes of premotor nonspiking interneurons in the central compensation of eyestalk posture following unilateral sensory ablation in crayfish

We investigated how the physiological characteristics and synaptic activities of nonspiking giant interneurons (NGIs), which integrate sensory inputs in the brain and send synaptic outputs to oculomotor neurons innervating eyestalk muscles, changed after unilateral ablation of the statocyst in order to clarify neuronal mechanisms underlying the central compensation process in crayfish. The input resistance and membrane time constant in recovered animals that restored the original symmetrical eyestalk posture 2 weeks after operation were significantly greater than those immediately after operation on the operated side whereas in non-recovered animals only the membrane time constant showed a significant increase. On the intact side, both recovered and non-recovered animals showed no difference. The frequency of synaptic activity showed a complex pattern of change on both sides depending on the polarity of the synaptic potential. The synaptic activity returned to the bilaterally symmetrical level in recovered animals while bilateral asymmetry remained in non-recovered ones. These results suggest that the central compensation of eyestalk posture following unilateral impairment of the statocyst is subserved by not only changes in the physiological characteristics of the NGI membrane but also the activity of neuronal circuits presynaptic to NGIs.     

Load compensation as a function of state during sleep onset

Ventilation decreases and airway resistanceincreases with the loss of electroencephalogram alpha activity at sleeponset. The aim of this study was to determine whether reflexive load compensation is lost immediately on the loss of alpha activity. Sixhealthy male subjects were studied under two conditions (load andcontrol-no load), in three states (continuous alpha, continuous theta,and immediately after a transition from alpha to theta), and in twophases (early and late sleep onset). Ventilation and respiratory timingwere measured. A comparison of loaded with control conditions indicatedthat loading had no effect on inspiratory minute ventilation duringcontinuous alpha (differential effect of 0.00 l/min) and only a small,nonsignificant effect in theta immediately after phase2 transitions (0.31 l/min), indicating a preservationof load compensation at these times. However, there were significantdecreases in inspiratory minute ventilation on loaded trials duringcontinuous theta in phase 2 (0.77 l/min) and phase 3 (1.15 l/min) andduring theta immediately after a transition in phase3 (0.87 l/min), indicating a lack of reflexive loadcompensation. The results indicate that, because reflex load compensation is state dependent, state-related changes in airway resistance contribute to state-related changes in ventilation duringsleep onset. However, this effect was slightly delayed with transitionsinto theta early in sleep.

     

Muscle receptor organs do not mediate load compensation during body roll and defense response extensions in the crayfish Cherax destructor.

It has been proposed that the abdominal muscle receptor organ (MRO) of decapod crustaceans acts in a sensory feedback loop to compensate for external load. There is not yet unequivocal evidence of MRO activity during slow abdominal extension in intact animals, however. This raises the possibility that MRO involvement in load compensation is context-dependent. We recorded from MRO tonic stretch receptors (SRs) in freely behaving crayfish (Cherax destructor) during abdominal extension occurring during two different behaviors: body roll and the defense response. Abdominal extensions are similar in many respects in both behaviors, although defense response extensions are more rapid. In both situations, SR activity typically ceased when the abdominal extension commenced, even if the joint of the SR being monitored was mechanically prevented from extending by a block. Since extensor motor neuron activity increased when the abdomen was prevented from extending, we concluded that the load compensation occurring in these behaviors was not mediated by the MROs.